Management ofAbstract Acute Gouty Arthritis with …referring to podagra as‘arthritis of the...
Transcript of Management ofAbstract Acute Gouty Arthritis with …referring to podagra as‘arthritis of the...
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1Hippocratic Journal of Unani Medicine
GAbstract
out is one of the most common types of inflammatory joint
diseases; affects an estimated 1-1.5% of the world population. Modern drugs
used for subsiding acute attacks or lowering serum uric acid are associated
with potent adverse effects. Moreover, these commonly used therapeutic agents
often, and for various reasons, do not achieve the desired lowering of serum
urate levels to below 6.0 mg/dl. On the basis of conventional Unani Usool-e-
Ilaj of Niqris (gout), five herbal drugs from the list of classical Unani anti-
arthritic drugs have been selected and formulated in capsule form and a single
blind placebo controlled clinical trial was carried out to evaluate the efficacy
and safety of this capsule in the management of gouty arthritis. Six week
treatment with test drug produced remarkable effects on various efficacy
parameters. No any adverse effect was observed during the course of treatment.
Keywords: Gout, Niqris, Wajaul Mafasil, Suranjan, Cocchicum luteum
Introduction
Gouty arthritis is among the earliest diseases that have been recognized as
a clinical entity. First identified by the Egyptians in 2640 BC, podagra (acute
gout occurring in the first metatarsophalangeal joint) was later recognized by
Buqrat in the fifth century BC, who referred to it as ‘the unwalkable disease.
Buqrat also noted the link between the disease and an intemperate lifestyle,
referring to podagra as‘arthritis of the rich’, as opposed to rheumatism, an
‘arthritis of the poor’(Nuki et al., 2006). Six centuries later to Buqrat, Jalinoos
was the first to describe tophi (James et al., 2000).
Gout is one of the most common types of inflammatory joint diseases; affects
an estimated 1-1.5% of the world population (Praveen et al., 1994). The
prevalence of gout is rising as a result of a changing pattern of lifestyle
(Arromdee et al., 2002). In most cases, no identifiable underlying cause of
gout is present, but evident factors are usually present that could contribute
to increase in urate (uric acid) levels, such as renal function disorders, obesity,
and the use of thiazide diuretics (Roubenoll, 1990). Although, hyperuricemia is
a risk factor for the development of gout, the exact relationship between
hyperuricemia and acute gout is unclear. Acute gouty arthritis can occur in the
presence of normal serum uric acid concentrations. Conversely, many persons
with hyperuricemia never experience an attack of gouty arthritis (McCarty,
1994).
Management ofAcute GoutyArthritis with aPolyherbalUnaniFormulation
1Rais ur Rahman,2Dania Siddiqui,
2Naseem Akhtar,2D.S. Dua
and3*Yasmeen Shamsi
1Department of AYUSH
Ministry of Health & F.W.,
Government of India,
GPO Complex, INA,
New Delhi - 110023
2Department of Moalijat
Ayurvedic and Unani Tibbia College,
Karol Bagh, New Delhi-11005
3Department of Mahiyatul Amraz
Faculty of Medicine (U),
Jamia Hamdard, New Delhi-110062
July - September 2014, Vol. 9 No. 3, Pages 1-13
3*Author for correspondence
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2Hippocratic Journal of Unani Medicine
Several approaches to the treatment of gout are available depending on the
patient’s presentation of the disease, patient’s specific risk factors (high serum
urates, previous attacks and radiographic signs), the clinical phase of the
disease (acute, recurrent, tophaceous) and general risk factors, such as obesity
and alcohol consumption.
Acute gout is usually treated by reducing inflammation of the affected joint with
non-steroidal anti-inflammatory drugs (NSAIDs), colchicine, corticosteroids etc.
Although these agents are generally effective, they also present significant
risks in patients who have pre-existing renal, cardiovascular, metabolic and
gastrointestinal diseases (Nuki, 1999; Emmerson, 1996).
Antihyperuricemic drugs such as allopurinol, benzbromarone, sulfinpyrazone
and probenecid can have potent side effects (Singer et al., 1986, Arellano et
al., 1993). Benzbromarone was withdrawn from the market in Europe in 2003,
but was registered again in some countries in 2004 (Sutaria et al., 2006). Its
use is now restricted for patients with gout who are allergic to allopurinol or
those in whom allopurinol is contraindicated (Jansen et al., 2004). Furthermore,
these commonly used therapeutic agents often, and for various reasons, do
not achieve the desired lowering of serum urate levels to below 6.0 mg/dl.
The side effects/drawbacks of all above mentioned drugs call for the development
of novel drugs with similar or better efficacy and lesser toxicity than presently
available drugs.
On the basis of conventional Unani Usool-e-Ilaj of Niqris (gout), five herbal
drugs from the list of classical Unani anti-arthritic drugs have been selected
and formulated in capsule form and a clinical trial was carried out to evaluate
the efficacy and safety of this capsule in the management of gout.
Materials and Methods
Study Drug
Study drug was a combination of five herbs namely, Suranjan (Colchicum
luteum), Elva (Aloe barbadensis), Qurtum (Carthamus tinctorius), Halaila-e-
Zard (Terminalia chebula), Zanjbeel (Zingiber officinale). All these five drugs in
equal proportion were finely powdered and encapsulated in hard gelatin capsule
in the quantity of 1gm.
Placebo
Placebo was supplied to the patients in the form of similar capsules of 1gm
each containing wheat flour.
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3Hippocratic Journal of Unani Medicine
Study Design
This was a randomized, single blind, placebo controlled study, conducted in
the Department of Moalejat, A & U Tibbia College & Hospital, Karol Bagh, New
Delhi, From September 2009 to December 2011.
Participants
(i) Inclusion Criteria
Both male and female patients aged between 18 -65 years fulfilling the criteria
of American College of Rheumatology (ACR) for diagnosing acute gouty arthritis
including clinical features, laboratory and radiographic findings were included
in the study, who had serum uric acid level more than the upper limit of normal
range ( > 7 mg/dl)
(ii) Exclusion Criteria
Patients were excluded if they had renal or hepatic insufficiency or cardiovascular
disorders. Patients taking thiazide group of diuretics/ aspirin/NSAIDs and
Pregnant and lactating women were too excluded from the study.
Ethical Consideration
All patients were included in the study after obtaining written informed consent
and study was conducted according to Good Clinical Practice guidelines.
Dosage and Administration
Following 5-7 days washout period of anti-inflammatory/ analgesic drugs (e.g.,
NSAIDs, Corticosteroids) or antihyperuricemic drugs (eg allopurinol) or any
other medication used for the treatment of arthritis (e.g. Ayurvedic, Homeopathic
or Unani drugs), patients were randomly assigned to receive either drug or
placebo capsule in the dose of 2 capsule thrice daily with plain water up to a
period of six weeks. Randomization was done by lottery method.
Follow up and Drug Compliance
Clinical as well as laboratory evaluation was performed and recorded at the
baseline, week 1, week 4 and week 6. Compliance with treatment drug/placebo
was evaluated at each follow up visit by capsule count.
Criteria for the Assessment of Efficacy
To assess the response of treatment on patients of gouty arthritis in both
groups, the following parameters were used.
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4Hippocratic Journal of Unani Medicine
Subjective Parameters
• Pain (Wong-Baker’s Faces rating scale; with 0=doesn’t hurt, 2=hurts a
little bit, 4=hurts a little more, 6=hurts even more, 8=hurts a lot, 10=as
much as the patient can imagine), (Cheng et al., 2004; Taylor et al.,
2007).
• Tenderness (0-4 point scale; with 0=no tenderness, 1=patient says it is
painful, 2=patient says it is painful, winces, and pulls back, 4=patient
does not allow palpation), (Cheng et al., 2004; Taylor et al., 2007).
• Joint swelling (0-4 point scale; with 0=no swelling,1=barely perceptible,
2=mild, 3=moderate, 4=severe, bulging beyond the joint margins) (Cheng
et al., 2004; Taylor et al., 2007).
• Movement/Mobility (0-4 point scale; with 0=full voluntary
movement,1=partial voluntary movement, 2=full movement when the
joint is moved by the examiner, 3=partial movement when the joint is
moved by the examiner, 4=no movement at all)
• Serum uric acid, C-Reactive proteins, Erythrocyte Sedimentation Rate
and total leukocyte count
Assessment of Safety
To establish the safety of test drug , the following investigations were carried
out at baseline, after one week and just after the termination of treatment.
� Liver function test –S.Bilirubin, S.G.O.T, S.G.P.T. & S.Alkaline
Phosphatase
� Kidney function test – Blood Urea, S. Creatinine.
� Haemogramme – Hb%, TLC, DLC, E.S.R.
Statistical Analysis
The differences in pretreatment and post treatment obtained in both groups
(test group and control group) were compared by applying Mann-Whitney U
Test.
Results
Total 46 cases, 21 in test group and 25 in control group were randomly
enrolled in the study. Two subjects from test group and 4 patients from control
(placebo) group dropped out of the study due to unknown reason. Twenty
patients in test group and 20 in control group completed the treatment up to
the end of study (6 weeks).The demographic data and other characteristics at
baseline are presented in Table 1.
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5Hippocratic Journal of Unani Medicine
Table 1: Demographic Data Baseline Characteristics of Study Patients
Variable Test Group (N=20) Control Group (N=20)
Age (Years) Mean ± SD 42.70 ± 11.15 43.80± 9.28
Gender Male- N (%) 14 (70%) 15(75%)
Female-N (%) 06 (30%) 05 (25%)
Family H/o Gout-N (%) 2 (10%) 3 (15%)
Pain Score (Mean ± SD) 5.95 ± 1.54 5.95 ± 1.50
Tenderness (Mean ± SD) 2 ± 1.12 1.60 ± 1.14
Swelling (Mean ± SD) 2.20 ± 0.95 1.55 ± 0.94
Movement (Mean ± SD) 1.70 ± 0.86 1.40 ± 0.59
S. Uric Acid 8.04 ± 1.98 7.81 ± 0.91
C.R.P. 3.92 ± 2.35 4.37 ± 2.18
T.L.C. 9300 ± 1554.6 8785 ± 1786.3
E.S.R. 26.65 ± 10.49 28.70 ± 11.3
SD = standard deviation; ESR = erythrocyte sedimentation rate;
CRP = C-reactive proteins; TLC= total leukocyte count
The effects of 6 weeks treatment with test drug and control on various clinical
and laboratory parameters are described below:
Clincal Findings
Joint Pain
In test group, the mean score of pain (± SD) at baseline was 5.95 ± 1.54,
which was reduced to 4.50 ± 1.91 on the 7th day, 3.30 ± 1.72 on 28th day and
1.85 ± 1.98 on the termination of treatment (42nd day). While in control group,
the mean pain score was 5.95 ± 1.50 at baseline, which gradually increased
to 6.05 ± 1.47 on 7th day, 6.0 ± 1.07 on 28th day, and 6.35 ± 1.23 at the end
of treatment. On applying Mann- Whitney test, extremely significant difference
between the two groups was detected on 28th day (p<0.0001) and 42nd day
(p<0.0001), (Table-2).
Joint Tenderness
In test group the baseline tenderness score (mean ± SD) of 2.0 ± 1.12
decreased to 1.75 ± 0.97 (7th day), 1.05 ± 0.94 (28th day) and 0.60 ± 0.59
(42nd day). Whereas, there was very gradual and insignificant decrease in
mean tenderness score in control group, from baseline (1.60 ± 1,14) to 1.50
± 1 on 7th day, 1.55 ± 0.99 on 28th day, 1.50 ± 0.21 on termination of
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6Hippocratic Journal of Unani Medicine
Table 2: Effect of Test Drug and Control on Pain
Pain Control Group (N=20) Test Group (N=20)
0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 5.95 6.05 6.0 6.35 5.95 4.50 3.30 1.85
S.D. (±) 1.50 1.47 1.08 1.23 1.54 1.90 1.72 1.99
S.E.M. (±) 0.34 0.33 0.24 0.28 0.34 0.43 0.38 0.44
% change - 1.68 0.84 6.72 - 24.36 46.9 68.91
P value at 7th day (T/C) <0.01 (LS) (Mann-Whitney Test)
P value at P value 28th day (T/C) <0.0001 (ES) (Mann-Whitney Test)
P value 42nd day (T/C) <0.0001(ES) (Mann-Whitney Test)
SD = Standard Deviation,; S.E.M. Standard Error of Mean; LS = Less Significant,
HS = Highly Significant
treatment. The difference in tenderness in between the groups as analysed by
Mann-Whitney Test was not significant at 28th day (p>0.05) and moderately
significant at 42nd day (p<0.01), (Table-3).
Joint Swelling
In Placebo group, the mean score of swelling increased from 1.55 ± 0.94
(baseline) to 1.70 ± 0.98 on 7th day, 1.70 ± 0.98 on 28th day and1.75 ± 1.02
on 42nd day. On the other hand the mean score of swelling at baseline in test
group was 2.20 ± 0.95, 1.80 ± 0.83 on 7th day, 1.15 ± 0.67 on 28th day, and
0.45 ± 0.51 on 42nd day. The difference in between the groups as analysed
statistically was found to be non-significant (p>0.05) on 7th day & 28th day, but
extremely significant on 42nd day (p<0.001), (Table-4).
Table 3: Effect of Test Drug and Control on Tenderness
Tenderness Control Group (N=20) Test Group (N=20)
0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 1.60 1.50 1.55 1.50 2 1.75 1.05 0.60
S.D. (±) 1.14 1.0 0.99 0.94 1.12 0.97 0.94 0.59
S.E.M. (±) 0.25 0.22 0.22 0.21 0.25 0.22 0.21 0.13
% change - 6.25 3.12 6.25 - 12.5 47.5 70
P value at 7th day (T/C) 0.48 (NS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.12 (NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.003 (MS) (Mann-Whitney Test)
SD = Standard Deviation; S.E.M. Standard Error of Mean; LS = Less Significant,
NS Not Significant; MS = Moderately Significant
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7Hippocratic Journal of Unani Medicine
Restriction of Movements
In Control group, the mean score (± SD) of restriction in movements in control
group was 1.40 ± 0.59 at baseline, 1.45 ± 0.60 at 7th day, 1.45 ± 0.60 at 28th
day, 1.65 ± 0.74 at 42nd day. On the other hand, in test group the restriction
of movement was 1.70 ± 0.86 (baseline), 1.35 ± 0.59(7th day), 0.85 ± 0.67
(28th day), and 0.55 ± 0.51 (42nd day).According to Mann-Whitney test, the
difference in between the groups was moderately significant (p<0.01) on 28th
day and extremely significant (p<0.001) on 42nd day (Table-5).
Table 4: Effect of Test Drug and Control on Swelling
Joint Control Group (N=20) Test Group (N=20)
Swelling 0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 1.55 1.55 1.70 1.75 2.20 1.80 1.15 0.45
S.D. (±) 0.94 0.94 0.98 1.02 0.95 0.83 0.67 0.51
S.E.M. (±) 0.21 0.21 0.22 0.23 0.21 0.18 0.15 0.11
% change - 0 9.68 12.9 - 18.18 47.73 79.54
P value at 7th day (T/C) 0.44 (NS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.05(NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.0002 (ES) (Mann-Whitney Test)
SD = Standard Deviation; S.E.M. = Standard Error of Mean; NS = Not Significant;
ES = Extremely Significant
Table 5: Effect of Test Drug and Control on Restriction of Movements
Restriction Control Group (N=20) Test Group (N=20)
of
Movement 0 day 7th day 28th day 42nd day 0day 7th day 28th day 42nd day
Mean 1.40 1.45 1.45 1.65 1.70 1.35 0.85 0.55
S.D. (±) 0.59 0.60 0.60 0.74 0.86 0.59 0.67 0.51
S.E.M 0.13 0.13 0.13 0.17 0.19 0.13 0.15 0.11
% change – 3.57 3.57 17.86 – 20.59 50 67.65
P value at 7th day (T/C) 0.61 (NS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.008(NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.0001 (ES) (Mann-Whitney Test)
SD = Standard Deviation; S.E.M. = Standard Error of Mean;, NS = Not Significant;
ES = Extremely Significant
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8Hippocratic Journal of Unani Medicine
Laboratory Findings
Serum Uric Acid
In control group, the mean values of serum uric acid were 7.81± 0.91 at
baseline, 7.62 ± 1.13 mg/dl, 7.62 ± 1.14 mg/dl, 7.71 ± 1.82 mg/dl, on day 7th,
28th and 42nd respectively. On the contrary, in test group the change in the
serum levels of uric acid decreased from baseline of 8.04 ± 1.98 mg/dl to 6.21
± 1.62 mg/dl on 7th day, 6.27 ± 1.52 mg/dl on day 28 and 5.55 ± 1.57 mg/dl
on day 42. The difference in mean values between the two groups on all the
three follow ups was extremely significant (p<0.001) as analysed by using
Mann Whitney U test (Table-6).
C - Reactive Protein
The C-Reactive protein values observed in test group were 3.92 ± 2.35 mg/
dl at baseline 3.91 ± 2.17 mg/dl on 7th day, 2.99 ± 1.86 mg/dl on 28th day and
2.30 ± 1.42 mg/dl on 42nd Whereas, in control group the mean values of CRP
were recorded as 4.37 ± 2.18 mg/dl at baseline, 4.30 ± 2.20 mg/dl on 7th day,
3.95 ± 2.14 mg/dl on 28th day and 3.70 ± 2.03 mg/dl on 42nd day. The inter-
group difference was not significant (p>0.05) on day 7 and day 28, but was
less significant (p<0.05) on day 42 (Table-7).
Total Leucocyte Count (T.L.C.)
In Control group, the mean TLC count in control group was 8080 ± 1395/mm3
at baseline, 9105 ± 1325 /mm3 on day 7th, 9180 ± 1405 /mm3 on day 28th, and
8950 ± 1325/ mm3 on day 42nd. While in test group it was recorded as 9300
± 1554 mm3/ 8080 ± 1395/ mm3 on day 7th, 8380 ± 1505/ mm3 on day 28th,
and 7860 ± 1622/ mm3 on day 42nd. The difference in between the group was
Table 6: Effect of Test Drug and Control on Serum Uric Acid
S. Uric Control Group (N= 20) Test Group (N=20)
Acid (mg/dl) 0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 7.81 7.62 7.62 7.71 8.04 6.21 6.27 5.55
S.D.(±) 0.91 1.13 1.14 1.18 1.98 1.62 1.52 1.57
S.E.M(±) 0.20 0.25 0.25 0.26 0.44 0.36 0.34 0.35
% change 2.43 2.43 1.28 22.76 22.01 30.97
P value at 7th day (T/C) 0.0008 (ES) (Mann-Whitney Test)
P value at 28th day (T/C) 0.0007 (ES) (Mann-Whitney Test)
P value at 42nd day (T/C) <0.0001 (ES) (Mann-Whitney Test)
SD = Standard Deviation; S.E.M. = Standard Error of Mean; ES = Extremely Significant
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9Hippocratic Journal of Unani Medicine
less significant (p<0.05) at day 7th, not significant (p<0.05) at day 28th,and
again less significant(p<0.05) at day 42nd (Table-8).
Erythrocyte Sedimentation Rate (E.S.R.)
In control group, E.S.R noted was 28.70 ± 11.33/1 hr at baseline, 29.45 ±
11.07/1 hr at 7th day, 29.75 ± 10.42/1 hr at day 28th and 30.10 ± 11.19/1 hr
at 42nd day. On the other hand the E.S.R observed in test group was 25 ±
11.53/1 hr at 7th day, 25.30 ± 10.27/1 hr at 28th day, 22.30 ± 12.02/1 hr on
42nd day. The difference in ESR values between the two groups were a not
significant (p>0.05) on 7th and 28th day, whereas mildly significant difference
was observed (p<0.05) on 42nd day, (Table 9).
Table 7: Effect of Test Drug and Control on C-Reactive Proteins
C-reactive Control Group (N= 20) Test Group (N=20)
protein
(mg/dl) 0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 4.37 4.3 3.95 3.70 3.92 3.91 2.99 2.30
S.D.(±) 2.18 2.20 2.14 2.03 2.35 2.17 1.86 1.42
S.E.M(±) 0.48 0.49 0.48 0.45 0.52 0.49 0.42 0.32
% change 1.6 9.61 15.33 0.25 23.72 41.32
P value at 7th day (T/C) 0.33 (NS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.06 (NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.03 (LS (Mann-Whitney Test)
SD= Standard Deviation; S.E.M..=Standard Error of Mean;, NS= Not Significant; LS=
Less Significant
Table 8: Effect of Test Drug and Control on Total Leukocyte Count
T.L.C. Control Group (N=20) Test Group (N=20)
(cu.mm) 0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 8785 8080 8380 7860 9300 9105 9180 8950
S.D. (±) 1786.3 1395.7 1505.3 1622 1554.6 1325.3 1405.5 1325.7
S.E.M. (±) 399.43 312.09 336.59 362.69 347.62 296.34 314.27 296.43
% change - 8.02 4.61 10.53 - 2.09 1.29 3.76
P value at 7th day (T/C) 0.01 (LS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.08 (NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.01 (LS) (Mann-Whitney Test)
SD = Standard Deviation; S.E.M. = Standard Error of Mean; NS = Not Significant;
LS = Less Significant
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10Hippocratic Journal of Unani Medicine
Safety
During the course of the study, no adverse events were reported by the
patients or clinically detected by the investigator. No any significant change
from base line was observed in haemoglobin, SGOT, SGPT, S.Bilirubin, B.Urea
and S. Creatinine values in both the groups. The test formulation as well as
placebo was found well tolerated as indicated by 85% drug compliance
Discussion
Gout is one of the most common types of inflammatory joint diseases, modern
drugs used for subsiding acute attacks or lowering serum uric acid are
associated with potent adverse effects. Furthermore, these commonly used
therapeutic agents often, and for various reasons, do not achieve the desired
lowering of serum urate levels to below 6.0 mg/dl. In the present study, the
unani formulation consisting of Suranjan (Colchicum luteum), Elva (Aloe
barbadensis), Qurtum (Carthamus tinctorius), Halaila-e-Zard (Terminalia
chebula), Zanjbeel (Zingiber officinale) not only relieved various signs and
symptoms of gouty arthritis but also exerted remarkable effects on lowering
serum uric acid level and various other inflammatory markers. The relief in joint
pain, tenderness can be attributed to the analgesic activity of
Suranjan (Colchicum luteum) (Hakeem, 1991; Rainsford, 1999), Elva (Aloe
barbadensis) (Sharma et al., 2002; Sheshadri, 1976) and Zanjbeel (Zingiber
officinale) (Khare, 2004; Sharma et al, 2002). Anti-inflammatory activity of
Qurtum (Carthamus tinctorius) ( Linda, 2001), Halaila-e-Zard (Terminalia
chebula) (Sharma et al., 2002), Zanjbeel (Zingiber officinale) (Linda, 2001;
Sharma et al., 2002; Wagner et al.,1985 ) can also be considered in decreasing
Table 9: Effect of Test Drug and Control on ESR
E.S.R. Control Group (N=20) Test Group (N=20)
(mm/1sthr) 0 day 7th day 28th day 42nd day 0 day 7th day 28th day 42nd day
Mean 28.70 29.45 29.75 30.10 26.65 25.00 25.30 22.30
S.D. (±) 11.33 11.08 10.42 11.19 10.49 11.53 10.27 12.02
S.E.M. (±) 2.533 2.48 2.33 2.5 2.35 2.58 2.29 2.69
% change - 2.61 3.65 4.88 - 6.19 5.06 16.32
P value at 7th day (T/C) 0.29 (NS) (Mann-Whitney Test)
P value at 28th day (T/C) 0.27 (NS) (Mann-Whitney Test)
P value at 42nd day (T/C) 0.02 (LS) (Mann-Whitney Test
SD = Standard Deviation; S.E.M. = Standard Error of Mean; NS = Not Significant;
LS = Less Significant
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11Hippocratic Journal of Unani Medicine
pain, swelling and tenderness. Acute attack of gouty arthritis is initiated by the
precipitation of urate crystals in the synovial fluid resulting in inflammatory
response and the acute inflammatory cells (nutrophils) phagocytose urate
crystals and release a glycoprotein which further aggravates inflammation.
Colchicine, an active constituent of Suranjan (Colchicum luteum), has been
proved inhibitory to the glycoprotein released by the nutrophils in acute gouty
inflammation. Colchicine by binding with fibrillar protein tubulin has been found
to inhibit nutrophil migration in the inflammed joint. All this explains relief in
pain, swelling and tenderness because of Suranjan (Colchicum luteum) (Jean
Brunneton, 1995; Robert et al., 1983). The diuretic activity of Qurtum (Carthamus
tinctorius) (Kritikar & Basu, 1984) and laxative action of Halaila-e-Zard (Terminalia
chebula) (Barthakur et al., 1991) and Elva (Aloe barbadensis) (Blumenthal et
al., 1998) might also be helpful in reducing monosodium urate precipitation by
increased excretion of uric acid through urine and faeces respectively.
In the light of above discussion and on the basis of observations and results
obtained in this study, large scale, standard control, double blind randomized
clinical is warranted to further support the efficacy and tolerability of test
formulation in the treatment of gouty arthritis.
Conclusion
Unani formulation produced remarkable effects on various efficacy parameters
in cases of acute gouty arthritis. The drug was found safe and well tolerated,
as no adverse events were reported by the patients or clinically detected by
the investigator during the course of six week therapy.
References
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12Hippocratic Journal of Unani Medicine
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15Hippocratic Journal of Unani Medicine
IAbstract
nfertility is a common condition with important psychological,
economical, demographical and medical implications. WHO estimates that “60-
80 million couples worldwide are currently suffering from infertility.” Quilatte
Haiwane Manwiya (oligospermia) is one of the most common causes of male
infertility. In conventional system, various drugs are being used to increase
sperm count, but these drugs are expensive and their long term use produces
several adverse effects too. Therefore, keeping in the mind about the need of
the hour an open, uncontrolled clinical trial was carried out on 30 infertile
patients of Quilatte Haiwane Manwiya. Each patient was given trial formulation
Sufoofe Muallif - 5 grams once daily for 60 days. The assessment of efficacy
of treatment was carried out on the basis of objective parameters (Semen
analysis for sperm count, motility, morphology and hormone profile for serum
testosterone, LH, FSH). The results were analyzed statistically by using Wilcoxon
matched-paired signed ranks test, one tail paired-T test and Kruskal- Wallis
test. After the trial, significant improvement was found in various objective
parameters; sperm count (10.53 ± 1.30 to 18.06 ± 2.57, P<0.001), sperm
motility (21.70 ± 3.80 to 46.56 ± 5.06, P<0.001) and sperm morphology (76.2
± 6.40 to 91.9 ± 4.22, P<0.01). The changes in hormone profile (serum
Testosterone, LH, FSH) were not significant (P>0.05). Safety parameters were
remained within normal limits after the trial. This study suggests that Sufoof
Muallif has exhibited a good response in the improvement of semen markers;
sperm count, sperm motility and sperm morphology. Furthermore, no adverse
effect was observed and safety parameters remained within normal limits. Thus
it might be concluded that the Test drug is safe and effective for the treatment
of Quilatte Haiwane Manwiya (oligospermia).
Keywords: Quilatte Haiwane Manwiya, Oligospermia, Unani medicine, Sufoofe
Muallif, Quillate mani
Introduction
Infertility is defined as the failure of a couple to achieve conception after one
year of regular unprotected intercourse (Bradon et al., 2002; Jonathan, 2002).
Infertility is a distressing problem for about 10-15% of world’s population, with
the incidence increasing over the years (Leon Speroff et al., 1999). However,
it affects both men and women; male factor contributes for about 30-40%
cases of infertility (Sengupta et al., 1998). A recent study has indicated that
there is a decrease in sperm density over a period of past fifty years (Dhaliwal
et al., 2001). Despite of remarkable advancement in pharmacotherapy, infertility
ClinicalEvaluation ofEffect ofSufoofe Muallifin theManagement ofQuilatteHaiwaneManwiya(Oligospermia)
1*Shaikh Imtiyaz,2Mohd. Anwar
and3Mohd. Nayab
1&3Department of Ilaj bit Tadbeer,
National Institute of Unani Medicine,
Kottigepalaya, Magadi Main Road,
Bangalore-560091
2Department of Ilaj bit Tadbeer,
Ajmal Khan Tibbia College,
Aligarh Muslim University,
Aligarh-202002
July - September 2014, Vol. 9 No. 3, Pages 15-24
1*Author for correspondence
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16Hippocratic Journal of Unani Medicine
continues to raise significant economic and personal burden to the society.
Male infertility, with its clinical and psychosocial implications, poses a significant
challenge to the physicians and to the society as a whole.
Quilatte Haiwane Manwiya (oligospermia) is one of the most common causes
of male infertility. It is a condition in which the sperm count is decreased to less
than 20 million/ml of semen (Walsh, 2002). The important causes include
varicocoele, cryptorchidism, klinefelter’s syndrome, damaged testes due to
trauma or infections (tuberculosis, syphilis, mumps), neoplasm of testes, kidney
and liver diseases, smoking, alcoholism, wearing tight under garments, working
at high temperature places like welding, dyeing, blast furnace, cement and
steel factories etc (Smith, 1997).
In conventional system, various drugs like testosterone, clomiphene citrate,
tamoxifene and HCG are being used to increase sperm count, but these drugs
are expensive and their long term use produces several adverse effects like
gynaecomastia, hepatic carcinoma, deep vein thrombosis, lowering of HDL and
rise in LDL etc (Tripathi, 2006). However, in Unani system of medicine there
are ample of single as well as compound drugs which are used for male sexual
weakness (Zoafe bah) and infertility since the time immemorial.
In Unani system of medicine, most of Unani scholars particularly Ibn Sina (980-
1037 AD), Zakaria Razi (865-925 AD), Ismail Jurjani (1110 AD) and Rabban
Tabri (810-895 AD) have elaborately discussed sexual diseases in their
respective treatises. They have mentioned the causes, symptoms, complications,
treatment and management of various sexual diseases under the caption of
Zoafe bah. In fact, Zoafe bah is a broad term which encompasses different
disease entities like Istirkhae Qazeeb (Erectile dysfunction) Surate Inzaal
(Premature ejaculation) and Qillate Mani (oligospermia) etc. The concept of
Qillate Mani (Qillat- less, Mani-semen) can be correlated with the concept of
oligospermia (Oligo-less, spermia-spermatozoa). The literal meaning of
oligospermia is Qillate haiwane manwiya. The important causes of Qillate mani
are kasrate istifragh, excessive use of mudirrat, sue mizaj of alaate mani,
excessive use of drugs like afyoon (opium) and bhang and excessive riding etc
(Ahmad, 1954; Kabiruddin, YNM; Majusi, 2010; Razi, 2007).
The exact aetiopathogenesis of Qillate haiwane manwiya is not described in
classical Unani literature but on the basis of its cause it can be concluded that
Sue mizaj of alaate mani alter the production of mani due to excessive Baroodat,
Yaboosat, Hararat or Ratubat in alaate mani. For the spermatogenesis, there
is a need of balance in temperament of particular organs. In case of Sue mizaj
of the alate mani the process of spermatogenesis slow down production of
Mani and thus causing oligospermia. (Kabiruddin, YNM)
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17Hippocratic Journal of Unani Medicine
Keeping all these facts in mind, a protocol of an observational clinical trial has
been planned to conduct a preliminary clinical study on Quilatte Haiwane
Manwiya (oligospermia). A commonly used Unani pharmacopoeial compound
drug, Sufoofe Muallif, was selected for the study. Its ingredients of are
Talmakhana, Salab misri, Singhara, Gonde kekar, Mazu sabz, Mastagi Rumi,
Nishasta gandum and Shakar safed (Lateef, 1986). These drugs possess
properties like Muwallide Mani, Mughallize Mani, Muqawwie bah, Mumsik and
Musammine badan which form a rational basis for proposed hypothesis that
the ingredients of selected compound formulation owing to their above cited
properties may be beneficial in patients of oligospermia.
Methodology
The present observational, open uncontrolled clinical study was conducted at
National Institute of Unani Medicine hospital, Bangalore, India. Before initiation
of the project, a comprehensive protocol was framed and put forth for ethical
clearance from the Institutional Ethical Committee. This study was conducted
from February, 2012 to March, 2013. The protocol of the trial was framed for
the study and accordingly findings are recorded on CRF. The inclusion criteria
consisted of being aged from 21 to 50 years infertile males, having sperm
count less than 30 million/ml (Mushtaq, 2007) and willing to follow up and for
semen examination. Those individuals who were suffering with chronic renal
diseases, chronic cardiac ailments, chronic liver diseases, thyroid dysfunctions,
any organic disease of testes and related organs of spermatogenesis, etc or
receiving any medication were not included in the study. Patients fulfilling the
inclusion criteria were given the information sheet having details regarding the
nature of the study, the drug to be used, method of treatment etc. Patients
were given enough time to go through the contents of informed consent sheet.
They were given the opportunity to ask any question and if they agreed to
participate in the study, they were asked to sign the informed consent form.
Eligible patients were selected from OPD of NIUM Hospital, Bangalore. Complete
history and examination including general physical and systemic examinations
were carried out with special attention to endocrine and genital examination,
and recorded on a prescribed proforma which was designed in accordance
with the objectives of the study. Several investigations were carried out with the
aim to exclude the patients with pathological conditions such as Semen Analysis,
Hormone Analysis (Serum Testosterone, FSH, LH), Hb%, TLC, DLC, ESR,
Blood Sugar-F/PP, KFT (Blood urea, serum creatinine) , LFT (SGOT, SGPT,
Alkaline Phosphatase), Urine (Routine and Microscopic). The sample size of
the study was limited to 30 patients and the treatment period was determined
as 60 days. Sufoof Muallif was given to patients in a dose of 5 gram/day for
60 days. Follow up of the patient was done after every 15 days period upto
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18Hippocratic Journal of Unani Medicine
60 days. At each visit patient was inquired about any side effect of the drug
and safety parameters were evaluated before and after treatment. Assessment
of the efficacy of test formulation was carried out on the basis of objective
parameters. Semen analysis (Semen volume, sperm count, sperm motility and
morphology) and hormone analysis (Serum Testosterone, LH, FSH) of each
patient was performed before and after treatment. The objective parameters
were analysed by using Wilcoxon matched-paired signed ranks test, one tail
paired-T test and Kruskal Wallis test.
Trial Formulation (Sufoofe Muallif)
Talmakhana (Asteracatha longifolia) 100 gms
Salab misri (Orchis latifolia) 80 gms
Singhara (Trapa bispinosa) 150 gms
Gonde kekar (Acacia arabica) 150 gms
Mazu sabz (Quercus infectoria) 75 gms
Mastagi rumi (Pistacia lentiscus) 75 gms
Nishasta gundum (Starch) 100 gms
Shakar safed (Sugar) 730 gms
Method of preparation and mode of administration of test drug
Proper identification of the ingredients of the Unani formulation was done by
chief pharmacist, National Institute of Unani Medicine, Bangalore, to ensure
their originality and authenticity. The single drugs then were cleaned by weeding
out unwanted material and impurities and pounded to fine powder.
Results and Observations
Overall 30 patients completed the trial according to the study protocol. Sufoof
Muallif was generally well tolerated and no remarkable adverse events were
reported in the test group. The mean score for sperm count pretreatment (0
day) was 10.53±1.30 while the same post-treatment (60th day) was 18.06 ±
2.57 (p<0.01). The mean and SEM scores for sperm motility on 0 day were
21.70±3.80 and for 60th day were 46.56±5.06 (p<0.01). The mean and SEM
scores for sperm morphology on 0 day were 76.2 ± 6.40 and for 60th day were
91.9 ± 4.22 (p<0.01). The mean and SEM scores for serum testosterone on
0 day were 4.66 ± 0.32 and for 60th day were 4.67 ± 0.32 (p>0.05). The mean
and SEM scores for serum LH on 0 day were 5.8 ± 0.44 and for 60th day were
6.3 ± 0.47 (p>0.05). The mean and SEM scores for serum FSH on 0 day were
6.7 ± 0.89 and for 60th day were 7.08 ± 0.89 (p>0.05).The baseline and after
treatment values of objective parameters are depicted in Table 1.
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19Hippocratic Journal of Unani Medicine
Table 1
Mean ± SEM
Sperm Count Sperm Sperm Serum Serum Serum
(million/ml) Motility Morphology Testosterone LH FSH
BT AT BT AT BT AT BT AT BT AT BT AT
Test 10.53 18.06 21.70 46.56 76.2 91.9 4.66 4.67 5.8 6.3 6.7 7.08
Group ± ± ± ± ± ± ± ± ± ± ± ±
1.30 2.57 3.80 5.06 6.40 4.22 0.32 0.32 0.44 0.47 0.89 0.89
P-Value p<0.001 p<0.001 p<0.01 p>0.05 p>0.05 p>0.05
Figure 1
Discussion
The improvement in the sperm count would be because of Muqqawie bah,
Muwallide mani, Mughallize mani and Musammine badan activities of most of
the ingredients of Test drug i.e. Talmakhana (Asteracatha longifolia), Salab
misri (Orchis latifolia), Singhara (Trapa bispinosa) and Nishasta (Starch). These
results are in conformity with the properties of the drugs as indicated by Unani
scholars such as Ibn Rush’d, Ibn Sina, Hakeem Abdul Hakeem, Najmul Ghani
and Kabeeruddin etc. (Ibn Rushd, 1987; Ghani,YNM; Kabeeruddin, 2010;
Haleem, 2009; Kabeeruddin, 2007). Modern scientific clinical and experimental
studies have also proved aphrodisiac and spermatogenic properties of some
ingredients of Test drug like Asteracantha longifolia, Trapa bispinosa and Orchis
latifolia (Chauhan et al., 2009; Agarwal et al., 2003; Mayank et al., 2008).
Chemical analysis of various test drugs show that apart from different chemical
constituents they possess various nutritional elements such as carbohydrate,
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20Hippocratic Journal of Unani Medicine
protein, minerals like Ca, Mg, Fe, Zn and Cu etc, and vitamins such as thiamine,
riboflavin, pantothenic acid and pyridoxine etc (Patra et al., 2009; Singh et al.,
2010). These chemical constituents and vitamins are essential for the process
of spermatogenesis (Zakai et al., 2011). Therefore, the effect of Test formulation
might be due to presence of these elements. The studies of Tikkiwal et al. and
Wong et al., 2002 reveal that Zinc and folic acids are responsible for
improvement in sperm count and motility.
The improvement in the sperm motility might be due to Muqqawie bah, Muwallide
mani, Mughallize mani properties of the various constituents of test formulation
which have been documented in Unani literature (Ibn Rushd, 1987; Ghani,YNM;
Kabeeruddin, 2007, 2010; Haleem, 2009). Different researches have
documented the antioxidant (Trommer et al., 2005; Kaur et al., 2008; Chryssavgi
et al., 2008) and immunomodulatory (Patel, 2010) activities of Mazu, Mastagi,
Singhara and Samaghe Arabi which are ingredients of Sufoofe Muallif. By
virtue of these antioxidant and immunomodulatory properties test drug is
effective in improving sperm motility. As it has been proved by the studies of
Carmely et al. (2009) and Bansal et al. (2009) etc. that antioxidant and
immunomodulating agents play a major role in improving the process of
spermatogenesis, increasing sperm motility and viablility.
Before and after treatment values of sperm morphology were subjected to
statistical analysis by using Wilcoxon matched-paired signed ranks test and it
was found that the difference between the Mean ± SEM scores of sperm
motility pre and post treatment was statistically significant (p<0.01).
Hormone analysis of each patient was performed for serum testosterone, LH
and FSH. The values of LH and FSH were analyzed statistically by using one
tail paired-T test and it was found that the difference between the Mean ± SEM
scores of serum testosterone was not significant (p> 0.05).
Zakai et al. (2011) mentioned that for the management of oligospermia, the
rational approach is to focus on enhancing those factors which promote sperm
formation which is closely linked to nutritional status. Therefore, it is critical that
men with low sperm counts have optimal nutritional intake. In addition to
consuming a healthful balanced diet, there are several nutritional factors that
deserve special place viz vitamin C and other antioxidants, fats and oils, zinc,
folate, vitamin B, arginine, and carnitine spermatogenesis. As various drugs of
our Test formulation specially Talmakhana and Singhara have Musammine
badan property, and also the chemical analysis proved the presence of various
important nutritional factors like carbohydrate, proteins, minerals and vitamins
such as thiamine, riboflavin, pantothenic acid and pyridoxine etc (Patra et al.,
2009; Singh et al., 2010). The chemical constituents present in the ingredients
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21Hippocratic Journal of Unani Medicine
of Test formulation may further facilitate the process of spermatogenesis and
thereby effective in improving sperm count and motility.
In Unani system of medicine the principle of treatment is based on the concept
of organ protection, strengthening and maintenance of the Quwa (faculties) at
its equilibrium (etedal). The faculties at their equilibrium are balanced inherently
to maintain the normal function of that organ or system. It has been mentioned
that each organ has been gifted with special Quwat for its optimal functioning.
Unsiyaen (Testes) are the azae raesa (vital organs) for Quwwate tanasuliya
(Reproductive power). In case of derangement of function (zoaf) of any organ,
the drugs enhancing its power (muqawwi advia) are advocated. This is the
reason why in Unani system of medicine, for every organ and system there is
a group of tonic drugs (muqawwi advia) proposed that safe guard its larger
interest and bring it near to its equilibrium, if some derangement in its structure
or function takes place. Therefore, most of the sexual diseases are being
treated on the basis of concept of Taqwiyate aaza. The ingredients of Test
formulation are bestowed with the properties like Muqawwie bah, Muwalllide
mani and Muqawwie aam (general tonic) etc, by virtue of these actions these
drugs potentiated the functions of testes. Thus it may be presumed that the
observed significant differences in the values of sperm count and motility
would be due to Muqqawie bah, Muwallide mani and Mughallize mani activities
of the ingredients of Sufoofe Muallif. Thus we can say that scientific studies
and reported effects of individual ingredients of Test drug are in confirmatory
to a great extent with that of our hypothesis as well as the inferences we drew
out of the present study.
In order to determine toxicity of test drug, safety parameter i.e. complete
haemogram, LFT and KFT were carried out before and after the treatment in
each patient. It was found that all the safety parameters were within the normal
levels after the completion of trial. This suggests that Test formulation can be
used safely at mentioned therapeutic dose.
This discussion is helpful to draw the conclusion that the Test formulation is
safe and effective for the management of oligospermia and can be used for
long period without any adverse effect. However, the long term studies with a
larger sample size are required to elucidate other pharmacological actions of
Test formulation.
Study Limitations and strengths:
The main limitation of this trial was the duration of the trial and by longer follow
up; we might reach to more favourable changes in objective parameters. The
strength of this trial is its novelty in the infertile group of males.
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22Hippocratic Journal of Unani Medicine
Conclusion
On the basis of above results and observations it may be concluded that the
Test drug is safe and effective and can be used potentially in the management
of oligospermia. Further long term and large scaled phase III and IV trials are
advocated to explore other important chemicals and pharmacological actions
of the Test formulation.
Acknowledgment
The authors are thankful to Director, National Institute of Unani Medicine,
Bangalore, for providing research facilities to carry out this work.
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23Hippocratic Journal of Unani Medicine
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24Hippocratic Journal of Unani Medicine
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25Hippocratic Journal of Unani Medicine
AAbstract
randomized single blind standard controlled study was
conducted to evaluate the efficacy of Habbe Aftimoon in the patients of
dyslipidemia with Atherosclerosis. Thirty diagnosed patients were selected and
randomly allocated to Control and Test groups (comprising 20 patients in Test
group and 10 in Control group). Habbe Aftimoon in a dose of 4 Habb twice a
day in Test group whereas Lipotab 2 tablets once a day was given in Control
group for 60 days. All the patients were advised low fatty diet and moderate
exercise. Before and after the treatment, both groups were assessed on
subjective and objective parameters. The outcome of treatment were analyzed
statistically by using Paired ‘t’ test, Wilcoxon test, Friedman test with post test,
one way ANOVA with post test and Kruskal Wallis test with Dunn’s multiple
compare test.
The Test drugs exhibited statistically significant result in subjective parameters
(Palpitation and Body weight) in intra group and inter group comparison. In
objective parameters, reduction in S. Cholesterol and increase in L ank ASI
and ABI were observed, and this difference was found statistically significant
in intra group comparison. The control drug exhibited significant improvement
in palpitation and reduces body weight in intra and inter group comparison. In
objective parameters, S. Cholesterol and S. Triglyceride were significantly
decreased in intra group comparison, where as the changes in other subjective
and objective Parameters were remain insignificant in both groups.
The study revealed that Test drug is effective in some objective and subjective
parameters in patients of dyslipidemia with Atherosclerosis particularly in reducing
body weight and S. Cholesterol level. No adverse effect and toxicity was seen
during and after the study. Thus, it can be concluded that Test drug is effective
and safe in the management of Atherosclerosis in dyslipidemic patients up to
some extent. So, it may be recommended for delaying complications of
Atherosclerosis.
Keywords: Atherosclerosis, Dyslipidemia, Habbe Aftimoon, Unani Medicine
Introduction
Tasallube Sharaeen (Atherosclerosis) is one of the commonest conditions
which underlying pathologic process causes several cardiovascular and
cerebrovascular complications. It is well known fact that Hyperlipidaemia and
Obesity are two important risk factors associated with Atherosclerosis (Longo
et al., 2012).
Effect of HabbeAftimoon in thePatients ofDyslipidemiawith TasallubeSharaeen(Atherosclerosis)
1Mohd. Aslam,2*Mohd. Anwar,
3Hamid Ali
and2M. Shoaib
1Department of Moalejat,
Allama Iqbal Unani Medical
College & Hospital,
Muzaffarnagar-251002 (U.P.)
2Department of Ilaj bit Tadbeer,
A.K. Tibbiya College,
Aligarh Muslim University,
Aligarh-202002
3Department of Ilaj bit Tadbeer,
National Institute of Unani Medicine,
Kottigepalaya, Magadi Main Road,
Bangalore-560091
July - September 2014, Vol. 9 No. 3, Pages 25-37
2*Author for correspondence
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26Hippocratic Journal of Unani Medicine
Since very beginning the concept of Tasallube Nabz also exist in Unani system
of Medicine, as most of the ancient Unani scholars like Majoosi, Ibn Sina, Ibn
Rushd, Ibn Zuhr and Samar Qandi have elucidated the cause of narrowing and
stiffness of vessels in their treaties (Ibn Sina, 1993; Khawaja Rizwan, 2010;
Majoosi, 1889; Ibne Zuhr, 1986) Now Tasallube Sharaeen is used as standard
term for arteriosclerosis and atherosclerosis is main type of arteriosclerosis
(Anonymous, 2012).
Atherosclerosis is predicted to become the leading cause of death in India by
2020 (Satishchandra et al., 2011) Tobacco smoking, obesity, hypertension,
diabetes mellitus, elevated plasma homocysteine and LDL are the principal
factors responsible for deposition of lipid in large and medium sized arteries
(Longe, 2002). Slow and progressive lipid deposition narrows down the arterial
lumen by forming atherosclerotic plaque, which initially causes ischemia of the
irrigated organs, but in advance stages results in peripheral vascular diseases,
myocardial and cerebral infarction and stroke etc. (Nicholas et al., 2006).
Now a day many pharmacological and non-pharmacological modes of treatment
are available for prevention of atherosclerosis. Among non-pharmacological,
life style modifications like decreasing daily calorie intake, and increase in
physical activity is indeed helpful for most of the patients but in several
circumstances pharmacological management of atherosclerosis is inevitable.
For this purpose several cholesterol lowering agents such as Levostatin,
Atorvastatin, Simvastatin, Clofibrate, Bezafibrate and Niacin etc are widely
prescribed in conventional Medicine. But long term use of these drugs produces
several adverse effects, such as Hepatotoxicity, Myopathy, Dyspepsia, Renal
failure and Cholelithiasis. (Kumar, 2005; Goodman & Gillman, 2011)
High prevalence of the disease, multi factorial causes and life threatening
complications and most important inability of contemporary system of Medicine
to deliver safe and effective drug for the management of atherosclerosis
effectively, warrant search of alternative treatment to alleviate such a complex
disease of serious complications.
Unani system of Medicine offers different approach of treatment i.e. Ilaj bil
Ghiza (Diet therapy), Ilaj bit Tadbeer (Regimental therapy) and Ilaj bid Dawa
(Pharmacotherapy) (Kabeeruddin, 1954) which are the mainstay of treatment
of hyperlipidaemia and atherosclerosis. These three could be used alone or
in combination. Fundamentally, combination of Ilaj bil Ghiza, and Ilaj bit Tadbeer
are very useful for the prevention of atherosclerosis. The principle of treatment
should be to reduce caloric intake, to burn extra calories deposited in body,
to eliminate Mawade fasida and correction of Sue mizaj barid, use of Qalilul
taghaziya kasirul kammiyat Ghiza along with Riyazate kasira and Hammam
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27Hippocratic Journal of Unani Medicine
(Razi, 1997). Moreover, Unani physician also recommends Ilaj bid Dawa for the
management of Samne Mufrit. A large number of, drugs available in Unani
Medicine which possess action like Muhazzil, Munzij, Mushil, Mufatteh, Jaali
and Muhallil properties could also be used for prevention of Tasallube Sharaeen.
Unfortunately, there is no convincing treatment available for the management
of atherosclerosis in contemporary system of Medicine. Therefore, search of
safe and effective drug for its management is quite necessary. In Unani system
of medicine, Habbe Aftimoon is recommended for the treatment of amraze
balghamia and saudawia. (Akbar Arzani, 2009) The ingredients of Habbe
Aftimoon are Aftimioon (Cuscuta reflexa L.), Gule Surkh (Rosa damascene L.),
Mastagi (Pistacia lentiscus L.), Post Halela Zard (Terminalia chebula L.), Bisfaij
(Polipodium vulgare L.), Ustokhudoos (Levendula stoechas L.) and Namak
Hindi (sodium chloride) (Kabeeruddeen, 2006).
Tasallube Sharaeen is also categorized as amraze saudawia. Thus this drug
may be proven useful in this condition also, but its efficacy has not been
evaluated on scientific parameters particularly for the management of
Atherosclerosis. Therefore, a single blind randomized standard control clinical
trial was designed to evaluate the efficacy of Habbe Aftimoon in the management
of Atherosclerosis in dyslipidemic patients.
Methodology
A single blind standard controlled clinical trial was conducted from March 2012
to January 2013 in National Institute of Unani Medicine Hospital, Bangalore.
The study protocol was designed according to the need of the trial, and
approval was obtained by the Institutional Biomedical Ethics Committee of
NIUM, Bangalore. After providing detailed oral information about the study,
written consent was obtained from the participants. The patients belonging age
of 20–65 years, having dyslipidemia with Tasallube Sharaeen (Atherosclerosis)
were selected for the study. Diagnosis of Atherosclerosis was confirmed by
computerised device (Atherowin & Canwin) on the basis of assessment of right
brachial arterial stiffness index, left brachial arterial stiffness index, right ankle
arterial stiffness index, left ankle arterial stiffness index, right brachial pulse
wave velocity, left brachial pulse wave velocity, carotid femoral pulse wave
velocity and ankle brachial index.
Individuals below 20 years and above 65 years of age , and those having
history of AIDS, Tuberculosis, Hypothyroidism, uncontrolled Diabetes Mellitus,
established I.H.D., advanced Kidney, Liver and Heart diseases and Pregnant
and lactating women were not included to the study. According to subjective
and objective criterion a total of 50 patients were registered for the study from
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28Hippocratic Journal of Unani Medicine
the OPD and IPD of NIUM Hospital. During screening 11 patients did not fulfil
inclusion criteria and excluded from the study, remaining 39 patients were
randomly allocated into Test (Group A) and standard Control (Group B) groups
respectively by using simple randomization sampling method. In the Test group
4 tablets of Habbe Aftimoon twice a day (Each tablet contains 750 mg of Test
drug) was given orally for 60 days whereas Lipotab 2 tab was administered
once a day for the same duration. All the patients were advised low fatty diet
with low caloric value (1600-200 kcal per day) and aerobic exercise for 30-45
minute per day (Agarwal, 2014). All patients were asked to come fortnightly for
the assessment of progression or regression of symptoms. During the
whole duration of protocol concomitant treatment was not allowed in both
groups.
The assessment of efficacy of Test and Control drugs were carried out on the
basis of subjective and objective parameters. Subjective parameters include
symptoms like Palpitation, Xanthelesma, and Nabz sulb which were assessed
at fortnightly, while other objective parameters i.e. body weight, lipid profile,
arterial stiffness, pulse wave velocity and ankle brachial index were measured
before starting treatment and after the completion of treatment.
In order to asses safety of the Test and Control drug , complete Haemogram
(TLC, DLC, Hb%, ESR), Liver Function Test (S. Bilirubin, SGOT, SGPT, Alkaline
Phosphates) and Kidney Function Test (Blood Urea & S. Creatinine) were also
carried out before and after treatment .
During study seven patients from Test group and two patients from Control
group were lost to follow-up, leaving behind 20 patients in Test and 10 patients
in Control group. Therefore, statistical data were calculated on 30 patients
only who were completed entire course of treatment. Data was statistically
analyzed by paired‘t’ test, Wilcoxon matched pair test and Friedman test for
intra group comparison and one way ANOVA and Kruskal-Wallis test with Dunn’s
multiple pair comparison for inter group comparison
Results
Out of 30, 14 (46.66 %) are male and 16 (53.33 %) are female. 05 (25 %),
10 (50%), and 05 (25%) subjects in test group belongs to 20-35 year, 36-50
year, and 51-65 year age groups respectively, similarly 01 (10%), 04 (40%),
and 05 (50%) subjects in standard control group belongs to 20-35 year, 36-
50 year, and 51-65 year age groups respectively (Table 1).
The effect of Test and control drug on various subjective and parameters are
depicted in Table 2 and 3A, 3B.
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29Hippocratic Journal of Unani Medicine
Table 1: Demographic Data
Factor No. of patients Total Percentage
No. of (%)
Test group Control group Patients
Age 20-35 5 1 6 20
36-50 10 4 14 46.7
51-65 5 5 10 33.3
Sex Male 9 5 14 46.67
Female 11 5 16 53.33
Diet Mixed diet 19 9 28 93.33
Vegetarian 1 1 2 6.67
Socio- Upper (I) 0 0 0 0
economic Upper Middle (II) 7 3 10 33.33
status Lower Middle (III) 4 2 6 20
Upper Lower (IV) 8 5 13 43.34
Lower (V) 1 0 1 3.33
Table 2: Effect of Drugs on Subjective Parameter
Parameter Group Assessment day P value
0 day 15th 30th 45th 60th
day day day day
Palpitation Control 3(1,4) 3(1,4) 2.5(1,3) 2(1,3) 1*,#,(1,2)
Test 3(2,4) 3(1,4) 3(1,3) 1a,b,c 1a,b,c
(1,3) (1,3)
Xanthelesma Control 0(0,2) 0(0,2) 0(0,2) 0(0,2) 0(0,2)
Test 0(0,2) 0(0,2) 0(0,2) 0(0,2) 0(0,2)
Nabz sulb Control 3(1,3) 2.5(1,3) 2.5(1,3) 2.5(1,3) 2.5(1,3)
n=10
Test 2(1,3) 2(1,3) 2(1,3) 2(1,3) 2(1,3)
(a) P<0.01 with respect
to zero days test group.
(b) P<0.01 with respect
to 15 day test group.
(c) P<0.01 with respect
to 30 days test group.
* P< 0.01 with respect
to zero day control
group.
# P<0.01 with respect to
15 days control.
After two months administration of Habbe Afteemoon and Lipotab, significant
difference was observed only in body weight, total cholesterol, L ank ASI, and
ABI in Test group, while in standard control group significant difference was
observed only in body weight, total cholesterol and serum triglycerides. Whereas,
no significance difference was observed in other objective parameters neither
in test nor in standard control group.
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30Hippocratic Journal of Unani Medicine
Table 3A: Effect of drugs on objective parameter (20 in Test and 10 in Control group)
Parameters Group Assessment day P value
Before After
Treatment Treatment
Body Control(10) 83.8±4.15 81.9 ±4.2* *→ P<0.01 with respect to before
weight Test(20) 79.0±2.4 76.8±2.6+ treatment in control group
+→ P<0.01 with respect to before
treatment in test group.
S. Control 205±8.93 180±9.35a a→ P<0.05 with respect to before
Cholesterol Test 215±8.73 195±5.93b,c treatment in control group.
b→ P<0.01 with respect to before
treatment in test group.
c→ p< 0.05 with respect to after
treatment in control group.
Triglyceride Control 274±25.7 214±28.9d d→ P<0.01 with respect to before
Test 218±11.3 204±12.6 treatment in control group.
Low Density Control 92.7±11.9 89±10.3 P>0.05 Inter group comparison,
Lipoprotein Test 96.1±9.33 103.1±6.37 with respect to before & after
(LDL) treatment in test & control group.
High Density Control 37.9±2.56 46.4±3.82 P>0.05 Inter group comparison,
Lipoprotein Test 51.1±3.5 44.4±2.32 with respect to before & after
(HDL) treatment in test & control group.
Furthermore, safety markers i.e. Haemogram (TLC, DLC, Hb%, ESR), Liver
Function Test (S. Bilirubin, SGOT, SGPT, Alkaline Phosphates) and Kidney
Function Test (Blood Urea & S. Creatinin) remained normal before and after
treatment. (Table 4)
Discussion
Tasallube Sharain (Atherosclerosis) is one of the commonest causes of the
premature vascular diseases, causing Ischemic Heart Disease, Cerebrovascular
accidents, Stroke and Hypertension. Obesity and Hyperlipidaemia are two risks
factor which are associated with atherosclerosis. In modern system of medicine
several Hypolipidaemic agents are being used for the prevention of
atherosclerosis. However, these drugs are neither drug of choice of
Atherosclerosis nor producing convincing therapeutic effects. Furthermore, the
side effects of these Hypolipidaemic agents are also causes of concern.
In view of the above facts, the development of Hypolipidaemic agents from
herbal sources is quite necessary. There is no dearth of such drugs in Unani
System of Medicine. Habbe Aftimoon is one of the compound formulations
which is effective in the treatment of Amraze Saudawia. Apart from this the
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31Hippocratic Journal of Unani Medicine
Table 3B: Effect of drugs on objective parameter (20 in Test and 10 in Control group)
Parameters Group Assessment day P value
Before After
Treatment Treatment
Right Brachial Control 30±4.77 28.9±2.87 P>0.05 Inter group comparison,
Arterial with respect to before & after
Stiffness Index Test 24.4±1.66 23.3±2.31 treatment in test & control group.
(R Bra ASI)
Left Brachial Control 27±3.06 28.14±3.39 P=0.924 Inter group comparison,
Arterial with respect to before & after
Stiffness Index Test 28.04±2.42 28.07±3.77 treatment in test & control group.
(L Bra ASI)
Right ankle Control 42.08±3.29 42.08±2.82 P=0.66 Inter group comparison,
Arterial with respect to before & after
Stiffness Index Test 38.68±3.23 37.53±2.38 treatment in test & control group.
(R Ank ASI)
Left ankle Control 33.9±4.19 39.4±3.3 e’!P<0.05 with respect to before
Arterial treatment in test group. Inter group
Stiffness Index Test 31.5±2.04 35.9e±2.54 comparison, p=0.09
(L Ank ASI)
Right Brachial Control 2344±359 2004±216 P=0.75 Inter group comparison,
Pulse Wave with respect to before & after
Velocity Test 1592±267 1966±180 treatment in test & control group.
(R ba PWV)
Left Brachial Control 1116±367 1426±188 P>0.05 Inter group comparison,
Pulse Wave with respect to before & after
Velocity Test 1058±295 1645±238 treatment in test & control group.
(L ba PWV)
Carotid Control 1209±213 1229±142 P=0.62 Inter group comparison,
Femoral Pulse with respect to before & after
Wave Velocity Test 870±18 1270±119 treatment in test & control group.
(C F PWV)
Ankle Control 1.11±.031 1.10 ±.031 f’!P<0.05 With respect to before
Brachial treatment in test group.
Index (ABI) Test 1.01±.029 1.10f±.021
ingredients of Habbe Aftimoon possess some important pharmacological
properties such as Hypolipidaemic, Antioxidants, Mohazzil actions. These
pharmacological actions are effective in the delaying process of Atherosclerosis.
Therefore, A single blind standard control study was designed to evaluate the
efficacy of Habbe Aftimoon in the patients of Tasallube Sharain (Atherosclerosis).
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32Hippocratic Journal of Unani Medicine
Table 4: Effect of Test and Control drugs on Safety parameters
Parameters Test Control
No=20 No=10
B.T A.T B.T A.T
Hb% gm% 12.41±.31 12.46±.33 12.34±.41 11.92±.44
TLC cells/cu 8185±330 8062±386 8290±438 7170±480
DLC P 57.6±1.6 57.3±1.4 58±1.5 59.4±2.45
L 37.9±1.4 37.6±1.2 35.7±1.48 35.1±2.32
Cells/cu DLC E 3.2±.18 3.5±0.23 4±.29 3.5±0.22
M 2.2±.20 2.3±.21 2.3±0.21 2.3±0.21
Cells/cu B 0±0.00 0±0.00 0±0.00 0±0.00
ESR (mm/1hrs) 27.4±4.2 27.5±4.6 21.5±4.18 23.6±5.20
FBS (mg/dl) 99±2.98 103±3.96 101±7.8 103±7.62
PPBS (mg/dl) 167±12.3 157.7±9.36 151±16.7 146±16.2
B. Urea (mg/dl) 31.3±1.6 29.1±1.5 29.3±2.42 35.5±2.75
S. Creatinin (mg/dl) 0.89±.02 0.84±0.03 0.85±0.02 0.94±0.05
S. Bilirubin (mg/dl) 0.64±0.05 0.65±0.05 0.52±0.04 0.52±0.05
SGPT(IU/L) 29.5±2.5 31.6±3.82 19.4±3.59 20.5±1.68
SGOT (IU/L) 25.1±1.8 23.7±1.6 20.5±1.7 18.7±2.34
Alkaline Phosphates 131±6.5 121±4.9 139±5.5 132±6.9
(IU/L)
After 60 days treatment, significant improvement has been noticed in subjective
and objective parameters such as S. Cholesterol, Left ankle Arterial Stiffness
Index (L Ank ASI), Arterial Brachial Index (ABI), and body weight significantly in
test group and total cholesterol, triglycerides, and body weight in standard
control group respectively in atherosclerotic patients.
From above, it is evident that test drug is effective in intra group comparison.
Such effect may be due to ingredients of test drug Habbe Aftimoon which
contain antioxidant, Anti-atherosclerotic and Antioxidant properties. (Cheng et
al., 2003; Naik et al., 2006; Lee et al.,1639; Naik et al., 2004; Chang et al.,
2010; Selvaraj et al., 2007) and hypolipidaemic properties of Halela zard
(Chang et al., 2010) Mastagi (Stella et al., 2009; Xiuzhen Han et al., 2007;
Dedoussis et al., 2004, Duke 2008) and Ustukhodoos (Nikolaevskii et al.,
1990; Yumi et al., 2008; Parejo et al., 2002; Ferreira et al., 2006; Gulcin et al.,
2004).
This improvement may be due to Muhallil (Resolvent), Mulattif (Demulcent)
and Munzij balghame wa sauda properties of Ustukhudoos (Kulkarni et al.,
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33Hippocratic Journal of Unani Medicine
2004; Ghani, 2010; Nabi, 2007) Mufatteh, Muhallil and Mushile Balgham wa
sauda properties of Aftimoon, Bisfaij, GuleSurkh, Mastagi, and Halela zard.
(Ghani, 2010; Nabi, 2007; Ibn Sina, 1993) These findings are in accordance
with the description given by Razi, Ibne Sina, Ibne Baitar, N.Ghani, Mohd. Azam
khan etc. Further, some recent studies revealed that Mastagi and Ustukhodoos
possess anti atherosclerotic action. (Duke, 2008; Catherine et al., 2001)
Individual drugs that constitute the ingredients of Test drugs have been reported
to possess some interesting pharmacological effects that directly or indirectly
support our contentions regarding the efficacy of the Test drugs. Afteemoon
Bisfaij, Halela zard, Ustokhudoos, Mastagi possesses Mufatteh sudad, Muhallil,
Mulattif and Mushile sauda wa balgham properties. (Ghani, 2010) Ustokhodoos
(Catherine et al., 2001) and Halela zard (Selvaraj et al., 2007; Duke, 2002)
possesses Anti-arteriosclerotic and hypolipidaemic properties, Halela zard
(Prajapati et al., 2005, Cheng et al., 2003) Gule Surkh, (Prajapati et al., 2005;
Said, 1997; Boskabady, et al., 2001) and Mastagi (Benhammou et al., 2008;
Tassou et al., 1995) possesses antioxidant and cardiotonic properties. These
effects are in the same line, as we have mentioned above that the drugs are
producing effects because of hypolipidaemic, antioxidant and cardio tonic
properties. Thus, on the basis of the scientific studies and the reported effects
of the individual ingredients of Test drugs are in conformity to a greater extent
with that of our hypothesis as well as the inferences we drew out of the present
study.
In the light of above discussion, it can be concluded that the Test drugs
produced significant hypocholesterolemic and anti obesity effect without
demonstrating any sign of toxicity or adverse effect.
Although, the Test drug did not produces any significant effect in most of the
objective parameters except body weight, S. Cholesterol, L Ank ASI, and ABI.
However, the Test drug exhibited improvement in some objective parameter
such as on Serum Cholesterol, body weight. Obesity and hypercholesterolemia
are considered an important risk factor for development of atherosclerosis.
The Test drug (Habbe Aftimoon) is quite effective in reducing body weight and
serum Cholesterol level, thereby it may play pivotal role in delaying of
atherosclerosis process. Therefore, it can be concluded that Test drug can be
used for the prevention of atherosclerosis and delaying the progression of the
disease. As a matter of fact, there is no curative treatment available in any
system of medicine. The conventional system of medicine is using hypolipidaemic
and thrombolytic agents for the prevention of atherosclerosis. (Goodman and
Gillman, 2011) Hence, the Test drug Habbe Aftimoon can be safely used for
the same purpose.
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34Hippocratic Journal of Unani Medicine
Conclusion
On the basis of above result and discussion, it can be concluded that the
compound formulation Habbe Aftimoon is effective in reducing lipid profile in
the patients of atherosclerosis associated with dyslipidemia. Hence, this drug
could be effectively used for prevention of atherosclerosis and to reduce
progression of its manifestation. Since the diverse mechanism is involved in
the development of Atherosclerosis and disease is complex in nature. Therefore,
some elaborate studies are required to ascertain other pharmacological action
of Test drug relatively for longer duration.
Acknowledgement
The authors are very much thankful to Director of National Institute of Unani
Medicine, Bangalore, for providing necessary facilities and conducive
environment in the Institute to carry out this research work
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37Hippocratic Journal of Unani Medicine
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38Hippocratic Journal of Unani Medicine
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39Hippocratic Journal of Unani Medicine
HAbstract
yperlipidaemia is an important factor for the
development of atherosclerotic lesion which is responsible for various
complications such as ischemic heart diseases, cerebrovascular diseases and
hypertension etc. These complications are major cause of mortality and
morbidity. The association of Hyperlipidaemia with development of atherosclerotic
lesion has prompted the researchers of various field of medicine to develop
safe and efficacious drug for the management of Hyperlipidaemia. Although, a
number of plant origin hypolipidaemic drugs have been screened for
hypolipidaemic activity but none of them offers convincing treatment. In Unani
system Medicine a large number of drugs are reported to possess possible
treatment for some metabolic diseases particularly obesity (Saman-e-Mufrat)
and its related complications. Most of these drugs have not been subjected for
evaluation on scientific parameters. Keeping these facts in mind, a single blind
standard controlled non randomized trial was planned to evaluate the efficacy
of a Roghane Kalonji (Nigella sativa oil) in the management of primary
hyperlipidaemia.
The present study was conducted on 60 diagnosed patients of primary
hyperlipidaemia at Ajmal Khan Tibbiya College Hospital, Aligarh Muslim University
Aligarh. The patients were allocated in two groups. Group A (Test group)
comprising of 40 patients and Group ‘B’ (Placebo group) comprising of 20
patients. In group ‘A’ the Roghane Kalonji in the dose of 6 ml twice a day was
given whereas in group ’B’ Atrovastatin in the dose of 10 mg once a day was
administered for 3 months. The subjective and the objective parameters (lipid
profile) were assessed on 0, 15th, 30th, 60th, 90th days. The test drug was
found to be statistically significant in lowering serum cholesterol and serum
triglyceride level in the patients of Hyperlipidaemia (p<0.05).
Keywords: Hyperlipidaemia, Roghan-e-Kalonji, Lipid profile.
Introduction
Hyperlipidaemia is a major public health problem worldwide. It accounts about
3.2 million visits to a doctor per year. It is characterized by an excess of fatty
substances such as cholesterol, triglycerides and lipoproteins in the blood. It
is an important risk factor for development of atherosclerosis and ischaemic
heart diseases. More than half of the coronary heart diseases are attributed
to the abnormalities in lipids and lipoproteins metabolism. It also reflects the
adverse impact of the sedentary life style and dietary factors like dietary fat
Efficacy ofRoghan-e-Kalonji (Nigellasativa oil) in theTreatment ofPrimaryHyperlipidaemia
1M. Nazim,1*B.D. Khan,
1Misbahuddin Siddiqui
and2M. Shoaib
1Department of Moalijat2Department of Ilaj-bit-Tadbeer
A.K. Tibbiya College,
Aligarh Muslim University,
Aligarh-202002
July - September 2014, Vol. 9 No. 3, Pages 39-48
1*Author for correspondence
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40Hippocratic Journal of Unani Medicine
intake greater than 40% of the total calories, saturated fat intake greater than
10% of total calories. (Akbar et al., 1930; Biff et al., 2003; Fauci et al.,2008;
Hongdao, 2006; Joshep, 2003; Jurjani, 1878; Khan et al., 2002, Masson et al.,
2003; Rath et al., 1991). In classical Unani text Shaham is broadly classified
into two types Samin and Riwaj. It is essential for the nourishment of the body
and essential for normal health. According to Unani concept, when the amount
of lipid increase in the blood it leads to increased viscosity and stickiness
(ghilzat and Lazojat) of blood that reduces lumen of vessels which results in
Tasallube Sharaeen. (Maseehi, 1963; Ibne Sena, 1929; Razi, 1999).
As such, there is no direct reference to this disease per se, but hyperlipidemia
is usually associated with obesity. The ancient Unani physicians like Buqrat
(460 BC) and Ibn-Sina (980-1037 AD) have described Saman-e-Mufarat in
their Lexicon. They have mentioned the etiological factors, symptoms
complications like paralysis, constriction of vessels, coma and sudden death
owing to obesity in their clinical observations. Due to the lack of diagnostic
facilities and means of evaluation of lipids in blood, ancient Unani physicians
may have considered obesity and hyperlipidaemia as one disease thus
describing obesity as a whole not specifying excess of fat in blood (Al-Qamri
Mansoori, 1255H; Aqsarai, 1907; Arzani, 1954; Baitar, 1999; Hussain, 1980;
Ibnul-Qaf, 1986; Jalinoos, 1903; Majoosi, 1889; Mansoor Ibne Mohammad,
1989; Maseehi, 1963; Razi, 1999; Rushd, 1987; Ibne Sena, 1929).
The association of hyperlipidaemia with development of atherosclerotic lesion
has prompted the researchers of various field of medicine to develop safe and
efficacious drug for the management of Hyperlipidaemia. Although, a number
of plant origin hypolipidaemic drugs have been screened for hypolipidaemic
activity but, none of them offers convincing treatment. Even though, in mainstream
Medicine Statin (HMG-co Reductase inhibitor) is being used but long term
administration is associated with several side effects. (Lazar et al., 2011; Siig
et al., 2004; Ziajka, 1998) Therefore, search of safe and effective drug is
imperative. In Unani system medicine a large number of drugs are reported to
be used for the treatment of obesity (Saman-e- Mufrat) and its related
complications. Such as Zanjbeel, (Rafiquddin, 1985), Muqil (Anonymous, 2001;
Tripathi, 1984), Kundur (Anonymous, 2004) etc., among them Kalonji (Nigella
sativa L.) (Rafiquddin, 1985) is one the important drugs, which is extensively
used for the remedy of many diseases. The Holy Prophet said that black seed
(kalonji) is remedy for all diseases except death. (Ibne-al-Qayyum, 1985) Recent
studies revealed that seed oil contains major active constituent Thymoquinone
which posses anti inflammatory, antioxidant, anti hypolipidaemic and cardio
protective properties. Nigella sativa oil contains omega-3 fatty acids and other
polyunsaturated and monounsaturated fatty acids in large amount (Ali, 2003).
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41Hippocratic Journal of Unani Medicine
Consumption of some Omega-3 fatty acids such as Eicosapentaenoic (EPA)
and Decosahexaenoic (DHA) acids present in fish oil has shown a preventive
action against cardiovascular diseases (Anonymous, 2010).
Some practicing Unani physicians are using Kalonji oil in the treatment of
cardiovascular diseases and dyslipidaemic with better results, but clinical efficacy
of Kalonji oil has not been carried out, so far, particularly in relation to
hypolipidaemic property. Keeping these facts in view, a single blind standard
controlled non randomized clinical trial was planned to evaluate efficacy of a
Roghane Kalonji in the management of primary hyperlipidaemia.
Materials and Methods
For the assessment of efficacy of the Roghan-e-kalonji in the management of
primary hyperlipidaemia, patients were selected from Ajmal Khan Tibbiya College
Hospital OPD, Aligarh, during the period 2010-2012. In the present study, the
patients who attended OPD with the symptoms of palpitation, chest pain, joints
pain, obesity, dyspnoea on exertion, xantholesma and were enrolled for the
screening of hyperlipidaemia. The patients belonging to the age group of 20-
60 years of either sex, ready to participate in the study and whose serum
cholesterol, serum triglyceride level was found abnormal, were included in the
clinical study. The patients of secondary hyperlipidaemia such as hypothyroidism,
diabetes mellitus, nephrotic syndrome and obstructive liver diseases were
excluded from the study on the basis of relevant symptoms and investigations.
Similarly, the patients using oral contraceptive pills and having history of chronic
alcoholism were also excluded from the study. Diagnosis of hyperlipidaemia
was confirmed on the basis of history, clinical examination and analysis of Lipid
profile markers i.e. S. Cholesterol, Triglyceride, VLDL and HDL.
Study Procedure
After complete physical general, systemic examination and biochemical
investigations, patients who fulfilled all the inclusion criteria and signed written
consent, were included in the clinical trial. Total 60 patients were selected for
the study. The patients were allocated into two groups, i.e. Test group (40
patients- Group A) and control group (20 patients- Group B). In the group A
the Test drug was administered in the dose of 6 ml three times a day for a
period of 90 days, while in the group B the Atrovastatin was given in the dose
of 10 mg once a day for same duration. Assessment was done on 0, 15th, 30th,
60th, 90th day of treatment for subjective and objective parameters. In all
patients lipid profile (Serum Cholesterol, Serum Triglyceride, and HDL
cholesterol) was carried out before and after treatment. The data was statistically
analyzed by using Students t test.
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42Hippocratic Journal of Unani Medicine
In order to assess toxicity of the drugs, safety parameters like liver function
Test (Serum Bilirubin, AST, ALT and Alkaline Phosphate), Kidney Function Test
(Blood Urea, Serum Creatinine) and complete Haemogram were also carried
out.
Results
In this study out of sixty patients of hyperlipidaemia 19 patients were 25-35
years of age, 19 patients were 36-45 years of age, 12 patients were 46-55
years of age, 07 patients were 56-65 years of age group and 03 patients were
>65years of age. The highest prevalence was found in 4th decade. The
percentage of female patients is 76.66% was slightly higher than the male
patients i.e. 23.33%. The demographic data and other observation are depicted
in Table 1.
The effects of test drug on objective parameters i.e. Lipid profile (Serum
Cholesterol, Serum Triglyceride, HDL, LDL, and VLDL) and body weight are as
follows.
Table 1: Demographic Data of patients in Test and Control group n=60(40+20)
N Fp% N Fp%
Age group Mizaj
25-35 19 31.66% Balghami 46 76.66%
36-45 19 31.66% Damvi 08 13.33%
46-55 12 20.0% Safravi 06 10.0%
56-65 07 11.66% Saudavi 0 0%
>66 03 5.0%
Gender Marital Status
Male 14 23.33% Married 53 88.88%
Female 46 76.66% Unmarried 07 11.66%
Religion Dietary Habit
Muslim 50 83.33% Vegetarian 09 15.0%
Hindu 10 16.66% Mixed Diet 51 85.0%
Occupation History of
Students 03 5.0% Xanthomata 07 11.66%
Service 09 15.0% Xanthelesma 13 21.66%
Labour 02 3.33% Arcus cornea 07 11.66%
Business 17 28.33%
House wife 29 48.33%
History of IHD Family History of HLD
Present 35 58.33% Present 28 46.66%
Absent 25 41.66% Absent 32 53.33%
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43Hippocratic Journal of Unani Medicine
Effect on Serum Cholesterol
In Test group mean serum cholesterol level was 183.32 mg/dl ±31.40 before
treatment and at the end of study it was167.60 mg/dl ±26.52, showing mean
reduction 15.72 mg/dl ±4.88 and which was found to be significant (P<0.001)
(Table 2).
Effect on Serum Triglyceride
In Test group mean serum triglyceride level was 299.07 mg/dl ±97.64 before
treatment and at the end of study it was 235.82±82.92 mg/dl, showing mean
reduction 63.25 mg/dl ±14.72 and which was found to be significant (P<0.001)
(Table 2).
Effect on HDL
In Test group mean HDL level was 33.67 mg/dl ±6.96 before treatment and at
the end of study it was 38.08 mg/dl ±6.45, showing mean elevation 4.41 mg/
dl ±0.51 and which was found to be significant (P>0.05) (Table 2).
Effect on LDL
In Test group mean LDL level was 89.83 mg/dl ±33.83 before treatment and
at the end of study it was 82.35 mg/dl ±29.17, showing mean reduction 7.48
mg/dl ±4.66 and where was found to be significant (P<0.05) (Table 1).
Effect on VLDL
In Test group mean VLDL level was 59.81 mg/dl ±19.52 before treatment and
at the end of study it was 47.16 mg/dl ±16.58, showing mean reduction 12.65
mg/dl ±2.94 and which was found to be significant (P<0.001) (Table 2).
Table 2: Effect of Test drug on Objective parameter in Test and control group
S. Parameter Group-A n=40 Group-B n=20
No.
Before After P-Value Before After P-Value
Treatment 90 days Treatment 90 days
(Base line) (Base line)
1 S. Cholesterol 183.32±31.40 167.60 ±26.52 <0.001 185.20 ±29.27 141.60±17.53 <0.001
2 S. Triglyceride 299.07±97.64 235.82±82.92 <0.001 290.40 ±58.41 195.25 ±39.12 <0.001
3 HDL 33.67±6.96 38.08±6.45 >0.05 39.42±6.93 40.15±6.34 >0.05
4 LDL 89.83±33.83 82.35 ±29.17 <0.05 87.69±22.87 62.40 ±17.63 <0.001
5 VLDL 59.81±19.52 47.16±16.58 <0.001 58.08±11.68 39.05±7.82 <0.001
6 Body weight 67.27±4.22 64.45±3.73 <0.001 67.00±4.40 65.20±4.67 <0.001
7 SBP 136.22±10.18 131.30±8.10 <0.001 142.60±6.96 133.4±6.55 <0.001
8 DBP 86.75±6.65 84.25±5.21 <0.001 86.30±6.33 83.40±4.55 <0.05
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44Hippocratic Journal of Unani Medicine
Effect on Body weight
In Test group mean WHR level was 67.27±4.22 kg before treatment and at the
end of study it was 64.45±3.73 kg, showing mean reduction 2.82±0.49 kg and
which was found to be significant (P<0.001) (Table 2).
Effect on Safety Parameters
The safety parameters of the test drug like, AST, ALT, Blood Urea, Serum
Creatinine, Hb% and ESR were remained within the normal limits before & after
treatment, in both groups (Table 3).
Discussion
Hyperlipidaemia is a major risk factor for the development of atherosclerotic
heart disease. Reducing plasma cholesterol level particularly LDL cholesterol
reduces the risk of coronary heart disease and other associated complications.
Indeed, dietary modification plays an important role for the prevention of
atherosclerotic diseases but in certain circumstances use of hypolipidaemic
drugs are imperative. Although, a large number of drugs are being used in
contemporary systems of Medicine but search of new plant origin hypolipidaemic
drug is still thrust area of research. The seed of Nigella sativa L. (black seed)
and its oil have been used since long time for the treatment of many diseases
including hyperlipidaemia in traditional system of medicine. The Holy Prophet
Muhammad (PBUH) said, “The black seed (kalonji) is the remedy for every
disease except death” (Ibne-al-Qayyum 751H). Recent experimental studies
revealed that this drug possess lipid lowering effects in dyslipidaemic patients.
Further studies exhibited that Nigella sativa oil (Roghan-e-Kalonji) contains
Table 3: Effect of Test drug on Safety parameter in Test and control group
S. Parameter Group-A n=40 Group-B n=20
No.
Before After Before After
Treatment 90 days Treatment 90 days
(Base line) (Base line)
1 S. Bilirubin 0.98±0.2083 0.94±0.1585 1.053±0.23 1.14±0.17
2 AST 23.13±4.66 23.74±4.23 23.49±3.72 25.35±4.60
3 ALT 21.77±3.87 22.44±3.29 20.56±4.27 23.02±6.59
4 S. Alkaline 120.80±15.84 122.07±12.85 122.0±10.57 131.0±13.88
Phosphates
5 Blood Urea 29.22±7.51 26.31±5.91 26.86±5.45 27.84±4.75
6 S. Creatinine 0.95±0.144 0.90±0.108 0.93±0.13 0.94±0.11
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45Hippocratic Journal of Unani Medicine
chemical constituents such as polyunsaturated fatty acids, Omega-3 Fatty
acids, Eicosapentaenoic acid (EPA), Decosahexaenoic acid (DHA) and Alpha
Linolenic acid (ALA) (Anonymous, 2010).
Several preclinical studies revealed that Omega-3 fatty acids decrease the
triglyceride levels either by decreasing hepatic synthesis or secretion of VLDL
particles by inhibiting various enzyme transcription factors or EPA and DHA
increase the activity of lipoprotein lipase, leading to an increase in chylomicron
clearance. (Harris et al., 2008)
Furthermore, researches on Nigella sativa seed oil (Roghan-e-kalonji) revealed
that the seed oil contains major active constituent Thymoquinone. Much of the
biological activity of the seed has been possess due to Thymoquinone, which
shows antioxidant, anti-inflammatory, anti-hyperlipidaemia and cardioprotective
properties. (Ali, 2003; Badary et al., 2000).
The present study demonstrates that the Test drug (Nigella sativa oil) significantly
reduces. Serum Cholesterol, Serum Triglyceride, LDL Cholesterol. These findings
of the study are in accordance to the previous experimental studies as reported
by the Dakha Khani et al., 2000 who revealed that the administration of Nigella
sativa oil for four weeks duration showed significant decrease in serum
Cholesterol, Triglycerides and increases of HDL Cholesterol. Further in other
clinical study administration of 2.5ml of Nigella sativa oil in morning and evening
produce significant hypolipidaemic effects (Dakha Khani et al., 2000).
This drug also exhibited significant effect in reducing Blood pressure and other
subjective parameters. The safety parameters like, AST, ALT, Blood Urea, Serum
Creatinine, Hb% and ESR were remained within the normal limits before & after
treatment, in both groups. This indicates that oral administration of the test
formulation is safe for therapeutic use.
The above mentioned effect of Test drug are mainly due to chemical constituent
present in the seeds of Kalonji particularly Thymoquinone and omega -3 Fatty
acid and unsaturated Fatty acids. Further antioxidant, anti-inflammatory,
hypolipidaemic and cardioprotective properties facilitate the effect of Test drug
in the patients of hyperlipidaemia (Ali, 2003).
Conclusion
In the present study the Test drug was found to be significant in lowering
serum cholesterol and serum triglyceride level in the patients of hyperlipidaemia
without producing any adverse effect. Therefore, it can be concluded that the
Test drug possesses significant hypolipidaemic effect and thus it can be used
as a safe and cost effective therapy in the management of hyperlipidaemia.
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46Hippocratic Journal of Unani Medicine
However, further study is required to explore hidden potential of Test drug on
larger population.
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49Hippocratic Journal of Unani Medicine
HAbstract
yperlipidemia is a major public health problem
throughout the world. Consequent atherogenic disorders occupied the first
place in five major killer diseases due to high mortality and high morbidity in
the world.
Adouble blind, placebo controlled, randomized clinical trial was conducted to
test the safety and efficacy of a unani formulation in subjects with hyperlipidaemia.
Unani formulation significantly improved lipid profile as compared to placebo.
During 60 days therapy, no noticable adverse/side effectes were detected in
both treatment groups.
Keywords: Hyperlipidaemia; Badranjboy; Bombyx mori; Abresham;
Cardioprotective
Introduction
Hyperlipidemia is considered as lifestyle disorder in present age and it is a
major public health problem throughout the world. It is characterized by
increased lipids in blood due to either increase in rate of synthesis or decrease
in rate of breakdown of lipoproteins (Clayton et al., 1999). Hyperlipidemia is a
common condition which may either results from primary abnormality in lipid
metabolism or is a secondary manifestation of some other conditions (Christofer
et al., 2004). Worldwide, the prevalence of hyperlipidaemia is about 39,000
per 100,000 patients. In developed countries, the prevalence of hyperlipidaemia
is about 57,000 per 100,000 patients. In developing countries, the prevalence
of hyperlipidaemia is about 26,000 per 100,000 patients (Michael Gibson,
2013). Epidemiological evidence suggests 1% increase in Coronary Heart
Disease (CHD) risk for each 1% increase in Low Density Lipoprotein (LDL), 2-
3% reduction in CHD risk for each 1% increase in High Density Lipoprotein
(HDL) (Annonymous, 2001). According to the available data, the atherogenic
disorders occupied the first place in the five major killer diseases due to high
mortality and high morbidity in the world. The diseases are usually considered
as disease of atherogenic pathology and affect the various parts of the body
viz. ischemic heart disease, cerebrovascular accidents and peripheral vascular
disorders etc. Various epidemiological studies suggest that the development of
atherosclerosis and ischemic heart disease is strongly associated with
hyperlipidemia.
To overcome the problem of hyperlipidaemia, day by day several synthetic
drugs of better efficacy are being introduced in the modern system of medicine.
ClinicalEvaluation of aUnaniFormulation forthe Treatmentof Fart-e-Tadassum-Fid-Dam (Hyprer-lipidaemia): Arandomized,Double Blind,PlaceboControlledClinical Study
1Rais-ur-Rahman,2Afshan Qaiser
and3*Yasmeen Shamsi
1Department of AYUSH,
Ministry of Health and F.W.,
Government of India,
GPO Complex, INA,
New Delhi-110023
2Department of Moalijat,
A&U Tibbia College, Karol Bagh,
New Delhi-110005
3Department of Mahiyatul Amraz,
Faculty of Medicine (U),
Jamia Hamdard,
New Delh-110062
July - September 2014, Vol. 9 No. 3, Pages 49-55
3*Author for correspondence
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50Hippocratic Journal of Unani Medicine
In this series nicotinic acid was the first drug to be introduced by the Altschul
et al in 1955 (Rudolf et al., 1960). Use of hypolipidaemic agents and low fatty
diet is the corner stone of the management of hypercholesetrolaemia. Several
hypolipidaemic drugs have been already introduced in main stream medicine
such as Hydroxy methyl glutaryl CO-A (HMG CO-A) reductase inhibitors
(levostatin, atorvastatin), bile acid sequestrents (colestipol) and fibric acid
derivetives (gemfibrozil, and fenofibrates) etc. But the long term use of these
drugs causes various side effects like myalgia, arthragia, dyspepsia and
cholelithiasis loss of libido, impotence etc (Tripathi, 1994). Such side effects
limit the use and the efficacy of these drugs. Hence, there is a need to develop
a drug from herbal source which should be safe, cost effective, easily available
and efficacious. Keeping in view all the above mentioned drawbacks of modern
medicine in the management of hyperlipidaemia, need was felt to conduct a
clinical trial with dried aqueous extract of Abresham (Bombyx mori), Badranjboya
(Nepeta hindostana) and Arjun Bark (Terminalia arjuna) in the ratio of 1:2:1.
Preclinical study of this unani formulation has already been conducted on
isoproterenol treated rats, which showed remarkable hypolipidemic and
cardioprotective effects of test drug comparable to control (Tajuddin et al.,
2006 & 2007).
Materials and Methods
This study was double blind, randomized, placebo controlled clincal trial ,
carried out in the department of Moalejat , A &U Tibbia College, Karol Bagh,
New Delhi, from September 2012 to March 2013. The aim of the study was to
evaluate the efficacy and safety of unani formulation in the treatment of Fart-
e-Tadassum Fiddum (Hyperlipidaemia) on scientific parameters.
Study Drug
The study drug was a combination of three Unani drugs supplied by Dehlvi
Naturals, India, in the form of capsule. Placebo capsules were also supplied by
Dehlvi Naturals, India, the composition of test drug is given in Table-1.
Table 1: Composition of Unani Formulation
Each 500 mg capsule contains dried aqueous extract of:
Unani Name Scientific Name Part Used Quantity
Abresham Bombyx mori Raw Silk Cocoon 125 mg
Badranjboya Nepeta hindostana Whole Plant 250 mg
Arjun Terminalia arjuna Bark 125 mg
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51Hippocratic Journal of Unani Medicine
Partcipants
Eligible subjects as per the inclusion/exclusion criteria were enrolled in the
study after obtaining written informed consent according to Helsinki declaration.
Inclusion Criteria
Subjects (men and women) aged 18-65 years were eligible for the study if they
had a history of dyslipidemia regardless of strict diet control and had fasting
LDL-Cholesterol= 130-159 mg/dl, Total Cholesterol= 200-239 mg/dl,
Triglycerides= 150-190 mg/dl and HDL-Cholesterol: <40 mg/dl (Anonymous,
2001).
Exclusion Criteria
The exclusion Criteria were pregnancy, lactation, intake of oral contraceptives
or any other medication that might affect serum lipids (thyroid or steroid
hormones, beta blockers, diuretics etc), H/O cardiovascular disease, impaired
hepatic, renal function, malignancy, secondary hyperlipidaemia and body mass
index >30 kg/m2.
Treatment
At baseline, lipid profile determinations and laboratory safety tests were
performed and the eligible cases as per the inclusion/exclusion criteria were
randomly assigned to receive either drug or placebo capsule in the dose of 1
capsule twice daily for a period of 60 days. All the patients were instructed to
maintain low cholesterol diet. Clinical examination and laboratory tests were
done at each visit. Adverse events were recorded and compliance with study
medications was assessed at each follow up visit.
Primary Outcome Measures
The primary efficacy end point was percentage reduction from baseline in
LDL-Cholesterol, Triglycerides and Total-Cholesterol.
Secondary Outcome Measures
Percentage of change from baseline in HDL, LDL/HDL ratio and TC/HDL ratio.
were the secondary outcome measures.
Safety Assaessment
For the assesment of safety, Liver function test, Kidney function test, Haemogram
and ECG were carried out at baseline, on first follow up visit i.e., on on 15th
day of treatment and at the end of therapy i.e., on 60th day. Data from the
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52Hippocratic Journal of Unani Medicine
physical/clinical examination, laboratory tests and interview for adverse events
as recorded in CRF were included in the analyses of safety and tolerability.
Statistical Analysis
The changes between pre-treatment and post treatment values of primary and
secondary outcome obtained in test group were compared with those obtained
in placebo group by using unpaired ‘t’ test. Statistical calculations were performed
with GraphPad InStat statistical softwere version 3.10 . Statistical analysis was
done only for those patients who completed the course of treatment for 60
days.
Results
Total 70 patients were registered out of them 10 cases (4 receiving test drug
and 6 receiving placebo) were dropped out from the study, only 30 patients in
test group and 30 in the placebo group completed the treatment..
Pretreatment and post treatment (after 60 days) maean values of lipid profile
components are their percent changes areshown in Table-2 & Figure-1.
Sixty days treatment with test drug was significantly effective than placebo on
the primary efficacy measures in reducing LDL-C by 15.98% as compared with
4.21% in the placebo group (p < 0.001). Test drug also significantly reduced
total cholesterol (TC) and triglycerides by 10.30% (compared with 0.94% in
placebo group) and 12.05% (compared with 1.77% in placebo group)
respectively (p value was <.0001 in both cases). The mean reduction in total
lipids was 10.24%, in drug group, compared with placebo group (0.64%).
The test drug was also found significantly effective than placebo on the
secondary efficacy measures in reducing LDL/HDL ratio by 26.87% compared
with 2.47% in the placebo group and TC/HDL ratio by 24.22% (compared with
6.27% in placebo group). A significant rise in HDL was observed in test group
(10.57%) compared to placebo group (3.00%), (p<0.001) respectively (p value
was <.0001 in both cases).
Safety and Tolerability
Unani formulation (also placebo) treatment for 60 days did not impair physical
safety indicators such as body weight, pulse rate or blood pressure.
Laboratory safety indicators e.g., kidney function test (blood urea, serum
creatinine), Liver function test (ALT, AST, serum bilirubin, serum alkaline
phosphalase) and haemogram remained within the normal limits in all study
patients.
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53Hippocratic Journal of Unani Medicine
Figure 1: Mean Percent Change in Various Components of Lipid Profile After Treatment
Table 1: Effects of Test Drug (Unani Formulation) and Control (Placebo) on Lipid Profile
Lipid Profile Mean±SEM Mean±SEM % t p
0 Day 60th Day Change value value
LDLCholesterol (mg/dl)
Test Drug 145.23±1.61 122.02±2.2 15.98% 2.69 0.001
Control 140.30± 2.3 134.38±4.04 4.21%
Total Cholesterol (mg/dl)
Test Drug 217.80±1.63 195.37±1.77 10.30% 7.28 0.0001
Control 216.97±1.82 214.93±1.64 0.94%
Triglycerides (mg/dl)
Test Drug 171.01±1.50 150.40±1.63 12.05% 5.138 0.0001
Control 163.97±1.75 161.07±1.84 1.77%
Total Lipids
Test Drug 607.3±4.07 545.13±3.82 10.24% 7.77 0.0001
Control 598.23±3.77 594.40±5.06 0.64%
HDLCholesterol (mg/dl)
Test Drug 38.23±0.84 42.27±0.87 10.57% 2.206 0.001
Control 40.87±1.54 42.10±1.39 3.00%
LDL/HDL ratio (mg/dl)
Test Drug 3.87±0.14 2.83±0.10 26.87% 2.056 0.04
Control 3.24±0.13 3.16±0.13 2.47%
TC/HDL ratio
Test Drug 5.78±1.94 4.38±2.03 24.22% 3.832 0.02
Control 5.1±1.18 4.78±1.17 6.27%
N=30 in each group; LDL=Low Density Lipoprotiens; HDL= High Density Lipoprotiens;
TC=Total Cholesterol, SEM=Standard error of Mean
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54Hippocratic Journal of Unani Medicine
Discussion
In the present clinical trial the effects of test drug have been tested on all the
components of lipid profile in a double blind, randomized fashion and the
safety of the drug has also been established. Unani formulation (test drug)
significantly reduced LDL,TC, triglycerides, LDL/HDL and TC/HDL comparable
with control(placebo). Unani formulation also significantly improved HDL level
than that observed in control group. Individual ingredients of unani formulation
have been reported to possess important pharmacological actions that directly
or indirectly support our contention regarding efficacy of test drug on human
being. Abresham, Arjun chhaal and Badranjboya possess diverse
pharmacological activities like cardio tonic, anti inflammatory, anti-oxidant,
anxiolytic, fibrinolytic, and anti-platelet activities as evident by previous
pharmacological studies (Collabawalla, 1951; Ghani, ynm; Mahdi et al., 2011;
Singh et al., 2001; Maulik et al., 1997).
The effect of unani test formulation on various components of lipid profile
could be due to hypolipidaemic, cardiotonic and cardiac stimulant activities of
Arjun (Terminalia arjuna) and cardiotonic, anti hypercholesterolemic and
antiatherogenic effects of Abresham (Bombyx mori) & Badranjboya (Nepeta
hindostan) (Monahan, 2007; Halleys Khan et al., 2011; Ghani, ynm).
All these support the cardiovascular protective effects of test drug. Both
Abresham (Bombyx mori) and Arjuna (Terminalia arjuna) have been reported
to have potent antioxidant activity; these findings also support cardioprotective
effects of test drug. Badranjboya (Nepeta hindostana) is known to prevent
myocardial infarction which supports its lipid lowering action.
The results of present study suggest that the unani test formulation is safe and
efficacious in treating hyperlipidaemia.This formulation can be valuable in
prevention of atherosclerosis and cardiovascular disease by anti-platelet,
fibrinolytic, anti-oxidant and cholesterol lowering activities of its ingredients.
Conclusion
The results of this study can be concluded as:
• The unani formulation significantly reduced LDL Cholesterol, Total
Cholesterol, Triglycerides, LDL/HDL ratio, and TC/HDL ratio as compared
with placebo.
• Improvement in HDL Cholesterol obtained with unani fornulation
treatment was significantly greater than that of placebo.
• The unani test formulation was well tolerated and no adverse/side
effect were observed during the entire period of protocol therapy.
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55Hippocratic Journal of Unani Medicine
References
Anonymous, 2001. Executive summary of the Third Report of the National
Cholesterol Education Program (NCEP) Expert panel on detection, evaluation
and treatment of high blood cholesterol in adults (Adult treatment panel III),
JAMA may 16; 285 (19): 2486 -97.
Christofer R, Edwin A, Nicholas R. Nikkia, 2004. Davidson’s principles and
practice of medicine, 19th edition. Churchill living stone, pp.308-311.
Clayton, L., Thomas, Taber’s Cyclopedic, 1999. Medical Dictionary, 16th Ed.
F.A. Davis Company, USA, pp.863-1039.
Collabawalla, H., 1951. An evaluation of the cardiotonic and other properties
of Terminalia arjuna. Indian Heart Journal 3:205-230.
Ghani Najmul, ynm. Khzainul Advia: Sheikh Mohd. Basher and Sons Pub. Urdu
Bazar Lahore, pp.209, 217.
Halleys Khan, Z.M., Hossain Md. Faruquee , Md. Munan Shaik, 2011.
Phytochemistry and Pharmacological Potential of Terminalia arjuna L.
Medicinal Plant Research, 3(10:70-77.
Maulik, G., Maulik, N., Bhandari, V., 1997. Evaluation of antioxidant effectiveness
of a few herbal plants. Free Radical Research 27: 221-228.
Michael Gibson, C., 2013. Hyperlipidemia epidemiology and demographics.
Wikidoc Editor-In-Chief: C. Michael Gibson, Hardik Patel ib.wikidoc.org/
Hyperlipidemia epidemiology and demographics.
Mir Mahdi Ali, Arumugam, A., Sarasa, Bharati, 2011. Effect of crude extract of
Bombyx mori cocoons in Hyperlipidemia and atherosclerosis. Journal of
Ayurveda and Integrative Medicine 2(2):72-78.
Monahan, 2007. Effect of Hyperlipidemia on autonomic and cardiovascular
control in human. Journal of Applied Physiology 103(1): 162-169.
Rudolf Altschul, Abram Hoffer, 1960. The effect of nicotinic acid on
hypercholesterolaemia. Canad. M.A.J. 82:783-785.
Singh, K.P. and Jayasomu, R.S., 2001. Bombyx mori; A review of its potential
as medicinal insect. Pharmaceutical Biology 39:1-5.
Tajuddin, Nasiruddin, M. and Ahmad, N., 2007. Cardioprotective effect of Unani
formulation in rats. Indian Journal of Traditional Knowledge 6(4):663-667.
Tajuuddin, Nasiruddin M., Ahmad, N., 2006. Effect of a unani formulation on
lipid profile in rat. Indian J. Pharmacol 38(1):56-57.
Tripathi, K.D., 1994. Essentials of Medical Pharmacology, 3rd edition. Jaypee
Brothers Medical Publishers, pp.59345.
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56Hippocratic Journal of Unani Medicine
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57Hippocratic Journal of Unani Medicine
IAbstract
n view of probability of adulteration especially in unorganized
drugs, this study was designed for standardization of Dammul Akhawain
(Dragon’s blood) to generate data for future reference. The study was carried
out on a samples of Dragon’s blood obtained from the plant Pterocarpus
marsupium Roxb, considered as the standard sample. The study comprised of
morphology, physicochemical study, physical constants, preliminary
phytochemical study and spectrophotometery. Since in literature parameters
for this have not been mentioned sufficiently, therefore, the findings of this
study may be considered as standard for quality assessment of available
sample of Dragon’s blood.
Keywords: Pharmacognosy, Unorganized drug, Standardization, Phytochemistry.
Introduction
Adulteration, substitution, misidentification, quality inconsistency, and controversy
pertaining to herbal drugs are challenging the wide acceptability of traditional
systems of medicine. Therefore, it is imperative to determine authenticity of
crude drugs used for the preparation of medicaments, because it is associated
with the safety of consumers. However, it becomes challenging when two or
more plants are claimed to be the source of one drug especially in case of
unorganized drugs, such as gum, resin, oleo-resin and oleo-gum-resin, as in
such circumstances herbarium and drug museum, which are the main sources
of information, become of little use (Bonakdar, 2002). Eventually, important
drugs compromise their efficacy in spite of possessing significant effects.
Among the various steps to be taken for solving these problems, quality
assessment of samples of all herbal drugs appears to be of utmost importance.
Conventional methods of standardization substantiated with analytical techniques
are considered most reliable tools for quality assessment of herbal drugs
(Shinde, and Dhalwal, 2007). Most of the regulatory guidelines also suggest
macroscopic, microscopic, physicochemical and phytochemical standardization
of medicinal plants materials. HPLC, HPTLC, and spectrophotometery etc. may
further reinforce the above methods (Xing et al., 2010).
Dragon’s blood (DB), as known in trade, is a bright red resin obtained from a
number of different taxa such as Croton, Dracaena, Daemonorops, Calamus
and Pterocarpus (Shinde, Dhalwal, 2007). Hence, a number of plants such as
Croton draconoides Müll Arg, Pterocarpus marsupium Roxb, Calamus rotang
Linn, Croton draco Schltdl & Cham, Croton lechleri Müll Arg, Croton urucurana
PharmacognosticEvaluation ofDammulAkhawain withReference toStandardization
Ehteshamuddin,
*Abdul Wadud,
Ghulamuddin Sofi,
Mohammad Yusuf Ansari
and
Shamim Irshad
National Institute of Unani Medicine,
Kottigepalya, Magadi Main Road,
Bangalore-560091
July - September 2014, Vol. 9 No. 3, Pages 57-66
*Author for correspondence
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58Hippocratic Journal of Unani Medicine
Baill, Croton xalapensis Kunth, Daemonorops draco Blume, Daemonorops
didymophylla Becc, Daemonorops micranthus Becc, Daemonorops motleyi Becc,
Daemonorops rubra (Reinw ex Blume) Mart, Daemonorops propinquus Becc,
Dracaena cinnabari Balf.f, Dracaena cochinensis Hort ex Baker, and Pterocarpus
officinalis Jacq etc. have been accounted as the source of DB (Xing et al.,
2010). Such a big list of sources for one drug creates enormous degree of
uncertainty. Even if two or more sources, as happens in case of some drugs,
are considered the problem persists as it is not clear which source the available
sample belongs to.
DB is a red resin and the name refers to reddish resinous product applied to
many red resins described in literature (www.en.wikipedia.org, 2010). However,
red gum resin of some of the above mentioned sources have the official status
in their respective countries which they are indigenous to, such as red gum
resin of Pterocarpus marsupium Roxb., family Fabaceae is known as Indian
Dragon’s blood (www.thefullwiki.org, 2010), red gum resin of Dracaena cinnabari
is considered as Socotra Dragon’s blood. Similarly, Dracaena cochinensis is
said to be the source of Chinese Dragon’s blood (www.aseanbiodiversity.info,
2010).
DB is an important drug used in Unani Medicine as Qabiz (astringent), Habis
(styptic), Muqawie meda (stomachic) , Mohallil-e-Auram (resolvent), Dafe-Zaheer
(anti dysentery) (Ibn Sina, 2007). Its liniment is useful in anal fissure, prolapsed
rectum and inflammation of rectum. It is styptic and astringent for stomach
whether used alone or with other astringent drugs, when used as enema it
causes constipation (Khan,1314 AH).
In this study, red gum- resin of Pterocarpus marsupium Roxb. (DB) has been
considered as the standard sample viewing that the same is commonly available
in Indian crude drug markets. Therefore, it was obtained from the plant
Pterocarpus marsupium Roxb, and was evaluated pharmacognosticaly to
establish standard parameters for further studies.
Materials and Methods
Materials
The sample of DB was collected from Pterocarpus marsupium Roxb. grown in
Erepalya, Bidadi, Hobli, District Ramnagar, Karnataka, India. It was authenticated
by Dr. Shiddamallayya N and Dr. V. Rama Rao, vide Drug Authentication/
SMPU/NADRI/BNG/2010-11/961. A voucher specimen has been deposited in
the Drug Museum of National Institute of Unani Medicine (NIUM), Bangalore,
vide Voucher specimen No.01/IA/Res./2011.
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59Hippocratic Journal of Unani Medicine
Methods
Organoleptic evaluation
The organoleptic characters like color, odor, taste, luster, texture, fracture,
consistency and cut surface of were examined by naked eye (Anonymous,
1968).
Physicochemical evaluation
For estimation of extractive values and ash values, methods described in
British Pharmacopoeia (Anonymous, 1968) were applied. Moisture content was
determined by TGA method (Anonymous, 1992).
Determination of pH value
The pH value of 1% and 10% aqueous solution was estimated by the method
described by (Khandelwal, 2008; Brewster and Mcewen, 1971).
Determination of melting point
Melting point was estimated by melting point apparatus model C-LMP-1,
Campbell electronics.
Solubility test
Solubility was tested by the method described in British Pharmacopoeia
(Anonymous, 1968).
Qualitative Phytochemical evaluation
For preliminary phytochemical studies, powder of the DB was extracted in
different solvents viz. petroleum ether, di-ethyl ether, chloroform, ethanol,
acetone, benzene and distilled water. The extracts were subjected to various
qualitative phytochemical tests for estimation of alkaloids, glycosides,
carbohydrates, phenol compounds, tannins, phytosterols, proteins and amino
acids etc.
Alkaloid was tested by Dragendroff’s test, Mayer’s test, Hager’s test and
Wagner’s test (Anonymous, 1992). Protein and amino acids were tested by
Ninhydrin test, Biurette’s reaction, Million’s reaction and Xanthoproteinic reaction
(Khandelwal, 2008; Brewster, and Mcewen, 1971). Glycosides were tested by
Molish’s test (Paech and Tracey, 1955). Cardiac glycosides were tested by
Keller-killiani test. Bufadenoloids were tested by Liebermann’s test. Flavonoids
were tested by Ammonia test (Anonymous, 1992). Saponin was tested by
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60Hippocratic Journal of Unani Medicine
Honey comb frothing test (Arthur and Chan, 1962). Tannins were tested by
Ferric chloride test (Brewster and Mcewen, 1971). Phenols were tested by
Ferric chloride test and Lead acetate test (Khandelwal, 2008). Phytosterols /
Terpenes were tested by Hosse’s reaction, Liebermann Burchard’s reaction,
and Moleschott’s reaction (Khandelwal, 2008).
Test for Inorganic constituents
Ash of DB was prepared. To the ash 50% v/v hydrochloric acid and 50% v/v
Nitric acid were added, and kept for an hour and then filtered. Various tests
were performed with the filtrate for qualitative estimation of inorganic constituents
such as sulphate, phosphate, iron, chloride, carbonate and nitrate (Brewster
and Mcewen, 1971).
Spectrophotometery
The alcoholic extract was subjected to Spectrophotometery by using UV-VIS
Spectrophotometer. The test was performed at room temperature with the
following settings: Number: 18-1885-01-0259; Spectral Bandwidth: 2.00 nm;
scan Range: 190.00 to 360.00 nm; Measure Mode: Abs; Interval: 1.00 nm,
Speed: Medium.
Results
The results of macroscopic evaluation are shown in table 1 and figure 1&2.
The extractive values taken in ethanol, chloroform, diethyl ether, pet. ether,
benzene, acetone, and distilled water; mean percentage of total ash, acid
insoluble ash, water insoluble ash and water soluble ash; moisture content as
obtained by TGA method; mean percentages of pH value in 1% and in 10%
aqueous solution, and the melting point are given in table 2. The Preliminary
Table 1: Morphology of Dragon’s blood
S.No. Characteristics Result
1. Color Yellowish red
2. Luster Lustrous
3. Fracture Transverse
4. Texture Brittle
5. Consistency Liquid
6. Odor Odorless
7. Taste Astringent
8. Cut surface Smooth
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61Hippocratic Journal of Unani Medicine
Phytochemical screening of the different solvent extracts was done systemically
for phytochemical constituents. Glycoside, amino acid and phytosterol were
positive. Iron, nitrate and phosphate were also detected. Spectrum scanning
gave three peaks and three valleys (Figure 3).
Discussion
Due to crude nature of herbal drugs, traders often take advantage of it and
indulge in malpractices; however, it may happen due to ignorance as well.
Usually it is noticed that commercial samples of some drugs do not match with
their description in literature (Kartik et al., 2010).
Figure 1: Samples of Dragon’s blood Figure 2: Powder of dried sample of
Dragon’s blood
Table 2: Physical constants of Dragon’s blood
S.No. Parameters Values (Mean ±SEM)
1. Extractive Solvents Pet. ether 0.00
Values Di-ethyl ether 0.00
Chloroform 0.00
Benzene 0.00
Acetone 28.26 ± 0.86
Ethanol 63.33± 1.46
Distilled water 68.44±2.25
2. Ash Values Total Ash 4.03±0.19
Acid insoluble Ash 2.59±0.14
Water insoluble Ash 12.94±0.16
Water soluble Ash 0.45±0.08
3. Moisture content 0.00%
4. pH 1% solution 4.66± 0.17
10% solution 4.78 ± 0.05
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62Hippocratic Journal of Unani Medicine
Table 3: Solubility of Dragon’s blood in different organic solvents at different temperature
S.No. Solvents Result
Temperature°C
20 40 60 80
1. Ethanol Soluble
2. Pet. Ether Insoluble
3. Di-ethyl ether Insoluble
4. Chloroform Insoluble
5. Benzene Insoluble
6. Acetone Insoluble
7. Distilled water Insoluble
So literature survey can give preliminary ideas about probable sources of
drugs to draw some conclusion. Most of the literature consulted revealed that
gum-resin of Pterocarpus marsupium (www.en.wikipedia.org), Daemonorops
draco (www.botanical.com), Dracaena cinnabari (Al-Awthan et al., 2010), and
Dracaena cochinensis (www.aseanbiodiversity.info, 2010; www.aseanbiodiversity.
info, 2010) may be considered as the sources of DB for different countries. It
Figure 3: Spectrum Scan curves of Dragon’s blood
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63Hippocratic Journal of Unani Medicine
was also concluded that gum-resin of Pterocarpus marsupium Roxb is the
Indian Dragon’s blood.
Morphological studies of crude drugs give some idea at very first sight. Color,
odor, taste, luster, fracture, and consistency etc. (Evans, 2008; Pearson and
Prendergast, 2007) are some prominent characters, therefore, these characters
were observed.
It is important to note that physical constants of a drug are good criteria for
identification. These constants are extractive values, ash values, and moisture
content. These parameters are widely accepted for checking purity of drugs
(Anonymous, 1992). These parameters too were applied in this study.
The constituent of a particular drug sample can be estimated in terms of %
with respect to air dried weight by extracting in various solvents known as
extractive value which was applied to the sample. In literature gum resin of
Pterocarpus marsupium has been reported to be 80 - 90 % soluble in alcohol.
Our finding demonstrated no yield in diethyl ether, pet. ether and benzene.
However the yield % was 28.26 ± 0.86, 63.33± 1.46, 68.44±2.25 in ethanol,
acetone and distilled water, respectively.
Ash value is taken in terms of total ash, acid insoluble, water insoluble and
water soluble ash. In some cases there may be considerable difference in total
ash value in the same drug which may either be due to variation in the amount
of oxalate or some adulteration with metallic and the like materials or earthy
material. In such cases acid in soluble ash is taken into consideration. This
parameter was also applied in our study.
The water content (moisture) of crude drugs is another important parameter
for checking purity of drugs. In this study, Thermo gravimetric Analysis (TGA)
method was applied for estimation of moisture content. This method is suitable
for all types of substances as it provides quantitative measurement of mass
change in materials. It records changes in mass from dehydration, decomposition
and oxidation of a sample with time and temperature (El-Sayd and Makawy,
2010). We found 0.00% moisture in the sample.
DB is a gum-resin and for these types of drugs, melting point, pH and solubility
may be considered important parameters. Gum-resin is insoluble in water and
petroleum ether but more or less soluble in alcohol, chloroform, and ether.
Crude drugs containing mixed chemicals are described with certain range of
melting points. pH of solution of a substance at 1 % w/v and 10% w/v of water
soluble portion can give accurate estimation of purity of a drug. These
parameters were also. No data regarding melting point and pH of gum resin
of Pterocarpus marsupium are available in literature; therefore, we considered
our findings as stand. In literature melting point of gum resin of Calamus draco
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64Hippocratic Journal of Unani Medicine
is shown to be 76°C. (www.henriettesherbal.com, 2010). The melting point of
standard sample did not coincide with the reported finding. Similarly, pH and
solubility were estimated at 20, 40, 60, and 80°C. Except solubility, no data on
gum- resin of Pterocarpus marsupium, which is 80-90 % soluble in cold water
and almost soluble in alcohol, are available (Kokate, 2007), we considered our
results as standard.
The analysis of physiologically active compounds is important parameter for
checking the authenticity of a drug. These compounds are alkaloids, glycosides,
flavonoid, phytosterol, essential oil, resin, tannin, etc. These were also estimated.
Analysis of inorganic constituents may also be considered parameter for
checking the authenticity of drug. In literature Pterocarpus marsupium is shown
to contain flavonoids, tannins, and phytosterol. Our findings are in confirmation
with the report.
Spectrophotometery may be a sophisticated tool for standardization of crude
drugs. Spectrum scanning curves were obtained to get preliminary information.
It demonstrated peaks and valleys of different absorbance and wave length,
indicating presence of different constituents.
Conclusion
Detailed data on DB regarding physical, chemical and other properties are not
available to compare our findings, therefore, our findings may be considered
as standard for Indian Dragon’s blood for future reference.
Acknowledgement
The authors are thankful to the Director, National Institute of Unani Medicine,
Bangalore, for providing facilities for experimentation.
References
Al-Awthan,Y.S., Abu Zarga, M. and Shtaywy A., 2010. Flavonoids Content of
Dracaena cinnabari Resin and Effects of the Aqueous Extract on Isolated
Smooth Muscle Preparations, Perfused Heart, Blood Pressure and Diuresis
in the Rat. Jordan J Pharmaceuti Sci. 3 (1):8 -17.
Anonymous, 1968. British Pharmacopoeia, General Medical Council.
Pharmaceutical Press, London, pp.l1276-77; 1285- 88.
Anonymous, 1992. Quality Control Methods for Medicinal Plant Materials, rev.
1, Organisation Mondiale De La Sante. World Health Organisation, p.159.
Arthur, H.R. and Chan, R.K.R., 1962. A Survey of Hong Kong Plants testing for
Alkaloids, Essential oil, and Saponin. Trop-sci. 4: 147.
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65Hippocratic Journal of Unani Medicine
Bonakdar, R.A., 2002. Herbal cancer cures on the Web: noncompliance with
the Dietary Supplement Health and Education Act. Family Medicine 2002,
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El-Sayd, N.I., and Makawy, M.M., 2010. Comparison of Methods for Determination
of Moisture in Food, Research J Agriculture and Biol Sci. 6(6): 906-11.
Evans, W.C., 2008. Trease and Evans Pharmacognosy, Elsevier, A Division of
Reed Elsevier India Private Ltd., New Delhi, pp. 57-60, 525.
Ibn Sina, 2007. Al-Qanun Fit Tib. Part 1st, Vol.2nd. (Urdu translation by Kantoori,
H.G.). Idara Kitab-ul-Shifa, New Delhi, p. 80.
Kartik, C.P., Surendra, K.P., Ranjit, K.H., Kumar, K.J., 2010. Traditional
Approaches towards Standardization of herbal Medicines-A review. J
Pharmaceut Sci and Technol 2 (11): 372-379.
Khan, M.A., 1314H. Muheete Azam, Part 2, Vol. 2nd. Matabae Nizami, Kanpur,
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Springer & Verlag Gottingen. Hidelbrg, Berlin,pp. 295,334,471.
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66Hippocratic Journal of Unani Medicine
www.thefullwiki.org/Dragon%27s_blood#wikipedia_Name_and_source. [cited
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67Hippocratic Journal of Unani Medicine
LAbstract
iver diseases have become one of the major causes of
morbidity and mortality in man and animals all over the globe and hepatotoxicity
due to drugs appears to be the most common contributing factor. About 20,000
deaths occur every year due to liver disorders. Hepatocellular carcinoma is
one of the ten most common tumors in the world with over 2, 50,000 new cases
each year. Plants have potent biochemical components of phytomedicine. Plant
based natural phyto-constituents can be derived from any morphological part
of the plant and may contain active components. The beneficial medicinal
effects of plant materials typically result from the combinations of secondary
products present in the plant. The medicinal actions of plants are unique to a
particular plant species or groups and are consistent with this concept as the
combination of secondary products in a particular plant is taxonomically distinct
as such different types of drugs such as acetaminophen; chloroquinine and
isoniazid are inducing hepatoxicity in the world. Herbal plants have been used
traditionally by herbalists worldwide for the prevention and treatment of liver
disease so. Herbal drugs were prescribed even when their biologically active
compounds were unknown because of their effectiveness, few side effects and
relatively low cost.
In this review, an attempt has been made to compile the reported
hepatoprotective activity of plants from India and abroad and may be useful to
develop evidence based medicine to cure different kind of liver diseases in
man and animal.
Keywords: Hepatic disorder, Hepatoprotective herbs, Ayurvedic formulations
Introduction
Liver has a pivotal role in regulation of physiological processes. It is involved
in several vital functions such as metabolism, secretion and storage.
Furthermore, detoxification of a variety of drugs and xenobiotics occurs in liver.
The bile secreted by the liver has, among other things, an important role in
digestion. Liver diseases are among the most serious ailment. They may be
classified as acute or chronic hepatitis (inflammatory liver diseases), hepatosis
(non inflammatory diseases) and cirrhosis (degenerative disorder resulting in
fibrosis of the liver). Liver diseases are mainly caused by toxic chemicals
(certain antibiotics, chemotherapeutics, peroxides oil, aflatoxin, carbon-
tetrachloride, chlorinated hydrocarbons, etc.), excess consumption of alcohol,
infections and autoimmune/disorder.
HepatoprotectiveActivity ofExtracts andChemicallyDefinedMolecules fromHerbal Drugs :Review#
1*Manoj Kumar Pandey,2Nitin Rai
and2Rajeev Kr. Sharma
1Pharmacopoeia Commission for
Indian Medicine, Raj Nagar,
Ghaziabad-201002
2Pharmacopoeial Laboratory for
Indian Medicine, Kamla Nehru Nagar,
Ghaziabad-201002
July - September 2014, Vol. 9 No. 3, Pages 67-90
#Invited Paper1*Author for correspondence
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68Hippocratic Journal of Unani Medicine
The liver functions as a great metabolic factory and is particularly concerned
with metabolizing drugs, especially those given orally. It plays a key role in the
metabolism of lipids, proteins and carbohydrates, as well as in
immunomodulation. The sheer complexity and varied nature of its interactions
continually expose it to a variety of toxins, therapeutic agents etc., making it
susceptible to literally hundreds of diseases. Some of these diseases are rare;
others are common, such as hepatitis, cirrhosis, pediatric liver disorders, alcohol
related disorders, liver cancer, and weakened liver function on older people.
Cirrhosis is the third leading cause of death in adults aged between 25 and
59, and seventh leading cause of death overall. It has been estimated that
approximately 14 - 16 million people are infected with this virus in South East
Asia region and about 6% of the total population in the region are carriers of
this virus.
The situation in India is more serious so far as viral hepatitis is concerned. It
is reported that one among every 25 Indians is a carrier of hepatitis B virus
and fifth major cause of mortality of people in the age groups of 15-45 years.
1% of total adult death attributed in India is due to infection caused by hepatitis
virus B. Besides, incidence 60% chronic liver diseases and 80% of primary
liver cancer are due to residual effects of hepatitis B infection. With of lack of
safe and effective treatment for liver diseases, researches have been turned
towards alternative therapies with ethnic drugs of herbal origin used traditionally,
especially in lights of new findings.
Treatment of liver diseases is still largely influenced by holistic approach in
different system of medicines. The modern allopathic has very limited effective
remedies. However much of remedies claims to be available in folk lore
traditional system of medicine based on plants. More than 600 numbers of
plants based commercial products are available in different parts of world
market for the treatment of variety of liver diseases. In India alone there are
more than 60 poly herbal preparations available in market.
In vivo and in vitro investigations established conclusively that many such
plants species does posses prophylactic and therapeutic activity. Recent
development of both in vivo and in vitro investigation procedure laid the
foundation for scientific exploration of such plants species as well as helped
in validating the folk lore claims. This is evident from the voluminous scientific
publications on such investigations on traditional herbal remedies particularly
in last two decades. Isolation of novel active phytoconstituents from many such
plant species possessing significant potency of antihepatotoxic, will lead for
further development of ideal remedies for various liver diseases.
Hence, there is an ever-increasing need for safe hepatoprotective agents.
Herbal-based therapeutics for liver disorders have been in use in India for a
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69Hippocratic Journal of Unani Medicine
long time and popularized world over by leading pharmaceuticals. Despite the
significant popularity of several herbal medicines in general, and for liver
diseases in particular, they are still unacceptable as treatment modalities for
liver diseases. The limiting factors that contribute to this eventuality are lack
of standardization of the herbal drugs, identification of active ingredient(s)/
principles(s), randomized controlled clinical trials (RCTs) and toxicological
evaluation.
A large number of plants and formulations have been claimed to have
hepatoprotective activity. Nearly 160 phytoconstituents from 101 plants have
been claimed to possess liver protecting activity. In India, more than 87 plants
are used in 33 patented and proprietary multi ingredient plant formulations.
About 600 commercial herbal formulations with claimed hepatoprotective activity
are being sold all over the world. However, only a small proportion of
hepatoprotective plants as well as formulations used in traditional medicine are
pharmacologically evaluated for their safety and efficacy
Numerous plants and polyherbal formulations are used for the treatment of
liver diseases. However, in most of the severe cases, the treatments are not
satisfactory. Although experimental evaluations were carried out on a good
number of these plants and formulations, the studies were mostly incomplete
and insufficient. The therapeutic values were tested against a few chemicals-
induced subclinical levels of liver damages in rodents. Even common dietary
antioxidants can provide such protection from liver damage caused by oxidative
mechanisms of toxic chemicals. However, experiments have clearly shown that
plants such as Picrorrhiza kurroa, Andrographis paniculata, Eclipta alba, Silibum
marianum, Phyllanthus maderaspatensis and Trichopus zeylanicus are
sufficiently active against, at least, certain hepatotoxins.
Single plant may not have all the desired activities. A combination of different
herbal extracts/fractions is likely to provide desired activities to cure severe
liver diseases. Development of such medicines with standards of safety and
efficacy can revitalise treatment of liver disorders and hepatoprotective activity.
The traditional medicinal plants species have been subjected in various
experimental models of investigation and attempt has made to calibrate their
therapeutical activity. The herbs used in hepatic disease have been extensively
exploited all over the world and large numbers of the plant species has been
documented as hepatoprotective antihepatotoxic, cholegoge and choleric. A
diverse nature of chemical compounds has been identified from such plants
species through bio-assay guided investigation. However, in some plant species
the total extract or fraction of extract has reported to possess better and
potent biological activity compared to isolated pure compound(s) from the
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70Hippocratic Journal of Unani Medicine
extracts. Plant derived extracts and chemically defined molecules are
enumerated in Table 1 and 2.
Table 1: Plant Extracts with Hepatoprotective Activity
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
Acanthaceae India Leaves Alcohol In vivo CCl4 Babu et al.
Acanthus ilicifolius L. (2001)
Andrographis lineata India Leaves Aqueous, In vivo CCl4 Sangameswaran
Nees methanol et al. (2008)
Andrographis India Leaves Alcohol In vivo CCl4 Rana and
paniculata (Burm.f.) Avadhoot (1991)
Nees
Anisotes trisulcus Yemen Whole Ethanol In vivo CCl4 Fleurentin et al.
(Forssk.) plant (1986)
Asteracantha Sri Whole Aqueous In vivo CCl4 and Hewawasam
longifolia L. Lanka plant PCM et al. (2003)
Hygrophila auriculata India Seeds Methanol In vivo PCM and Singh and Handa
(K.Schum.) Heine Thioacetamide (1995)
Hypoestes triflora Rwanda Leaves Aqueous In vivo CCl4 Van Puyvelde
(Forssk.) Roem. and et al. (1989)
Schult
Rhinacanthus nasuta India Root Methanol In vivo CCl4 Suja et al. (2003)
(L.) Kurz.
Adoxaceae Viburnum Southern Leaves Aqueous- In vivo CCl4 Mohammed et al.
tinus L. Europe methanol (2005)
Aizoaceae Trianthema India Leaves Ethanol In vivo PCM and Kumar et al.
portulacastrum L. Thioacetamide (2004)
Apiaceae Apium India Seeds Methanol In vivo PCM and Singh and Handa
graveolens L. Thioacetamide (1995)
Carum copticum L. Pakistan Seeds Aqueous- In vivo CCl4 and PCM Gilani et al.
methanol (2005a)
Apocynaceae China, Leaf Aqueous In vivo CCl4 and GAIN Xiong et al.
Apocynum venetum L. Japan (2000)
Araliaceae Taiwan Whole Methanol In vivo CCl4 and Lin and Huang
Acanthopanax plant Acetami- (2002)
senticosus (Rupr. and nophen
Maxim.) Harms
Asclepiadaceae India Stem Ethyl In vivo CCl4 Sethuraman et al.
Sarcostemma bark acetate (2003)
brevistigma Wight
Asteraceae Argentina Aerial Aqueous In vivo Bromobenzene Kadarian et al.
Achyrocline parts (2002)
satureioides (Lam.)
DC.
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71Hippocratic Journal of Unani Medicine
Artemisia absinthium Pakistan Aerial Aqueous- In vivo CCl4 and Gilani and
L. parts methanol Aceta- Janbaz (1995a)
minophen
Artemisia maritima L Pakistan Aerial Aqueous- In vivo CCl4 and Janbaz and Gilani
parts methanol Aceta- (1995)
minophen
Artemisia vulgaris L. Pakistan Aerial Aqueous- In vivo GAIN and LPS Gilani et al.
parts methanol (2005b)
Bidens chilensis DC Taiwan Whole Methanol In vivo CCl4 and PCM Chih et al.
plant (1996)
Bidens pilosa L. Taiwan Whole Methanol In vivo CCl4 and PCM Chih et al. (1996)
plant
Cichorium intybus L. India Seeds Alcohol In vivo CCl4 Ahmed et al.
(2003)
Crassocephalum Japan Whole Aqueous In vivo GAIN, LPS Aniya et al.
crepidioides Benth plant and in and CCl4 (2005)
vitro
Elephantopus mollis Taiwan Whole Aqueous In vivo Acetaminophen Lin et al.
Kunth. plant and GAIN (1995b)
Elephantopus Taiwan Whole Aqueous In vivo Acetaminophen Lin et al. (1995b)
scaber L. plant and GAIN
Flaveria trinervia India Leaf Methanol In vivo CCl4 Umadevi et al.
(Spreng.) C.Mohr (2004)
Gundelia Iran Stalk Hydro- In vivo CCl4 Jamshidzadeh
tourenfortii L. alcoholic and in et al. (2005)
vitro
Pseudoelephantopus Taiwan Whole Aqueous In vivo Acetaminophen Lin et al. (1995b)
spicatus (Juss. Ex plant and GAIN
Aublet) Gleason
Wedelia chinensis Taiwan Whole Methanol In vivo CCl4, Aceta- Lin et al. (1994)
(Osbeck) Merr. plant minophen and
GAIN
Wedelia India Leaf Ethanol In vivo CCl4 Murugaian et al.
calendulacea L. (2008)
Balanophoraceae Ghana Roots, Aqueous In vivo GAIN and Gyamfi et al.
Thonningia leaves and in CCl4 (1999)
sanguinea Vahl. vitro
Bixaceae Mali Rhizome Ethanol In vivo CCl4 Diallo et al.
Cochlospermum and hydro- (1992)
tinctorium Perri ethanol
ex Rich. extract
Bixa orellana L. Bangla- Seed Methanol In vivo CCl4 Ahsan et al.
desh (2009)
Brassicaceae India Whole plant Aqueous In vivo CCl4 Mantena
Coronopus didymus L. et al. (2005)
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
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72Hippocratic Journal of Unani Medicine
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
Burseraceae Saudi Aerial Ethanol In vivo CCl4 Al-Howiriny et al.
Commiphora Arabia parts (2004)
opobalsamum (L.)
Engl.
Caesalpiniaceae India Bark Methanol In vivo CCl4 and PCM Gupta et al.
Bauhinia racemosa (2004)
Lam
Capparidaceae India Leaves Ethanol In vivo CCl4 Gupta et al.
Cleome viscosa L (2009)
Casuarinaceae Bangla- Leaves, Methanol In vivo CCl4 Ahsan et al.
Casuarina desh bark (2009)
equisetifolia Forst
Celasteraceae Salacia Sri Lanka, Root, Aqueous, In vivo CCl4 Yoshikawa et al.
reticulata Wight India stem methanol (2002)
Chenopodiaceae Beta India Root Ethanol In vivo CCl4 Agarwal et al.
vulgaris L. (2006)
Combretaceae Japan Leaves Methanol In vivo GAIN Banskota et al.
Combretum Kurz. and in (2003)
vitro
Terminalia arjuna L. India Bark Aqueous In vivo CCl4 Manna et al.
(2006)
Terminalia belerica India Fruits Ethanol In vivo CCl4 Jadon et al.
Roxb (2007)
Terminalia catappa L. Okinawa Leaves Aqueous In vivo GAIN and LPS Kinoshita et al.
Island and in (2007)
vitro
Terminalia chebula India Fruits Ethanol In vivo Anti TB drugs Tasduq et al.
Reiz. and in (2006)
vitro
Compositae Ambrosia Egypt Whole Aqueous- In vivo Acetaminophen Ahmed and
maritima L. plant methanol Kharter (2001)
Crepis rueppellii Yemen Whole Ethanol In vivo CCl4 Fleurentin et al.
(Sch.) Bip. plant (1986)
Eclipta alba Hassk. India Whole Alcohol In vivo CCl4 Singh et al.
plant (1993)
Epaltes divaricata India Whole Aqueous In vivo CCl4 Hewawasam et al.
(L.) Cav. plant (2004)
Convolvulaceae Korea Seeds Aqueous In vivo DMN Kim et al. (2007a)
Cuscutae semen Lam.
Erycibe expansa Thailand Stem Methanol In vitro GAIN Matsuda et al.
Wall. and G.Don (2004)
Crassulaceae India Leaves Juice of In vivo CCl4 Yadav and Dixit
Kalanchoe pinnata leaves, and (2003)
Pers. ethanol in vitro
extract of
marc
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73Hippocratic Journal of Unani Medicine
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
Cucurbitaceae Luffa India Fruits Pet.ether, In vivo CCl4 Ahmed et al.
echinata Roxb. acetone, (2002)
methanol
Cyperaceae Cyperos Indonesia, Tubers Aqueous- In vivo CCl4 Gilani and
scariosus R.Br. Pakistan methanol Jambaz (1995b)
Ebenaceae Diospyros India Bark Methanol In vivo CCl4 Mondal et al.
malabarica (Desr.) (2005)
Kostel
Euphorbiaceae Nigeria Leaves Ethanol In vivo Acetaminophen Olaleye et al.
Alchornea cordifolia (2006)
Schum and Thonn.
Croton oblongifolius India Aerial Pet.ether, In vivo CCl4 Ahmed et al.
Roxb. parts acetone, (2002)
methanol
Emblica officinalis India Fruits Hydro- In vitro Anti TB drugs Tasduq et al.
Gaertner alcoholic (2005)
Phyllanthus India Whole n-hexane In vivo CCl4 and Asha et al. (2007)
maderaspatensis L. plant Thioacetamide
Phyllanthus niruri L. Brazil Leaves Aqueous In vivo PCM Sabir and Rocha
(2008)
Phyllanthus India Aerial parts Ethanol In vivo CCl4 Das et al.
reticulatus Poir. (2008)
Fabaceae Acacia India Bark Ethyl acetate In vivo CCl4 Ray et al.
catechu (L.f.) Willd. (2006)
Bauhinia variegata L. India Stem Alcohol In vivo CCl4 Bodakhe and
bark Ram (2007)
Cajanus cajan L. India Leaves Methanol In vivo Alcohol Kundu et al.
(2008)
Cassia fistula L. India Leaves n-heptane In vivo CCl4 Bhakta et al.
(1999)
Cassia occidentalis L. India Leaves Aqueous- In vivo PCM and Jafri et al. (1999)
ethanol ethyl alcohol
Glycine max (L.) Merr Taiwan Seed Water In vivo Acetaminophen Wu et al. (2001)
Phaseolus aureus Taiwan Seed Water In vivo Acetaminophen Wu et al. (2001)
Roxb.
Phaseolus calcaratus Taiwan Seed Water In vivo Acetaminophen Wu et al. (2001)
Roxb
Phaseolus radiatus L. Taiwan Seed Water In vivo Acetaminophen Wu et al. (2001)
Pterocarpus India Stem Methanol In vivo CCl4 Mankani et al.
marsupium Roxb. bark (2005)
Trigonella foenum- India Leaves Ethanol H2O2 CCl4 Meera (2009)
graecum L.
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74Hippocratic Journal of Unani Medicine
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
Fumariaceae Fumaria India Whole Methanol, In vivo PCM, Rao and Mishra
indica (Hausskn.) plant Pet.Ether, Rifampicin, (1997)
Pugsley aqueous CCl4
Fumaria parviflora Pakistan Shoots Aqueous- In vivo PCM Gilani et al.
Lam. methanol (1996)
Gentianaceae India Whole Alcohol In vivo CCl4 Senthilkumar
Enicostemma littorale plant et al. (2005)
Blume.
Swertia japonica Japan Whole Butanol In vivo GAIN Hase et al.
(Roem. and Schult.) plant (1997b)
Makino.
Lamiaceae Ocimum India Leaves Ethanol H2O2 CCl4 Meera et al.
basilicum L. (2009)
Moraceae Ficus India Leaves Methanol In vivo CCl4 Krishna et al.
carica L. (2007)
Ficus hispida L India Leaves Methanol In vivo PCM Mandal et al.
(2000)
Moringaceae Moringa Malaysia Leaves Hydro- In vivo Acetaminophen Fakurazi et al.
oleifera L. alcoholic (2008)
Myrtaceae Careya India Stem Methanol In vivo CCl4 Sambath et al.
arborea Roxb. bark (2005)
Nyctaginaceae India Roots Aqueous In vivo Thioacetamide Rawat et al.
Boerhaavia diffusa L. (1997)
Nymphaceae India Flowers Alcohol In vivo CCl4 Bhandarkar and
Nymphaea stellata Khan (2004)
Willd.
Oleaceae Phillyrea Jordan Leaves Aqueous In vivo CCl4 Janakat and
latifolia L. Al-Merie (2002)
Ophioglossaceae India Rhizomes Methanol In vivo CCl4 Suja et al. (2004)
Helminthostachys
zeylanica (L.) Hook
Orchidaceae Taiwan Whole Aqueous In vivo CCl4 Wu et al. (2007)
Anoectochilus plant and
formosanus Hayata in vitro
Polygalaceae India Leaves Chloroform In vivo GAIN Dhanabal et al.
Polygala arvensis (2006)
Willd.
Rhamnaceae Taiwan Bark Methanol, In vivo CCl4 Lin et al. (1995a)
Ventilago leiocarpa ethanol,
Benth. butanol
and
aqueous
Ziziphus mauritiana Nigeria Leaves Ethanol In vivo CCl4 Dahiru et al.
Lam. (2005)
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75Hippocratic Journal of Unani Medicine
Family and Origin Plant Extracts Type of Hepato- References
Botanical name Parts Studied assay toxicity
used Inducing
Agents
Rubiaceae Hedyotis India Whole Methanol In vivo PCM Sadasivan et al.
corymbosa (L.)Lam. plant (2006)
Mitracarpus scaber Mali Whole Methanol In vivo CCl4 Germano et al.
Zucc. plant and (1999)
in vitro
Morinda citrifolia L. America Whole Methanol In vivo CCl4 Wang et al.
plant (2008)
Rutaceae Aegle India Leaves Fine In vivo Alcohol Singanan et al.
marmelos (L.) Corr. powder (2007)
Serr. inphysio-
logical
saline
Glycosmis Bangla- Leaves, Methanol In vivo CCl4 Ahsan et al.
pentaphylla Corr. desh bark (2009)
Scrophulariaceae India Whole Alcohol In vivo Morphine Sumathy et al.
Bacopa monniera (L.) plant (2001)
Pennell
Picrorrhiza kurroa Himalayas Rhizome, Ethanol In vivo GAIN Anandan and
(Roule.) Sans roots Devaki (1999)
Smilacaceae Smilax Saudi Roots Ethanol In vivo CCl4 Rafatullah et al.
regelii Killip and Arabia (1991)
Morton
Solanaceae Jordan Leaves, Aqueous In vivo CCl4 Janakat and
Nicotiana glauca flowers Al-Merie (2002)
Graham.
Solanum nigrum L India Fruits Ethanol In vivo CCl4 Raju et al. (2003)
Solanum Jerusalem Leaves Methanol In vivo CCl4 Vijayan et al.
pseudocapsicum and in (2003)
Hassl. vitro
Solanum trilobatum L. India Whole Methanol In vivo CCl4 Shahjahan et al.
plant (2004)
Umbelliferae Taiwan Leaves Aqueous In vitro Acetamino- Liu et al. (2006)
Bupleurum kaoi Liu phen and CCl4(Chao et Chuang)
Daucus carota L. Europe, Roots Aqueous In vivo CCl4 Bishayee et al.
Asia, (1995)
Africa
Foeniculum vulgare Turkey Seeds Essential In vivo CCl4 Ozbek et al.
Miller oi l (2003)
Valerianaceae India Rhizomes Ethanol In vivo CCl4 Ali et al. (2000)
Nardostachys
jatamansi D.C.
Vitaceae Rhoicissus South Roots Aqueous In vivo CCl4 Opoku et al.
tridentata(L.f.) Wild Africa (2007)
and R.B. Drumm
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76Hippocratic Journal of Unani Medicine
Table 2: Chemically defined Molecules with Hepatoprotective activity
Chemical substance Plant Plant part Class References
3,4-di-O- Lactuca indica L. Aerial parts Quinic acid Kim et al.
caffeoylquinic acid (2007b)
3,5-di-O-caffeoyl- Lactuca indica L. Aerial parts Quinic acid Kim et al.
muco-quinic acid (2007b)
5-O-(E)-p- Lactuca indica L. Aerial parts Quinic acid Kim et al.
coumaroylquinic acid (2007b)
α-Amyrin Protium heptaphyllum Trunk wood Triterpene Oliveira et al.
(Aubl.) March resin (2005)
β-Amyrin Protium heptaphyllum Trunk wood Triterpene Oliveira et al.
(Aubl.) March resin (2005)
Anastatin A Anastatica Whole plant Flavonoid Yoshikawa et al.
hierochuntica L. (2003)
Anastatin B Anastatica Whole plant Flavonoid Yoshikawa et al.
hierochuntica L. (2003)
18 β-glycyrrhetinic Glycyrrhiza uralensis Rhizomes Glycyrrhetinic Shim et al.
acid Fisch. acid (2000)
Tetrahydroswer- Swertia japonica Makino Aerial parts Xanthione Hase et al.
tianolin (1997b)
Gentiopicroside Swertia japonica Makino Aerial parts Iridoid Hase et al.
(1997b)
Sweroside Swertia japonica Makino Aerial parts Iridoid Hase et al.
(1997b)
Andrographolide Andrographis paniculata Aerial parts Diterpene Chander et al.
(Burm.f) Nees (1995)
Erycibenin A Erycibe expansa Stem Pterocarpane Matsuda et al.
(2004)
5,7,4´-trihydroxy-3´- Erycibe expansa Stem Isoflavone Matsuda et al.
Methoxyisoflavone (2004)
Genistein Erycibe expansa Wall. Stem Isoflavone Matsuda et al.
Ex G.Don. (2004)
Orobol Erycibe expansa Wall. Stem Isoflavone Matsuda et al.
Ex G. Don. (2004)
Mangiferin Salacia reticulata Abst. Roots Phenolic Yoshikawa et al.
compound (2002)
(-)-4´-O-methy- Salacia reticulata Abst. Roots Phenolic Yoshikawa et al.
lepigallocatechin compound (2002)
Thymoquinone Nigella sativa L. Aerial parts Quinone Daba and Abdel-
Rahman (1998)
Lithospermate B Salvia miltorhiza Bunge Roots Caffeic acid Hase et al.
(1997a)
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77Hippocratic Journal of Unani Medicine
Taxiresinol Enciostemma littorale Aerial parts Tetrahydro- Nguyen et al.
furan (2004)
(7´R)-7´- Enciostemma littorale Aerial parts Tetrahydro- Nguyen et al.
hydroxylariciresinol furan (2004)
Onitin Equisetum arvense L. Aerial parts Phenolic Oh et al. (2004)
compound
Luteolin Equisetum arvense L. Aerial parts Flavonoid Oh et al. (2004)
Quercetin-3-O-β-D- Saururos chinensis Aerial parts Flavonol Sung et al.
glucuronopyranoside (Lour.) Baill. glycoside (1997)
Quercetin-3-O-β-D- Saururus chinensis Aerial parts Flavonol Sung et al.
glucuronopyranosyl (Lour.) Baill. glycoside (1997)
methyl ester
Scropolioside-A Scrophularia koelzii Aerial parts Iridoid Garg et al.
Pennell glycoside (1994)
3-(S)-3-_- Goodyera Whole plant Aliphatic Du et al. (2000)
Dglucopyrano- schlechtendaliana glycoside
syloxybutanolide Reichb.G. matsumurana
Schltr.G. discolor
Kergawl
3-(S)-3-_-D- Goodyera schlechten- Whole plant Aliphatic Du et al. (2000)
glucopyranosyloxy- daliana Reichb.G. glycoside
4-hydroxybutanoic matsumurana Schltr.G.
acid discolor Kergawl.
Agathisflavone Canarium manii King Aerial parts Biflavonoid Anand et al.
(1992)
(S)-bakuchiol Psoralea corylifolia Aerial parts Monoter- Hyun et al.
Babchi penephenol (2001)
Monomethyl fumarate Fumaria indica Pugsley Whole plant Fumaric acid Rao and Mishra
(1998)
Wighteone Cudrania Roots Flavonoid Lin et al. (1996)
cochinchinensis (Lour.)
Kudo et Masam.
Naringenin Cudrania Roots Flavonoid Lin et al. (1996)
cochinchinensis (Lour.)
Kudo et Masam.
Torilin Cnidium monnieri (L.) Aerial parts Sesquiterpene Oh et al. (2002)
Cusson
Torilolone Cnidium monnieri (L.) Aerial parts Sesquiterpene Oh et al. (2002)
Cusson.
Allicin Allium sativum L. Cloves Allyl Vimal and Devaki
thiosulfinates (2004)
Kaempferol Rhodiola sachalinensis Roots Phenolic Song et al.
A.Bor. compound (2003)
Chemical substance Plant Plant part Class References
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78Hippocratic Journal of Unani Medicine
Chemical substance Plant Plant part Class References
Salidroside Rhodiola sachalinensis Roots Phenolic Song et al. (2003)
A.Bor. compound
1-O-galloyl-6-O- Combretum Seeds Gallic acid Adnyana et al.
(4-hydroxy-3,5- quadrangulare Kurz (2001)
dimethoxy)benzoyl-
β-d-glucose
Picroliv Picrorhiza kurroa Aerial parts Iridoid Visen et al.
Royle ex Benth. glycoside (1991)
Indigtone Indigofera tinctoria L. Aerial parts Aliphatic Singh et al.
nitrocompound (2001)
Acanthoic acid Acanthopanax Root bark Diterpene Park et al. (2004)
koreanum Nakai
Myristin Myristica fragrans Aerial parts Cetyl ester Morita et al.
Houtt. (2003)
Rutin Artemisia scoparia Aerial parts Flavonoid Janbaz et al.
Waldst. and Kit. (2002)
Troxerutin Artemisia scoparia Aerial parts Flavonoid Zhang et al.
Waldst. and Kit. (2009)
Neoandrographolide Andrographis paniculata Aerial parts Diterpene Chander et al.
(Burm.f.) Wall. Ex Nees (1995)
5-0-methyl-(E)- Acer mono Maxim. Leaves Stilbene Yang et al.
resveratrol.3-0-β-D glycoside (2005)
glucopyranoside
5-0-methyl-(E)- Acer mono Maxim. Leaves Stilbene Yang et al.
resveratrol.3-0- glycoside (2005)
_-Dapiofuranosyl-
1->6)-β-D
glucopyranoside
Corilagin Terminalia catappa L. Leaves Tannin Kinoshita et al.
(2007)
y-Amyrone Sedum sarmentosum Aerial parts Triterpene Amin et al.
Bunge (1998)
3-epi-y-amyrin Sedum sarmentosum Aerial parts Triterpene Amin et al.
Bunge (1998)
y-Amyrin Sedum sarmentosum Aerial parts Triterpene Amin et al.
Bunge (1998)
18β-hydroperoxy- Sedum sarmentosum Aerial parts Triterpene Amin et al.
olean.12-en-3-one Bunge (1998)
Rubiadin Rubia cordifolia L. Roots Anthraquinone Rao et al. (2006)
3,4,5-trihydroxy- Terminalia belerica Fruit Gallic acid Jadon et al.
benzoic acid Roxb. (2007)
Kinsenoside Anoectochilus Whole plant Furanone Wu et al. (2007)
formosanus Hay.
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79Hippocratic Journal of Unani Medicine
Discussion and Conclusion
The liver is the most important organ in the body. It has a pivotal role in
regulation of physiological processes. It is involved in several vital functions
such as metabolism, secretion and storage. Liver diseases are among the
most serious ailments. They may be classified as acute or chronic hepatitis
(inflammatory liver diseases), hepatosis (non inflammatory diseases) and
cirrhosis (degenerative disorder resulting in fibrosis of the liver).
Modern society has innate knowledge about the herbal treatment of liver disease
from many cultures. Research into plants traditionally used in the treatment of
liver disease has significantly advanced in the past 15 years, and much of
what has been discovered supports traditional knowledge.
Considering the enormous biodiversity resources over the world’s traditional
system and the high incidence of liver complications, the present review
extensively focuses on collection of data for different plants,
which are available in India and all over the world. These medicinal plants claimed
as liver protective agents are classified according to their biological source,
phytoconstituents; part used and plants in formulations. People from India are
still dependent on conventional therapies to treat liver complications. Because
of their easy availability and low cost. Since large mass of populations used
preferable herbal preparation, therefore there is need to be evaluate for their
proportion, their dose and rational behind combination in different polyhedral
preparation.
These herbal drugs have shown the ability to maintain the normal functional
statues of the liver with or without fewer side effects. These are the reason
that’s why herbal hepatoprotectives are mostly preferred by medical
practitioners.
It has been seen that herbal hepatoprotective drugs have less side effect or
interaction as compared to synthetic medicine but in other hand scientific
evidence from tests done to evaluate the safety and effectiveness of traditional
hepatoprotective medicine products and practices is limited and further study
of products and practices is needed.
Pharmacokinetic and toxicity studies have not disclosed any issues that could
limit the therapeutic use of these drugs. Also the study is required to identify
glycosides, flavonoids, triterpenes and phenolic compounds as classes of
compounds with hepatoprotective activity.
Further studies including clinical trials need to be carried out to ascertain the
safety of these compounds as a good alternative to conventional drugs in the
treatment of liver diseases
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80Hippocratic Journal of Unani Medicine
Since the traditional system of medicine recommends various hetptoprotective
agents and preparations to treat hepatic disorders. The management of lives
disease is still challenges to modern medicine. The modern allopathic drugs
have very little to offer for alleviation of hepatic ailments and some these drugs
adversely affect the liver function. A phytotherapeutic approach to modern
drug development provides many invaluable drugs from traditional medicinal
plants. Search for pure phytochemical as drug is time consuming and extensive.
Numerous plants and polyherbal formulations are being used for the treatment
of liver diseases.
Today, unfortunately the herbal resources have declined rapidly because more
than 80% of our total medicinal plants used by Indian pharmaceutical industry
are collected from their wild sources and they are not being grown or
domesticated so far. To meet their burgeoning demand is the need of the day.
Moreover, our natural resource base of medicinal plants is being depleting day
by day. Hence there is an urgent need to encourage field scale cultivation of
prioritized medicinal plants through government initiatives before it’s too late.
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91Hippocratic Journal of Unani Medicine
TAbstract
his study was carried out to detect the elevated serum Anti-
streptolysin O (ASO) in patients which were coming in OPD of Regional Research
Institute of Unani Medicine with various clinical conditions during the period of
January 2011 to December 2012. The serum samples were tested for ASO
antibodies by latex agglutination test. Total 205 patients including 30.24 %
male and 69.7 % female were tested for ASO serum levels. 60 were found
positive and 145 were negative. In 60 positive cases 16 were male and 44
were female. The prevalence of ASO antibody in total cases was 29.26 %. The
prevalence in male was 25.8 % and in female was 30.76 %. The presence of
elevated streptococcal antibody titers in such a population probably reflects a
medium background prevalence of streptococcal infections.
Keywords: Anti-streptolysin O, Agglutination, Streptococcal infections, Serum
Introduction
Streptococcus pyogenes (Group A Streptococcus/GAS) is one of the most
common and ubiquitous human pathogens. It causes a wide array of infections,
the most frequent of which are acute pharyngitis (‘‘strep throat’’) and impetigo
(pyoderma). It is also associated with two main non-suppurative sequelae:
acute rheumatic fever (ARF) and acute glomerulonephritis (AGN) (Bisno, 1991).
Rheumatic fever causes inflammation of tissues and organs and can result in
serious damage to the heart valves, joints, central nervous system and skin.
Susceptibility to rheumatic fever in certain individuals has been ascribed to a
number of factors. These include genetic determinants, for example, HLAs and
the presence of certain markers such as the B-cell alloantigens. One of the
factors originally considered in rheumatic fever susceptibility is an innate state
of immune hyperresponsiveness, particularly to streptococcal antigens (Meiselas
et al., 1961; Stollerman, 1972; Rejholec, 1957).
The detection of streptococcal infection is done by positive culture for group
A Streptococcus from the throat or through anti-streptococcal antibody tests
(Homer and Shulman, 1991). The throat cultures are spontaneously negative
due to previous antibiotic treatment, or positive due to a carriage state. Hence
streptococcal antibody tests have become a method commonly used to provide
evidence of preceding streptococcal infection (Fink, 1991).
During infection, the host may produce antibodies to one or more extracellular
products of group A streptococci, and these antibodies are useful markers of
recent streptococcal infection. Tests that measure antibodies to the extra-
Prevalence ofAnti-streptolysinO antibodies atBhadrakRegion, India
*Kishore Kumar,
Subhan A. Khan,
Mahe Alam,
Hakimuddin Khan,
Chander Pal,
Qamar Uddin,
Mukesh Kumar
and
L. Samiulla
Regional Research Institute of
Unani Medicine (CCRUM),
Mathasahi, Bhadrak-756100,
Odisha
July - September 2014, Vol. 9 No. 3, Pages 91-97
*Author for correspondence
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92Hippocratic Journal of Unani Medicine
cellular products are commercially available and more commonly used. The
anti-streptolysin O (ASO) assay was the first such test to be developed and
is still widely used. ASO is not only useful in the diagnosis of streptococcal
infections or complications, but also in the follow-up process and in evaluating
the effectiveness of treatments. It measures the ability of serum to neutralize
streptolysin O.
Material and Methods
Serum Collection
Serum samples were collected from 205 patients between the periods of January
2011 to December 2012 at Regional Research Institute of Unani Medicine in
Bhadrak, India. Blood samples from patients were obtained using a standard
2-mL syringe. Samples were allowed to coagulate for 30 minutes at room
temperature in a serum-collecting tube. The sample was centrifuged at 3000
rpm for 5 minutes. The serum was then separated by using micropipette.
Procedure
All the serum samples were tested by ASO kit (Span Diagnostics P. Ltd. India).
The instructions, reagents and accessories to follow were supplied with the kit.
Test serum and reagents were kept at room temperature before testing. 40ìl
patient’s serum within the circled area was placed on the clean and dry special
glass slide provided in the kit. One drop of well mixed ASO latex reagents was
added to serum. The reagent and serum using the applicator stick were mixed.
The slide was rotated and observed for agglutination macroscopically within
two minutes.
Results
Total 205 patients were included in this study. From them, 30.24 % male and
69.7 % female tested for ASO serum levels (Figure 1), 60 were positive and
145 were negative.
In 60 positive cases 16 were male and 44 were female. In 145 negative cases
46 were male and 99 were female (Figure 2). Highest positive case (27 patients)
was found in the age group of 21- 40 while 23 patients were positive in age
group of 41-60 (Figure 3). The prevalence of total case was 29.26 %. The
prevalence of total female was 30.76 % and the prevalence of total male was
25.8 % (Table 1).
The number of positive cases in different age group is given in table 2. The
highest prevalence of male among positive cases found in the age group 41-
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93Hippocratic Journal of Unani Medicine
Figure 2: Sex wise distribution of ASO Positive and ASO Negative cases at RRIUM,
Bhadrak
Figure 1: Blood sample collection in General OPD attendance at RRIUM, Bhadrak
Table 1: Sex wise prevalence of all streptococcal infection among General OPD at
RRIUM, Bhadrak
Sex ASO Positive ASO Negative Total (%) Prevalence Rate
(%)
Male 16 46 62 (30.24) 25.84
Female 44 99 143 (69.75) 30.76
Total 60 145 205 (100) 29.6
60 (6/16, 37.5%) and the highest prevalence of female among positive cases
found in the age group 21 – 40 (22/44, 50 %) (Figure 4, Figure 5). The
prevalence rate of male and female in age group 21-40 was 31.25 % and 50
% respectively. This indicates that the prevalence rate is higher in females
then male in this age group.
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94Hippocratic Journal of Unani Medicine
Figure 4: The comparative chart of ASO positive cases between Male and Female
in different age group
Figure 3: Age wise distribution of total Anti- streptolysin O (ASO) positive cases
The prevalence rate of male and female in age group 41-60 was 37.5 % and
38.6 % respectively which was almost similar. The overall data clearly indicates
that the prevalence of ASO positive case is slightly higher in females than
males.
Discussion
The serological test for ASO is commonly used to aid in the diagnosis of post-
streptococcal non-suppurative sequelae such as ARF and glomerulonephritis
(Kimoto et al., 2005; Batsford et al., 2002).
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95Hippocratic Journal of Unani Medicine
Conventional laboratory practice is to measure levels of antibodies to various
combinations of the extra-cellular Group A Streptococcus antigens. However,
Blyth and Robertson (2006) showed that the addition of anti-streptokinase
antibodies measurement did not increase the sensitivity and specificity of
serological testing for the diagnosis of acute post-streptococcal disease (Blyth
and Robertson, 2006). ASO test method is based on an immunologic reaction
between streptococcal exotoxins bound to biologically inert latex particles and
streptococcal antibodies in the test sample. Visible agglutination occurs when
increased antibody level, are present in the test specimen. A positive ASO titer
indicates nonspecific immune stimulation due to past streptococcal exposure
resulting in polyclonal gammopathy (Sainani and Sainani, 2006).
Fujikawa and Okuni (1979) observed that ASO elevation occurs only in 60%
of rheumatic fever. But after one more test is added either
Table 2: The prevalence rate of ASO among positive cases in different age group
Age group Total No. of No. of Positive No. of Positive
Positive cases Male (%) Female (%)
0-20 6 4 (25) 2 (4.5)
21-40 27 5 (31.25) 22 (50)
41-60 23 6 (37.5) 17 (38.6)
61-80 4 1 (6.25) 3 (6.8)
Total 60 16 (100) 44 (100)
Figure 5: The prevalence rate of ASO among positive cases in different age group
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96Hippocratic Journal of Unani Medicine
antideoxyribonuclease-B titre or streptokinase test, they were able to diagnose
rheumatic fever with 95% accuracy (Fujikawa and Okuni, 1979). In this study,
60 samples were positive out of 205 samples. Our study showed 29.26 %
prevalence of ASO positive cases whereas the same study which was conducted
in Nepal showed 45.45 % prevalence of ASO positive cases (Khan et al.,
2012). Similar study was also performed where 20.89 % prevalence was
observed (Kandel et al., 2007). Further in an another study conducted at CMS
Teaching Hospital, Bharatpur, Nepal, 4230 serum samples of rheumatic fever,
glomerulonephritis, rheumatic heart disease and rheumatic arthritis were tested
for ASO antibodies by latex agglutination test during the period of January
2003 to December 2009. Among them 1944 samples were positive and 2286
samples were found negative with overall prevalence of 45.95 %. (Dewasy et
al., 2010)
This clearly indicates that the ASO levels vary with age group of the study
population and geographical distribution. This study, together with data derived
from the present study indicates that the levels of the streptococcal antibodies
in healthy populations can vary substantially, depending on the frequency of
streptococcal infections in those populations. The presence of elevated
streptococcal antibody titers in such a population probably reflects a medium
background prevalence of streptococcal infections.
Acknowledgement
The authors are thankful to the Director General, Central Council for Research
in Unani Medicine, New Delhi for his cooperation and taking keen interest in
this work. We also thank to all staff of Biochemistry laboratory of RRIUM,
Bhadrak, for helping in biochemical investigations.
References
Batsford, S., Brundiers, M., Horbach, E. and Monting, J., 2002. Streptococcal
Cysteine Proteinase as a Seromarker of group A Streptococcal
(Streptococcus pyogenes) Infections. Scand J. Infect. Dis. 34: 407–12
Bisno, A. L., 1991. Group A Streptococcal Infections and Acute Rheumatic
Fever. N. Engl. J. Med. 325: 783–93
Blyth, C. C. and Robertson, P.W., 2006. Streptococcal anti-bodies in the
Diagnosis of Acute and post-streptococcal disease: streptokinase versus
streptolysin O and deoxyribonuclease B. Pathology. 38: 152–6.
Dewasy, B.L., Singh, Y .I., Jha, B.K. and Kapil, J., 2010. A rapid screening test
by the latex antistreptolysin O test for streptococcal infections in CMS
Teaching Hospital, Bharatpur, Nepal. Journal of college of Medical Sciences-
Nepal 6: 24-28
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97Hippocratic Journal of Unani Medicine
Fink, C. W., 1991. The Role of the Streptococcus in Post-streptococcal Reactive
Childhood Polyarteritis Nodosa. J. Rheumatol. Suppl. 29: 14–20.
Fujikawa, S. and Okuni, M., 1979. The determination of ADNase-B titres Jpn.
Circ. J. 43: 417–418.
Homer, C. and Shulman, S. T., 1991. Clinical aspects of acute rheumatic fever
J. Rheumatol. Suppl. 29: 2–13.
Kandel, N. P., Koirala, B., Shrestha, S., Kumar, R. and Basnyat, S. R., 2007.
Laboratory Tests for Infective Endocarditis among Patients Visiting Shahid
Gangalal National Heart Centre, Bansbari, Kathmandu, Nepal. Journal of
Nepal Health Research Council 5: 1-4.
Khan, S., Singh, P. and Siddiqui, A. H., 2012. Prevalence of anti-streptolysin
O antibodies at Banke region Nepal. Bali Medical Journal 1(3): 98-100.
Kimoto, H., Fujii, Y., Hirano, S., Yokota, Y. and Taketo, A., 2005. Expression of
Recombinant Streptolysin O and Speci Production. J. Mol. Microbiol.
Biotechnol. 10: 64–68.
Meiselas, L.E., Zinglae, S.B. and Lee, S.L. 1961. Antibody production in rheumatic
diseases, the effect of brucella antigen. J. Clin. Investig. 40: 1872-1881.
Rejholec, V., 1957. Incidence of rheumatic fever in relation to immunologic
reactivity. Ann Rheum. Dis. 16: 23-30.
Sainani, G. S. and Sainani, A. R., 2006. Rheumatic Fever - How Relevant in
India Today? Journal of the Association of Physicians of India 54: 42-47.
Stollerman, G.H., 1972. Hypersensitivity and antibody responses in streptococcal
disease. In: L.M. Wannamaker and J.M. Matesen (Eds.) Streptococci and
streptococcal diseases: recognition, understanding and management.
Academic Press, New York, pp. 501-513.
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98Hippocratic Journal of Unani Medicine
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99Hippocratic Journal of Unani Medicine
TAbstract
he dried mature roots of Withania somnifera (L.) Dunal are
specified source of drug named ‘Asgand’ or ‘Ashwagandha’. The drug is highly
regarded for varied therapeutic values in Unani, Ayurvedic, Siddha and
Homoeopathic system of medicine. The drug has been compared to Panax
ginseng for its endurance - enhancing properties. The drug enjoys the official
status in Unani Pharmacopoeia of India, Ayurvedic Pharmacopoeia of India,
Siddha Pharmacopoeia of India, Homoeopathic Pharmacopoeia of India and
Indian Pharmacopoeia. Pharmacopoeial monographs are regulatory standards
and mandatory for compliance to ensure the quality, safety and efficacy of
drug. This communication reviews the pharmacopoeial monographs and
analytical parameters specified in pharmacopoeial and non pharmacopoeial
publications with a view on harmonization of quality specifications on W.
somnifera of drug in pharmacopoeias.
Keywords: Withania somnifera (L.) Dunal, Pharmacopoeias, Pharmacopoeial
harmonization.
Introduction
W. somnifera (L.) Dunal. (Family-Solanaceae) is specified botanical source of
‘Asgand’ or ‘Ashwagandha’ in various pharmacopoeias. The drug has been
used in Unani system of medicine since a long time. It is an official drug and
one of the ingredient of a number of important Unani classical and patent
formulations. The Ashwgandha is also used in ayurvedic, siddha, homoeopathic
and modern system of medicine. W. somnifera (L.) Dunal. is attributed medicinal
as aborfacient, alexipharmica, alternative, aphrodisiac, astringent, deobstruent,
diuretic, hypnotic and sedative, narcotic, pungent, restorative and tonic. It is
medicinally used in carbuncles, cough, debility of old age, dropsy, emaciation
of children, general weakness, promotes urination, functional obstruction of
body, rheumatism, scrofula, senile decay, ulcers and vulnerary treatment. The
drug finds mention in classical literature of unani, ayurvedic, siddha and
homoeopathic system of medicine. The tender shoots of the plant are also
used as a vegetable and seeds are used as masticatory. The green berries
are bruised and rubbed on ringworm in human beings and on animal sores
and girth-golls in horses. They are also employed to curdle milk. The drug is
substituted with W. coagulans Dunal. (Kirtikar and Basu, 1933; Chopra et al.,
1956; Nadkarni, 1954; Anonymous, 1976).
Review onPharmacopoeialStandards of‘Asgand’(Withaniasomnifera (L.)Dunal)
1*Nitin Rai
and2Rajeev Kr. Sharma
1Pharmacopoeial Laboratory
for Indian Medicine,
Kamla Nehru Nagar,
Ghaziabad-201002
2Pharmacopoeia Commission
for Indian Medicine,
Raj Nagar, Ghaziabad-201002
July - September 2014, Vol. 9 No. 3, Pages 99-109
1*Author for correspondence
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100Hippocratic Journal of Unani Medicine
Systematics of source plant : W. somnifera (L.) Dunal belongs to family
Solanace. Solanaceae Juss. Lindl. Veg. Kingd. 618, Endl. Gen. 662. Miers,
Illustr. S. Amer. Pl. 1849-1857, Gen. Pl. II: 882.
This family comprises about 20 genera and 2,000 species found in tropical
and temperate regions of the world with the prime centers in Central and South
America. In India, this family is spread over 15 genera and 88 species, mostly
in the Himalayas and the mountains of Southern India.
Genus: Withania Pauquy. Diss. de Bellad. Paris, 1824, ex Endl. Gen. 666;
Gen. Pl. II: 893, FBI 4:239.The genus consists of ten species which are
distributed in South America, South Africa, Canaries and Mediterranean to
India. In India, 2 species are distributed in drier regions. The generic name of
the plant is supposed to be in honour of H. Withan, a British geologist and
writer on Paleontobotany, in the nineteenth century.
W. somnifera (L.) Dunal. In DC. Prodr. 13: 453. 1852; FBI 4:239; FUGP 2:128,
Physalis somnifera Linn. Sp. Pl. 182. 1753, P. flexuosa Linn. Sp. Pl. 182. 1753.
Synonyms: Physalis flexuosa Linn and P. somnifera Linn. An erect much
branched, evergreen, perennial, undershrub with glutinous cells sap. Roots
fairly long, stem very short, stellate hairy often with hoary mealy exteremites.
Leaves simple, short petioled, alternate, or in sub-opposite pairs at a node,
exstipulate, elliptic-oblong or ovate rounded with an acute decurrent base.
Flowers greenish or yellowish, small, sessile or short pedicellate, axillary fascicles
or in crowded fascicles of two to five. Calyx gamosepelous, five or six lobed,
campanulate, stellate-hairy, lobes ovate-triangular, acute, corolla small,
gamopetalous, campanulate, three to six lobed to more than half way down,
ovate, stellate, hairy outside. Stamens five, inserted on the tube of the corolla
near its base, anthers broadly elliptic oblong and dehiscing longitudinally, pistil
bicarpellary, many ovuled, syncarpous, ovary ovate-globose, style glabrous,
linear filiform and stigma bifid. Fruit berry, globose, enclosed within the enlarged
calyx, stellate hairy, seed very many and discoid. Flowering and Fruiting:
January to September.
Distribution : The drug plant is distributed throughout the drier subtropical
regions of India. In global distribution it is reported from Arabia, Mediterranean
regions, Pakistan to Persia, the Canaries and to tropical and South Africa
(Chopra et al., 1958; Sharma and Kachroo, 1983).
Observations
The Pharmacopoeial monographs on herbal drugs published in Unani, Ayurveda,
Siddha, Homeopathic and Indian Pharmacopoeias are dealt in detail for content
review (Rai et al., 2012, 2013 and Tiwari et al., 2013).It was observed that
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101Hippocratic Journal of Unani Medicine
drugs ‘Asgand’ or ‘Ashwagandha’ (botanically specified as W. somnifera (L.)
Dunal. ) is subjected for regulatory standards in Unani Pharmacopoeia of
India, Ayurvedic Pharmacopoeia of India, Siddha Pharmacopoeia of India,
Homeopathic Pharmacopoeia of India, Indian Pharmacopoeia, British
Pharmacopoeia and United State Pharmacopoeia (Table -1). Non regulatory
quality standard are also published on the drugs in Indian Herbal Pharmacopoeia
and Quality Standards of Indian Medicinal Plants. (Anonymous, 1940; 1955;
1966; 1971; 1976; 1986; 1996;1998; 2000a & b; 2002; 2007; 2008 a & b;
2010; 2011; 2013 a & b and 2014.) The monographs published in these
pharmacopoeial and non-pharmacopoeial publications are reviewed as per the
pharmacopoeial parameters to assess the variability in quality standards of
drug.Table-2 exhibit the format of pharmacopoeial monographs in various edition
of pharmacopoeias.Table-3 & 4 provide the account of pharmacopoeial
standards on drug published in pharmacopoeias. Non- regulatory quality
standard published in Indian Herbal Pharmacopoeia and Quality Standards of
Indian Medicinal Plants are enumerated in Table-5.
Conclusion
India is the only country which recognizes the five pharmacopoeias of different
systems of medicines under regulatory frame work (Anonymous, 1940). Indian
Table 1: Pharmacopoeial status of Withania somnifera (L.) Dunal.
Botanical Name Morphological Pharmacopoeial/ Regulatory/
(as specified in Part specified Monograph Title Pharmacopoeial
Pharmacopoeial as drug References
Monograph)
Withania somnifera Dried mature Ashwagandha IP 2007 IP 2010 &
(L.) Dunal root IP 2014
Ashwagandha IP 2010 & IP 2014
dry extract
Amukkara SPI-I
Withania radix, IP 55& IP 66
Aswagandha
Asvagandha API- I & VIII
Asgand UPI-I
Withania somnifera HPI-I & VIII
Withania Somnifera British Pharmaco-
Root poeia (BP) 2013
Asvagandha Root The United States
Pharmacopoeia
(USP 36/NF 31) 2013
Abbreviations: API-Ayurvedic Pharmacopoeia of India, UPI-Unani Pharmacopoeia of India, SPI-
Siddha Pharmacopoeia of India, HPI-Homeopathic Pharmacopoeia of India, IP-Indian
Pharmacopoeia, BP -British Pharmacopoeia, USP-United States Pharmacopoeia India.
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102Hippocratic Journal of Unani Medicine
Table 2: Monograph pattern in various edition of Indian Pharmacopoeia.
Sl. Quality Unani Ayurvedic Siddha Homoeopathy Indian
No. Specification Pharmaco- Pharmaco- Pharmaco- Pharmacopoeia Pharmacopeia
poeia of poeia of poeia of of India (IP 2014)
India, India, India, (HPI)
Part–I Part-I Part-I Volume-I
(UPI) (API) (SPI) & VIII
Volume-I Volume-I Volume-I
& VIII
1. Pharmacopoeial Title √ √ √ √ √
2. Definition-Botanical √ √ √ Botanical √Name (family), Name, Family,
Part used as Part used,
distribution Distribution
are under
independent
headings
3. Synonyms √ √ √ √ √
4. Regional Language √ √ √ Common Names –
Name
5. Description √ √ √ Description- –
Macroscopic Macroscopic
Microscopic Microscopic
Powder Powder-
independent
headings
6. Identity, Purity & √ √ √ – –
Strength
Foreign Matter √ √ √ – √
Total Ash √ √ √ – √
Acid insoluble ash √ √ √ – √
Alcohol/ethanol √ √ √ – √soluble extractive
Water soluble √ √ √ – √Extractive
Loss on drying – √ – – √
Heavy metals – √ – – √
Microbial – √ – – √contamination
Pesticide residues – √ – – –
Aflatoxins – √ – – –
7. Assay √ √ – – √
8. Thin Layer √ √ √ In certain √Chromatography monographs
9. Constituents √ √ √ – –
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103Hippocratic Journal of Unani Medicine
10. Properties and √ √ √ – Category
Action (as per
system of
medicine)
11. Important √ √ √ – –
Formulations
12. Therapeutic Uses √ √ √ – –
13. Dose √ √ √ – –
14. Identification – – – √ Macroscopic
Microscopic
& TLC
15. History and authority – – – √ –
16. Preparation – – – √ –
17. Heavy metals – – – √ √
18. Loss on drying – – – √ √
19. Microbial – – – √ √contamination
20. Storage – – – √ √
Sl. Quality Unani Ayurvedic Siddha Homoeopathy Indian
No. Specification Pharmaco- Pharmaco- Pharmaco- Pharmacopoeia Pharmacopeia
poeia of poeia of poeia of of India (IP 2014)
India, India, India, (HPI)
Part–I Part-I Part-I Volume-I
(UPI) (API) (SPI) & VIII
Volume-I Volume-I Volume-I
& VIII
Table 3: Comparative account of pharmacopoeial standards on Withania somnifera (L.)
Dunal published various edition of Indian Pharmacopoeia.
Sl. Quality Unani Ayurvedic Siddha Homoeopathy Indian
No. Specification Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopeia
of India (UPI), of India (API), of India (SPI), of India (HPI) (IP 2014)
Part-I, Part-I, Part-I, Volume-I & VIII
Volume-I Volume-I & VIII Volume-I
1. Official Title Asgand Asvagandha Amukkara Withania Ashwagandha
somnifera Indian Ginseng;
(With. Som.) Withania
somnifera
2. Botanical Withania Withania Withania Withania Withania
Species somnifera Dunal. somnifera (L.) somnifera (L.) somnifera somnifera (L.)
Dunal. Dunal. Dunal. Dunal.
3. Synonyms – – Physalis Physalis –
somnifera L., somnifera L.,
P. flexuosa L., P. flexuosa L.
P. arborescens
DC.
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104Hippocratic Journal of Unani Medicine
4. Morphological Dried mature Dried mature Dried roots Root Root
part/Official roots roots
part
5. Description I. Macroscopic I. Macroscopic I. Macroscopic I. Macroscopic I. Macroscopic
II. Microscopic II. Microscopic II. Microscopic II. Microscopic II. Microscopic
III. Powder III. Powder
6. Identity, Purity
& Strength
Foreign Matter 2.0 %, Not 2.0 %, Not 2.0 %, Not – 2.0 %, Not
more than more than more than more than
Total Ash 7.0 %, Not 7.0 %, Not 7.0 %, Not – 7.0 %, Not
more than more than more than more than
Acid insoluble 1.0%, Not 1.0%, Not 1.0%, Not – 1.2%, Not
ash more than more than more than more than
Alcohol/ 15.0%, Not 15.0%, Not 15.0%, Not – 10.0%, Not
ethanol soluble less than* less than* less than less than
extractive (Vol. VIII)
Water soluble – 7.0%, Not 27.0%, Not – 15.0%, Not
Extractive less than less than less than
(Vol. VIII)
Loss on drying – 12.0 %, Not – – 12.0%, Not
more than more than
Heavy metals – Pharmacopoeial – – Pharmacopoeial
limits (Vol. VIII) limits
Microbial – Pharmacopoeial – – Pharmacopoeial
contamination limits (Vol. VIII) limits
Pesticide – Pharmacopoeial – – –
residues limits (Vol. VIII)
Aflatoxins – Pharmacopoeial – – –
limits (Vol. VIII)
7. Assay Total alkaloids- Total alkaloids- Assay by HPLC, – Total withanoloid-
0.2 % Not 0.2 % Not Quantification 0.2 % Not
less than less than not given less than
8. Chromato- TLC profile TLC/HPTLC TLC profile – TLC profile
graphy profile (Vol. VIII)
(TLC/HPTLC/
HPLC)
*Alcohol (25 percent) soluble extractive
Sl. Quality Unani Ayurvedic Siddha Homoeopathy Indian
No. Specification Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopeia
of India (UPI), of India (API), of India (SPI), of India (HPI) (IP 2014)
Part-I, Part-I, Part-I, Volume-I & VIII
Volume-I Volume-I & VIII Volume-I
Pharmacopoeia (IP) is the first official pharmacopoeia having its first edition in
the year 1955 followed by the publication of other pharmacopoeias viz. Ayurvedic
Pharmacopoeia of India (1986), Unani Pharmacopoeia of India (1998), Siddha
Pharmacopoeia of India (2008b) and Homoeopathic pharmacopoeia of India
(1971). All these Pharmacopoeias provide regulatory standards under Drugs
& Cosmetics Act 1940 & Rules thereunder for quality control of drugs of
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105Hippocratic Journal of Unani Medicine
Table 4: Comparative account of pharmacopoeial standards on Withania somnifera (L.)
Dunal published other Pharmacopoeias
Sl. Quality American Herbal British The United States
No. Specification Pharmacopoeia Pharmacopoeia Pharmacopoeia
(AHP) 2000 (BP) 2013 (USP 36/NF 31) 2013
1. Official Title Asvagandha Root Withania Somnifera Asvagandha Root
Root
2. Botanical Species Withania somnifera Withania somnifera Withania somnifera
(L.) Dunal. (L.) Dunal. (L.) Dunal.
3. Morphological part/ Dried root Dried mature root Dried mature root
Official part
4. Description I. Macroscopic I. Macroscopic I. Macroscopic
II. Microscopic II. Microscopic II. Microscopic
III. Powder III. Powder
5. Identity, Purity & Strength
Foreign Matter 2.0%, Not more than – 2.0%, Not more than
Total Ash 7.0%, Not more than 7.0%, Not more than 7.0%, Not more than
Acid insoluble ash 1.0%, Not more than 1.0%, Not more than 1.0%, Not more than
Alcohol/ethanol – – 10.0%, Not less than
soluble extractive
Water soluble – – –
Extractive
Loss on drying 10.0%, Not more than 12.0%, Not more than 12.0%, Not more than
Heavy metals - - Pharmacopoeial limits
Microbial Pharmacopoeial limits - Pharmacopoeial limits
contamination
Pesticide residues - - Pharmacopoeial limits
Aflatoxins - - Pharmacopoeial limits
6. Assay Total alkaloids-0.2% Withaferin A-0.1% Withanolides- 0.3%
Not less than Not less than and Not less than
Withanolide 0.1%
Not less than
7. Chromatography HPLLC profile TLC profile TLC profile
(TLC/HPTLC/HPLC)
Ayurvedic, Siddha, Unani, Homoeopathic and modern systems of medicine.
Pharmacopoeial standards on W. somnifera (L.) Dunal. in respect of total ash,
acid insoluble ash, alcohol/ethanol soluble extractives and water soluble
extractive varies in different pharmacopoeial monographs (Table-6). To facilitate
uniformity in regulatory quality specifications, harmonization of pharmacopoeial
standards is very much needed when acceptability of herbal drugs in uniform
manner is accelerating (Rai and Sharma, 2014).
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106Hippocratic Journal of Unani Medicine
Table 5: Comparative account of pharmacopoeial standards on Withania somnifera (L.)
Dunal published on non Pharmacopoeial.
Sl. Quality Indian Herbal Quality Standards of
No. Specification Pharmacopoeia Indian Medicinal Plants
(IHP) 2002 (QCIMP) 2011
1. Official Title Withania Somnifera Withania somnifera (L.)
Dunal.(Asvagandha)
2. Botanical Species Withania somnifera (L.) Withania somnifera (L.)
Dunal. Dunal.
3. Synonyms Physalis somnifera L., Physalis somnifera L.
P. flexuosa L.,
P. arborescens DC.
4. Morphological part/ Dried roots Dried roots
Official part
5. Description I. Macroscopic I. Macroscopic
II. Microscopic II. Microscopic
III. Powder
6. Identity, Purity & Strength
Foreign Matter 2.0%, Not more than 1.0%, Not more than
Total Ash 7.0%, Not more than 8.0%, Not more than
Acid insoluble ash 1.2%, Not more than 2.0%, Not more than
Alcohol/ethanol soluble 20.0%, Not less than 18.0%, Not less than
extractive 20.0%, Not less than*
Water soluble Extractive – 22.0%, Not less than
7. Assay Quantification not given Withaferine A - 0.036
to 0.094
8. Chromatography HPLC profile TLC/HPLC profile
(TLC/HPTLC/HPLC)
*Alcohol (25 percent) soluble matter.
Pharmacopoeial standards on W. somnifera (L.) Dunal. incorporated in various
pharmacopoeias are required to be harmonized with the monographs appeared
in other contentmpoarory pharmacopoeias. The quality of herbal drugs is
always prime issue and dealt with regulatory provisions of pharmacopoeial
monographs. The harmonized pharmacopoeial monographs will be yardstick to
ensure the quality, safety and efficacy of herbal drugs without any ambiguity.
References
Anonymous, 1940. Drugs & Cosmetics Act and Rules, Govt. of India, Ministry
of Health & Family Welfare, New Delhi.
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107Hippocratic Journal of Unani Medicine
Table 6: Variation in standards in respect of Pharmacopoeial standards.
Sl. Quality Unani Ayurvedic Siddha Homoeopathy Indian
No. Specification Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopoeia Pharmacopeia
of India (UPI), of India (API), of India (SPI), of India (HPI) (IP 2014)
Part-I, Part-I, Part-I, Volume-I & VIII
Volume-I Volume-I & VIII Volume-I
1. Foreign Matter 2.0%, Not 2.0%, Not 2.0%, Not – 2.0%, Not
more than more than more than more than
2. Total Ash 7.0%, Not 7.0%, Not 7.0%, Not – 7.0%, Not
more than more than more than more than
3. Acid insoluble 1.0%, Not 1.0%, Not 1.0%, Not – 1.2%, Not
ash more than more than more than more than
4. Alcohol/ 15.0%, Not 15.0%, Not 15.0%, Not – 10.0%, Not
ethanol soluble less than less than less than less than
extractive (Vol. VIII)
5. Water soluble – 7.0%, Not 27.0%, Not – 15.0%, Not
Extractive less than less than less than
(Vol. VIII)
6. Microbial – Pharmacopoeial – – Pharmacopoeial
contamination limits (Vol. VIII) limits
7. Pesticide – Pharmacopoeial – – –
residues limits (Vol. VIII)
8. Aflatoxins – Pharmacopoeial – – –
limits
(Vol. VIII)
9. Assay Total alkaloids- Total alkaloids- Quantification – Total
0.2% Not 0.2% Not not given withanoloid-
less than less than 0.2% Not
less than
Anonymous, 1955. Pharmacopoeia of India (The Indian Pharmacopoeia) first
ed., Govt. of India, Ministry of Health and Family Welfare, New Delhi.
Anonymous, 1966. Pharmacopoeia of India (The Indian Pharmacopoeia) second
ed., Govt. of India, Ministry of Health and Family Welfare, New Delhi.
Anonymous, 1971. Homoeopathic Pharmacopoeia of India Vol. I. Government
of India, Ministry of Health & Family Welfare, New Delhi.
Anonymous, 1976.The Wealth of India (Raw Materials), Vol. X (Sp-W). C.S.I.R.,
New Delhi.
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India, Ministry of Health & Family Welfare, New Delhi pp. 7-8.
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108Hippocratic Journal of Unani Medicine
Anonymous, 2000a. Homoeopathic Pharmacopoeia of India Vol. VIII. Government
of India, Ministry of Health & Family Welfare, New Delhi, pp.136-37.
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Indigenous Drugs of India. U.N. Dhur & Sons Pvt. Ltd., Calcutta.
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109Hippocratic Journal of Unani Medicine
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110Hippocratic Journal of Unani Medicine
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111Hippocratic Journal of Unani Medicine
TAbstract
he present study was carried out during 2011-2012 to
investigate the safety of Qurse-nazla by conducting acute and sub-acute toxicity
in Swiss albino mice & rats respectively. Acute toxicity was determined by
administering aqueous extract of Qurse-nazla orally to two groups of mice of
six each at a dose 1gm/kg and 2gm/kg body weight. The animals were observed
for gross behaviour and mortality for 24 hours after drug administration. The
formulation was well tolerated by the animals and no abnormality was observed
in the general behaviour (salivation, lacrymation, lethargy, sleep and coma) of
the animals and no overnight mortality was recorded. Similarly, sub-acute toxicity
was determined in albino rats by orally administration of aqueous extract to
three groups of seven animals each at a dose ranges from 1gm/kg and 3gm/
kg body weight for 28 days. The results of haematology and biochemistry
profile done on 29th day were found to be normal and no changes were
observed in organ to body wt ratio of liver, heart, kidney and spleen.
Keywords: Acute toxicity, Sub-acute toxicity, Qurse-Nazla, Ocimum sanctum
Linn., Cinnamomum zeylanicum Blume., Zingiber officinale Rose., Tinospora
cordifolia Wild.
Introduction
The World Health Organization is fully aware of the importance of herbal
medicines to the health of many people throughout the world, as stated in a
number of resolutions adopted by the World Health Assembly and the Regional
Committee for the Western Pacific. Thus herbal medicines have been recognized
as a valuable and readily available resource for primary health care, and WHO
has endorsed their safe and effective use. A comprehensive programme for
the identification, cultivation, preparation, evaluation, utilization and conservation
of herbal medicines has been developed. Meanwhile, it has been realized that
medicinal plants are a valuable resource for new pharmaceutical products and
thus a potential source of new drugs as well as for economic development
(Anonymous, 1993).
Determination of efficacy and safety of herbal remedies is necessary because
many people using these agents as self medication. Since there is limited data
available about the safety of the commonly used herbal remedies, therefore,
effort to elucidate health benefits and risk of herbal medicine should be
intensified. Toxicity testing in animal is typically the initial steps to determine the
effect profile of test substance and potential hazards which occurs due to short
term exposure of test substance. A toxicity study provides information on the
Acute & Sub-Acute ToxicityStudy of Qurse-Nazla inExperimentalAnimals
*Mohd. Nadeem,
Mohd. Urooj,
Habibur Rehman
and
Shariq Ali Khan
Pharmacology Research Unit
Regional Research Institute of
Unani Medicine (CCRUM),
Post Box 70, Aligarh-202002, U.P.
July - September 2014, Vol. 9 No. 3, Pages 111-118
*Author for correspondence
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112Hippocratic Journal of Unani Medicine
hazardous properties and allows the substance to be ranked and classified
according to the Globally Harmonized System (GHS) for the classification of
chemicals which cause acute toxicity (OECD, 2000).
Formulation
Qurse-nazla is a poly herbal formulation containing main constituents in equal
parts:
S.No. Common Name Botanical Name
1. Barg-e-Tulsi Ocimum sanctum Linn.
2. Darchini Cinnamomum zeylanicum Blume.
3. Zanjabeel Zingiber officinale Rose.
4. Satte Gilo Tinospora cordifolia Wild.
5. Satte Ajwain Ptychotica jowar DC
Methodology
This study was carried out in Pharmacology Research Unit, of Regional
Research Institute of Unani Medicine, Aligarh and conducted in accordance
with the protocol approved by Institutional Ethics Committee (RRIUM), Aligarh.
Procurement of Drug
The Qurse nazla formulation was procured in the form of Tablet from CRIUM
(Central research Institute of Unani Medicine) A.G colony Road Near ESI
Hospital, Eragadda Road, Hyderabad.
Animals
The study was carried out in Swiss albino mice (20-25 g) and rats (100-150g)
of either sex, for acute & sub-acute toxicity determination respectively. The
animals were procured from Mr. Rahat Hussain Enterprises Biological Suppliers
Babri Mandi Aligarh. They were acclimatized to the conditions for one week
before experimental study. The animals were maintained in a standard
environmental condition at a room temperature of (25±2 degree Celsius) with
12 Hrs light/Dark cycles, humidity (50-55%), and had free access to food
pellets. The study was conducted after approval of protocol from Institutional
ethics committee of RRIUM, Aligarh.
Preparation of Drug Extract
The tablets of the drugs were crushed into fine powder and a weighed quantity
was steeped in acidulated distilled water. The water soaked mass of the drug
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113Hippocratic Journal of Unani Medicine
was warmed over water bath and kept for 24 hours at room temperature.
During this period it was occasionally stirred. After 24 hours it was filtered
through a filter paper and filtrate was dried over water bath. The aqueous
extracts of the drug thus obtained was used in different doses selected according
to OECD guidelines for safety evaluations.
Acute Toxicity Study
The current study was carried out in accordance to Organization of Economic
Co-operation and Development (OECD) guideline for testing of chemicals.
Swiss Albino mice of either sex weighing 20-25 g were randomly selected and
divide into two groups of six mice each. Mice were kept fasted overnight
(12hrs) with free access to water prior to administration of dose ranging 1gm/
kg body weight and 2g/kg body weight as per limit test of OECD guideline. The
aqueous extract of the drugs was administered orally. The animals were kept
in polypropylene cages after drug administration and were observed for Gross
behaviour (salivation, lacrymation, lethargy, sleep and coma) & mortality at 1
hour, 2 hour, 3 hour, 4 hour, 5 hour, 6 hour, 12 hour, 24 hour and thereafter
once every day up to 14 days after drug administration.
Sub-Acute Toxicity Study
Swiss Albino rats of either sex weighing between 100-150 g were randomly
selected and divided into three groups of seven animals each. Rats were kept
fasted overnight (12hrs) with free access to water prior to administration of
dose ranging 1gm/kg and 3gm/kg body weight for 28 days as per limit test of
OECD guideline. Group I was kept as normal control which received distilled
water for 28 days, while in the IInd and IIIrd groups aqueous extract of the drug
was administered orally at a dose of 1gm/kg and 3gm/kg body weight for 28
days. The animals were observed for Gross behavior (salivation, lacrymation,
lethargy, sleep and coma) & mortality at 1 hour, 2 hour, 3 hour, 4 hour, 5 hour,
6 hour, 12 hour, 24 hour and thereafter once every day up to 28 days after
drug administration. On 29th day, blood was collected of all the three groups
of rats through retro-orbital plexus for estimation of SGOT,SGPT and Serum
alkaline Phosphatase, Serum urea and Serum Creatinine, Serum cholesterol,
Serum triglyceride and Serum HDL, Percentage hemoglobin, ESR, Total
leukocyte count and Differential leukocyte count. After collection of blood the
animals in all the three groups were sacrificed and liver, heart, kidney and
spleen were excised out for determination of organ to body weight ratio. SGOT,
SGPT were estimated by Reitman’s and Frankel, method (Reitman et al.,
1957). Alkaline Phosphatase was estimated by Bessey and Brock, 1946 method
(Bessay et al., 1946). Serum urea was estimated by GLDH, Ureas method
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114Hippocratic Journal of Unani Medicine
given by (Tiffany et al., 1972), while Serum Creatinine was estimated by Jaffe’s
method given by (Bower’s et al., 1980). Serum HDL was estimated by
Phosphotungstic Acid method given by (Burstein et al., 1970), while Serum
cholesterol and Triglyceride were estimated by CHOD-PAP method given by
(Roeschlau et al., 1974) and GPO-Trindev method given by (Mcgowan et al.,
1983). ESR and DLC were estimated by Westergreen and Leishman stain
methods given in Medical Laboratory Technology (Mukherjee, 1990). TLC was
estimated by Hemoaltometry method (Plum, 1936). Percent hemoglobin was
estimated by Sahli’s Acid Haematin method given by (Newcomer, 1919).
Statistical Analysis
Statistical analysis was performed by using unpaired t test calculating p value
at 5% level. All values are expressed as Mean±SEM (standard error of mean).
p value less than 0.05 found to be considered statistically significant.
Results and Discussion
Acute Toxicity Study
The effect of oral administration of single dose of aqueous extract of Qurse
nazla in Swiss albino mice shows that the formulation was well tolerated by the
animals and no abnormality was observed in the general behaviour of the
animals and no overnight mortality was recorded. Herbs and supplements can
be toxic when used for inappropriate indication, or prepared inappropriately, or
used in large excessive dosages or for a prolonged duration of time. Since it
is polyherbal formulation other ingredients present in formulation helps in
reducing the toxic effect of active component.
Sub-Acute Toxicity Study
The values of all the parameters including liver functions, renal functions
hematology and organ body weight ratio found to be normal as compared to
control group. The effects of the studied drug on organ body weight ratio in
control and treated animals are presented in Table 4. There were no significant
changes observed in organ body weight ratio of the control and the animals
treated with various doses. Table 1 and 2 is a summary of the results of the
effects of the drug on the biochemical parameters. There were no significant
changes in AST and ALT levels in all the treated animals compared with the
control. Similarly lipid profile of treated animal was found to be normal as
compared to control group. Table 3 reflects the values for hematological
parameters of treated group as compared to control showed no significant
changes in Hb, TLC, ESR, and % lymphocyte except % polymorph count in
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115Hippocratic Journal of Unani Medicine
Table 1: Effect of Aqueous Extract of Qurse- Nazla on Liver Function & Renal Function
Test in Albino Rats
Groups Liver Function Test Renal Function Test
SGOT SGPT SALP Serum Urea Serum
Unit/L Unit/L KAU Mg/dl Creatinine
Mg/dl
Group I
Normal Control 178.07±15.39 47.45±3.83 164.74±20.08 33.70 ± 4.03 1.29 ± 0.07
(n = 7)
Group II
Drug treated 177.09±14.52 45.40±2.43 166.85±19.30 33.56± 2.41 1.42 ± 0.06
1g /kg(n = 7) p = 0.9638 p = 0.694 p = 0.9409 p= 0.9767 p = 0.1839
Group III
Drug treated 176.02±13.50 47.86±4.85 169.42±17.42 34.83± 4.90 1.38±0.05
3g /kg(n = 7) p = 0.9219 p = 0.9482 p = 0.8632 p = 0.8616 p = 0.3161
Values are expressed as Mean±SEM
n = number of rats in a group.
P* < 0.05, P** < 0.01, P*** < 0.001
Table 2: Effect of Aqueous Extract of Qurse- Nazla on Lipid Profiles in Albino Rats
Groups Serum Cholesterol Serum HDL Serum Triglyceride
Mg/dl Mg/dl Mg/dl
Group I
Normal Control 47.12± 2.95 10.82±1 .30 43.60± 3.34
(n = 7)
Group II
Drug treated 48.76±4.01 11.32±0.68 41.20±4.06
1g /kg p = 0.7475 p = 1.00 p = 0.6562
(n = 7)
Group III
Drug treated 49.31±3.96 12.17±1.55 43.56±5.87
3g /kg p = 0.6653 p = 1.00 p = 0.9554
(n = 7)
Values are expressed as Mean±SEM
n = number of rats in a group.
P* < 0.05, P** < 0.01, P*** < 0.001
group III animals where p value = 0.0319. This is not quite significant to effect
safety of the drugs. It can be concluded on the basis of above observation that
drug is quite safe.
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116Hippocratic Journal of Unani Medicine
Table 3: Effect of the Aqueous Extract of Qurse- Nazla on Haematological Parameters
in Albino Rats
Groups Haemoglobin TLC / ESR DLC
gm % Cumm mm/hr
% Polymorph % Lymphocyte
count count
Group I
Normal Control 14.08±0.20 4264.28±147.88 1.71±0.28 30.85±3.23 65.14±2.46
(n = 7)
Group II
Drug treated 13.88±0.19 4350.00±186.56 1.57±0.29 36.57±2.91 58.00±3.70
1g kg p = 0.4824 p = 0.7520 p = 0.7344 p = 0.2129 p = 0.1340
(n = 7)
Group III
Drug treated 14.65±0.33 4157.14±4157.14 1.42±0.29 40.85±2.56 57.71±3.88
3g /kg p = 0.1654 p = 0.9719 p = 0.4857 p* = 0.0319 p = 0.1318
(n = 7)
Values are expressed as Mean±SEM
n = number of rats in a group.
P* < 0.05, P** < 0.01, P*** < 0.001
Table 4: Effect of the Aqueous Extract of Qurse-Nazla on Organ to the Body Weight
Ratio on Albino Rats
Groups Organ weight in g/100 g body weight
Liver Heart Kidney Spleen
Group I
Normal Control 4.50± 0.10 0.50± 0.01 0.99 ± 0.03 0.51± 0.07
(n = 7)
Group I
Drug treated 4.88±0.22 0.51± 0.01 1.15± 0.08 0.57 ± 0.03
1 g /kg p = 0.1418 p = 0.4930 p = 0.0857 p = 0.4461
(n = 7)
Group I
Drug treated 4.80± 0.23 0.50± 0.01 1.11 ± 0.05 0.52± 0.01
3g /kg p = 0.2547 p = 1.000 p = 0.0620 p = 0.889
(n = 7)
Values are expressed as Mean±SEM
n = number of rats in a group.
P* < 0.05, P** < 0.01, P*** < 0.001
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117Hippocratic Journal of Unani Medicine
Conclusion
The formulation was well tolerated by the animals and no abnormality was
observed in the general behavior of the animals and no overnight mortality
was recorded. There were no finding of any organ toxicity and hematological
changes as laboratory findings were normal. It can be concluded on the basis
of above observation that drug is quite safe in animals.
Acknowledgement
The authors would like to express their gratitude to Prof. Shakir Jamil, Director
General, Central Council for Research in Unani Medicine, New Delhi, for
providing research facilities to carry out this work.
References
Anonymous, 1993. Research Guidelines for Evaluating the Safety and Efficacy
of Herbal Medicine, Manila.WHO, pp. 35-40.
Bessay, O.A., Lowry, O.H., Brock, M.J., 1946. A method for the rapid
determination of alkaline phosphates with five cubic millimeters of serum.
J Biol.Chem. 164:321-329.
Bowers, L.D., Wong, E.T., 1980. Kinetic serum creatinine assay I. The role of
various factors in determining specificity. Clin.Chem. 26(5):551-554.
Burstain, M., Scholnic, H.R., Morphin, R., 1970. Rapid method for the isolation
of lipoprotein from human serum by precipitation with polyanions. J. Lipid.
Res 11:583-595.
Macgowan, M.W., Artiss, J.D., Strangberg, D.R., Zak, B., 1983. A peroxidase-
coupled method for the colorimetric determination of serum triglycerides.
Clin Chem. 29(3): 538-542.
Mukherjee, K.L., 1990. Medical Laboratory Technology, 3rd Edition, Tata
McGraw-Hill Publishing Company Limited, New Delhi, pp. 228-30.
Newcomer, H.S., 1919. Absorption Spectra of Acid Hematin, Oxyhemoglobin
and carbon monoxide hemoglobin: A new hemoglobinometer. J .Biol.chem.
37:465-496.
OECD, 2000. Guidance Document on the Recognition, Assessment and Use
of Clinical Signs as Humane Endpoints for Experimental Animals Used in
Safety Evaluation. Environmental Health and Safety Monograph Series on
Testing and Assesment No 19.
Plum, P., 1936. Accuracy of hematological counting method. Acta. Med.
Scandinav 90:342-364.
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118Hippocratic Journal of Unani Medicine
Reitman, S., and Frankel, S., 1957. A colorimetric method for determination of
serum glutamic oxaloacetic and glutamic pyruvic transaminases.
Am.J.Clin.Pathol. 28:56-63.
Roeschlau, P., Bernt, E., Gruber, W., 1974. Enzymatic determination of total
cholesterol in serum. Z Klin. Chem, Klin Biochem 12(5): 226.
Tiffany, T.O., Jansen, J., Burtis, C.A., Overton, J.B., Scott, C.D., 1972. Enzymatic
kinetic rate and end- point analyses of substrate, by use of a GeMSAEC
fast analyzer. Clin. Chem 18:829-840.
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119Hippocratic Journal of Unani Medicine
TAbstract
udri Surkh botanically equated to seeds of Cheiranthus cheiri
Linn. belongs to Cruciferae (Brassicaceae) family. In Unani System of Medicine
Tudri Surkh is used as stomachic, diuretic, expectorant, demulcent,
emmenagogue and also in the ailments of asthma, cough and fever. Seed oil
is applied locally for bruises, nervous and rheumatic pains. It is one of the
ingredients in the Unani formulations namely Majoon-e-Alkula, Majoon-e-
Regmahi, Khamira Gaozaban Ambari, Khamira Gaozaban Ambari Jadwar Ood
Saleeb Wala, Khamira Gaozaban Ambari Jawaharwala, Khamira Gaozaban
Sada. In view of its medicinal importance, the present study was conducted to
standardize the drug using pharmacognostic method, physico-chemical
parameters, TLC studies and WHO methods. Physico-chemical data observed
were moisture content (9.56%), total ash (6.60%), acid in-soluble ash (1.25%)
and solubility in alcohol (13.69 %) and water (20.01%). TLC studies of
chloroform and alcohol extracts showed various spots at 254nm, 366nm and
in visible light. The Quality control parameters such as microbial content (TBC,
TFC, Enterobacteriaceae, Salmonellae Spp. and Staphylococcus aureus) and
the heavy metals (Pb, Cd, As and Hg) were found within the permissible limits.
Aflatoxins (B1, B2, G1 and G2) and pesticide residues were not detected in the
drug Tudri Surkh.
Keywords: Tudri Surkh, Pharmacognostic, Physico-chemical, TLC, Quality
control parameters
Introduction
Plants plays a vital role in maintaining human health and improving the quality
of human life from thousands of years and serves to human the valuable
components of medicines, seasonings, beverages, cosmetics and dyes. Herbal
medicine contains natural substances that can promote health and reduce
illness. Nowadays researchers are focusing on plant research and it has
increased all over the world. Enormous evidence has been collected to show
immense potential of medicinal plants used in various traditional systems (Pratap
et al., 2013).
The seed of Cheiranthus cheiri Linn. is known as Tudri Surkh in Unani System
of Medicine. In Unani medicine the seeds are used as stomachic, diuretic,
expectorant, demulcent, emmenagogue and also used in the ailments of asthma,
cough and fever. Seed oil is applied locally for bruises, nervous and rheumatic
pains besides a tonic to improve the male reproductive system. Flowers are
Standardizationand QualityControlMethods ofUnani SingleDrug ‘TudriSurkh’(Cheiranthuscheiri Linn.)
1*D. Ramasamy,1Rampratap Meena,
1S. Mageswari,1P. Meera Devi Sri,
2Shamsul Arfin,1Syed Jameeluddin Ahmed
and2Syed Shakir Jamil
1Regional Research Institute
of Unani Medicine,
1, West Madha Church Street,
Royapuram, Chennai-600013
2Central Council for Research
in Unani Medicine,
61-65 Institutional Area,
Janakpuri, New Delhi-110058
July - September 2014, Vol. 9 No. 3, Pages 119-125
1*Author for correspondence
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120Hippocratic Journal of Unani Medicine
used in paralysis and impotence (Khare, 2007; Kritikar and Basu, 1998; Chopra
et al., 2006). Cheiranthus cheiri is a shrub like herb, indigenous in the North
Temperate zone, Central and Northern Europe; it is cultivated in Indian gardens
as an ornamental plant (wall flower).
Materials and Methods
Collection of drug
Seeds were collected from raw drug dealers, Chennai and identified by the
botanist and compared with the herbarium specimen of RRIUM, Chennai
(Specimen No. 00165).
Pharmacognostical studies
Botanical identification of the fruit was carried out using available literature
(Kritikar and Basu, 1998; Khare, 2007; Hooker, 1999). The pharmacognostical
studies such as macroscopical, microscopical and powder microscopy were
carried out using standard method (Johansen, 1940). Free hand sections of
the fruit were taken, microscopical drawings made using Camera Lucida and
observations recorded.
Physico-chemical parameters
Physico-chemical parameters like foreign matter, total ash, acid in-soluble ash,
loss on drying at 105°C, solubility in alcohol and water were carried out as per
standard method (Anonymous, 1998).
TLC analysis
Preparation of extract
The powder of the drug (2g) was extracted using 30ml of chloroform and
alcohol extracts were concentrated upto 10ml in a standard flask. These extracts
were used for the TLC studies.
The TLC profile of chloroform and ethanol extracts were performed using pre-
coated silica gel 60 F254 TLC plate (E. Merck) as adsorbent. TLC studies of
both extracts were carried out using solvent systems like toluene: ethyl acetate:
Acetic acid (8: 2: 0.2) and toluene: ethyl acetate (1: 1) respectively. After
drying, the plates were examined under UV – 254nm and 366nm and observed
the spots. Further the plates were dipped in vanillin-sulphuric acid reagent
followed by heating at 105ºC till appeared the bright spots appeared (Wagner
et al., 1984; Sethi, 1996).
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121Hippocratic Journal of Unani Medicine
Quality control parameters
The WHO parameters like microbial load, heavy metals, aflatoxin and pesticide
residues were carried out using standard methods of WHO & AOAC guidelines
(Anonymous, 1997, 1998, 2000).
Results and Discussion
Pharmacognostic studies
Macroscopic: Seeds are reddish brown, bright, 2.5 to 3.5mm long, 1.5 to 2 mm
wide, mucilaginous with warty surface; cotyledons incumbent, non-endospermic
with large embryo, musky odour and mucilaginous taste (Fig. 1 & 2).
Microscopic: T.S. of seed shows, epidermis consisting of single layer of
rectangular, flattened, thin walled cells containing colourless concentrically
striated mucilage; sclerenchyma cells palisade like consisting of single layer of
non-lignified cells with their radial and inner tangential walls thickened looks
like beaker shaped cells; pigmented cells consisting of single layer of elongated
parenchyma cells filled with yellowish brown contents; single layer of thick
walled cells followed by a layer of crushed parenchyma cells; cotyledons and
embryo consisting of oval to polygonal, thin-walled, parenchyma cells containing
aleurone grains and oil (Fig. 3,4 & 5).
Fig. 3: T S of Seed
A diagrammatic sketch
Fig. 1: Seeds Fig. 2: Seed - surface view
Abbreviation: EP - Epidermis; SCL - Sclerenchyma; CT - Cotyledons;
MUEP - Mucilaginous epidermis; PIGC - Pigmented cells; TWC - Thick walled cells
Fig. 4: T. S. of seed Fig. 5: T S of cotyledons
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122Hippocratic Journal of Unani Medicine
Powder Microscopy: Reddish brown, sclerenchyma cells in surface view,
epidermal cell is surface view with mucilage, thick walled cells in surface view;
elongated pigmented cells in surface view, cotyledonary parenchyma cells in
surface view
(Fig. 6).
Chemical analysis
Analytical data shows 9.56 % of moisture content. Ash content of the drug
was 6.59 % and 1.25 % of acid in-soluble ash shows the siliceous matter in
the plant. Alcohol soluble extractives represent the extraction of polar constituents
like phenols, tannins, glycosides, alkaloids and flavonoids. The water soluble
extractive denotes the presence of inorganic contents. The results of physico-
chemical parameters are shown in Table (1).
Table 1: Physico-chemical parameters of Tudri Surkh
S.No. Parameters Results (n=3)±S.D.
1. Foreign matter (%) Nil
2. Loss on drying at 105ºC (%) 9.56
3. Ash (%) 6.59
4. Acid insoluble ash (%) 1.25
5. Alcohol soluble extractives (% 13.69
6. Water soluble extractives (%) 15.81
Fig. 6: Powder microscopy
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123Hippocratic Journal of Unani Medicine
Thin Layer Chromatography
The Rf values of the TLC analysis of chloroform and alcohol extracts are
shown in Table - II and III. The plates were visualized using vannilin-sulphuric
acid reagent and heated at 105º till appear the colored spots. The TLC of the
chloroform extract at UV- 254 nm showed 5 spots, UV-366 nm showed 3 spots
and 5 spots showed after derivatization with vanillin - sulphuric acid (Fig. 7).
Alcohol extract showed at UV-254 nm 4 spots, UV-366 nm showed 3 spots and
after derivatization with vanillin – sulphuric acid showed 6 spots (Fig. 8).
Quality control parameters
The microbial load and heavy metals were found within the permissible limit
(Table – 4 and 5).
Fig. 7: Chloroform extract
Solvent system: Toluene : Ethyl acetate
: Acetic acid (8 : 2 : 0.2)
Detector: V. S. Reagent
Fig. 8: Alcohol extract
Solvent system: Toluene : Ethyl acetate
(1 : 1)
Detector: V. S. Reagent
Table 2: TLC data of the chloroform extract of Tudri Surkh
Solvent system Rf Values
UV 254 nm UV 366 nm V. S. Reagent
0.78 Pink 0.63 Blue 0.56 Dark grey
0.63 Light pink 0.26 Pale blue 0.45 Violet
0.49 Light pink 0.16 Blue 0.39 Grey
0.23 Pink 0.26 Violet
0.16 Pink 0.12 Grey
Toluene : Ethyl
acetate : Acetic acid
(8 : 2 : 0.2)
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124Hippocratic Journal of Unani Medicine
Table 3: TLC data of the alcohol extract of Tudri Surkh
Solvent system Rf Values
UV 254nm UV 366nm V. S. Reagent
0.91 Light pink 0.72 Blue 0.91 Grey
0.72 Pink 0.39 Pale blue 0.72 Violet
0.39 Pink 0.14 Blue 0.56 Light grey
0.14 Pink 0.39 Light grey
0.34 Violet
0.14 Violet
Toluene: Ethyl acetate
(1 : 1)
Table 4: Microbial load
S.No. Parameter Analyzed Results WHO Limits
1 Total Bacterial Count 2 x 10 2 CFU / gm 105 CFU / gm
2 Total Fungal Count Ansent 103 CFU / gm
3 Enterobacteriaceae Absent 103 CFU / gm
4 Salmonella Spp. Absent Nil
5 Staphylococcus aureus Absent Nil
Table 5: Heavy metals
S.No. Parameter Analyzed Results WHO & FDA Limits
1 Arsenic Nil 10 ppm
2 Cadmium Nil 0.3 ppm
3 Lead 0.0031 10 ppm
4 Mercury Nil 1.0 ppm
The aflatoxin such as B1, B2, G1 & G2 and analysed pesticide residues such as
organo chlorine group, organo phosphorus group, acephate, chlordane,
dimethoate, endosulphan, ethion, endosufon sulphate, fenthion, lindane,
methoxychlor, phorate sulfoxide and phorate sulfone were not detected from
the drug.
Conclusion
The evaluated standards such as macroscopic, microscopic, physico-chemical,
TLC analysis and quality control parameters were derived and described are
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125Hippocratic Journal of Unani Medicine
of diagnostic impor-tance in authentication and quality control of the seeds of
Cheiranthus cheiri.
Acknowledgement
The authors are deeply indebted to the Director General, CCRUM, New Delhi,
for providing necessary research facilities and encouragement for this study.
References
Anonymous, 1997. Official Analytical Methods of the American Spice Trade
Association (ASTA). Inc. 4th edn., New Jersey, pp. 149-152.
Anonymous, 1998. Quality Control Methods for Medicinal Plant Materials. World
Health Organisation, Geneva, pp.10-31, 61-63.
Anonymous, 2005. Official Methods of Analysis of AOAC International. In: Horwitz
W, Latimer, G.W. (eds). 18th Edn. AOAC International: Maryland, chapter 10
pp.18-23 and chapter 3, pp. 10-11.
Anonymous, 2006. National Formulary of Unani Medicine, Part – I. Ministry of
Health & Family Welfare, Department of AYUSH, Govt. of India, New Delhi.
Pratap, Bhanu, G.S. Chakraborthy, Nandini, Mogha, 2013. Complete aspects
of Alstonia scholaris. International Journal of Pharm.Tech. Research 5(1):
17-26.
Hooker, J.D., 1999. The Flora of British India, Vol. I. Bishen Singh Mahendra
pal Singh, Dehra Dun, p. 132.
Johansen, D.A., 1940. Plant Microtechnique Mc. Graw Hill Book Company Inc.,
New York and London, pp. 181 - 186.
Khare, C.P. (Ed.), 2007. Indian Medicinal Plants, An Illustrated Dictionary,
Springer International Edition, pp. 140–141.
Kritikar, K.R. and Basu, B.D., 1998. Indian Medicinal Plants, Vol. 1. Bishen
Singh Mahendra Pal Singh, Dehra Dun, IInd Edition, pp.143–145.
Chopra, R.N., Nayar, S.L. and Chopra, I.C., 2006. Glossary of Indian Medicinal
Plants, National Institute of Science Communication and Information
Resources. Council of Scientific & Industrial Research, New Delhi, India,
pp. 60–61.
Sethi, P.D., 1996. High Performance Thin Layer Chromatography. CBS Publisher
and Distributors, New Delhi.
Wagner, H., Bladt, S.A., 1996. Thin Layer Chromatography Atlas. In: Plant
Drug Analysis. 2nd edn. Springer-Verlag Berlin Heidelberg, Germany.
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126Hippocratic Journal of Unani Medicine
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127Hippocratic Journal of Unani Medicine
TAbstract
he Unani system of medicine prescribes large number of
classical herbal formulations to cure the different types of diseases. Jawarish-
e-Kafoor Qawi a Unani herbal formulation is prepared in combination of
ingredients like Kafoor, Zafran, Jauzbuwa, Filfil Siyah, Zanjabeel, Bisbasa,
Darchini, Narmushk, Qirfa, Filfilmoya, Faranjmushk and Qand Safaid. The Unani
Physicians prescribes the drug Jawarish-e-Kafoor Qawi to cure the ailments of
Zof-e-Meda (Weakness of the stomach) and Nafkh-e-Shikam (Flatulence in the
stomach). At present no pharmacopoeial standards on drug is available and
it is basic requirement for the research on quality control of this drug. There
is lack of standardization and proper documentation of Unani drugs. Based on
the available sources an attempt is made to evaluate the drug on
pharmacopoeial parameters to develop standards for the drug Jawarish-e-
Kafoor Qawi. To evaluate the pharmacopoeial parameters of the drug, various
parameters like powder microscopy, moisture content, ash values, bulk density,
pH values, extractive values, TLC/HPTLC finger printing and other quality
control parameters viz. heavy metals, microbial content, aflatoxins and pesticide
residues are performed. The evaluated data will help to lay down
pharmacopoeial standards for the drug ‘Jawarish-e-Kafoor Qawi’.
Keywords: Jawarish-e-Kafoor Qawi, Powder microscopy, Physico-chemical,
TLC/HPTLC, WHO parameters
Introduction
Jawarish-e-Kafoor Qawi (Anonymous, 2006) is one of the ancient commonly
used classical Unani formulations. This poly herbal formulation consists of 12
ingredients (Table 1). This drug is prescribed for the treatment of Zof-e-Meda
(weakness of the stomach) and Nafkh-e-Shikam (flatulence in the stomach)
disorders. The development of traditional medicines particularly Unani medicines
with the perspective of safety, efficacy and quality will not only to preserve the
traditional heritage but also to rationalize the uses of Unani medicines in the
health care.
Standardisation of Unani herbal formulations is necessary step to assess the
quality of drugs. Due to lack of Standard Operating Procedures (SOP’s) and
quality control methods, there are batch to batch variations among the similar
formulations. Pharmacopoeial study of a drug is an essential requirement to
establish the presence of each ingredient in the formulations (Bandaranayake,
Development ofQualityStandards onJawarish-e-Kafoor Qawi – AClassical UnaniFormulation#
1*Rampratap Meena,2S. Mageswari,2D. Ramasamy,
2P. Meera Devi Sri,3Shamsul Arfin,
3Aminuddin
and4Nitin Rai
1Drug Standardisation Research
Institute,
PLIM Campus, Sanjay Nagar,
Ghaziabad-201002
2Regional Research Institute
of Unani Medicine,
1, West Madha Church Street,
Royapuram, Chennai-600013
3Central Council for Research
in Unani Medicine,
61-65 Institutional Area,
Janakpuri, New Delhi-110058
4Pharmacopoeial Laboratory
for Indian Medicine,
Kamla Nehru Nagar,
Ghaziabad-201002
July - September 2014, Vol. 9 No. 3, Pages 127-140
# Accepted for presentation in ‘National Seminar on Opportunities in Medicinal Plant Research’,
Jadavpur University, Kolkata-32; November 29-30, 20141*Author for correspondence
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128Hippocratic Journal of Unani Medicine
2006; Myers and Cheras, 2004). The present study is an attempt to evaluate
the pharmacopoeial studies of the drug by applying modern parameters such
as microscopical, physico-chemical, thin layer chromatography and WHO
parameters such as microbial load, aflatoxin, heavy metals and pesticide residue.
Material and Methods
To evaluate the pharmacopoeial studies of Jawarish-e-Kafoor Qawi a systematic
scheme of standardization was developed.
(i) Ingredients authentications
Genuine raw drugs namely Kafoor, Zafran, Jauzbuwa, Filfil Siyah, Zanjabeel,
Bisbasa, Darchini, Narmushk, Qirfa, Filfilmoya, Faranjmushk and Qand Safaid
of the formulation were procured from raw drugs dealers of Chennai and Delhi
(Fig. 1). The raw drugs were authenticated as per pharmacopoeial and other
official standards (Anonymous, 2004, 2007, 2008 & 2009).
(ii) Drug formulation
The ployherbal semisolid drug was prepared in different batches at Laboratory
scale as per the ingredients composition and guidelines of NFUM, Part – IV
(Anonymous, 2006) (Table 1).
(iii) Powder microscopy
The drug sample (5g) was weighed and mixed with 50ml of water in a beaker
with gentle warming, till the sample completely dispersed in water. The mixture
was centrifuged and decanted the supernatant. The sediment was washed
several times with distilled water, centrifuged again and decanted the
supernatant. A few mg of the sediment was taken and mounted in glycerine.
A few mg was taken in watch glass and added few drops of phloroglucinol and
concentrated hydrochloric acid, mounted in glycerine. The microscopic salient
features of the drug were observed in different mounts (Wallis, 1997; Johansen,
1940).
(iv) Physico-chemical analysis
The physico-chemical methods viz., moisture content, ash values, solubility in
different solvents, pH values, bulk density and sugar content etc., are useful
tools in standardisation of a herbal product for maintaining batch to batch
consistency. The drug samples were subjected for the standardisation of
physico-chemical parameters and analysed as per the standards method
(Anonymous, 1987).
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129Hippocratic Journal of Unani Medicine
Fig. 1: Ingredients of Jawarish-e-Kafoor Qawi
Filfil Siyah Zanjabeel BisbasaPiper nigrum Linn. Zingiber officinale Rosc. Myristica fragrans Houtt.
Fruit Rhizome Arillus
Kafoor Zafran JauzbuwaCinnamomum camphora (L.) Crocus sativus Linn. Myristica fragrans Houtt.
Nees & Eberm Dried stigmas and top of styles EndospermNatural Camphor
Filfilmoya Faranjmushk SugarPiper longum L. Ocimum sanctum L.
Fruit Seed
Darchini Narmushk QirfaCinnamomum zeylanicum Blume. Mesua ferrea Linn. Cinnamomum cassia Blume.
Inner stem bark Stamens Stem bark
(v) TLC/HPTLC finger printing
The formulations of the three batch samples were extracted with chloroform
and alcohol. The extracts were concentrated and made up to 10 ml in a
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130Hippocratic Journal of Unani Medicine
volumetric flask separately. These solutions were used for the TLC/HPTLC
finger print analysis.
The TLC/HPTLC finger print analysis of chloroform and alcohol extracts of the
formulations were performed using aluminium plate precoated with silica gel 60
F254 (E.merck) employing CAMAG Automatic TLC sample - IV applicator. The
chromatogram were developed using the developing systems toluene: ethyl
acetate (9: 1) and toluene: ethyl acetate (6: 4) for chloroform and alcohol
extracts respectively. The plates were dried at room temperature to record the
image of the plates at UV-254 nm, UV-366 nm using TLC visualizer and the
plates were scanned at 254 nm to record the finger print spectrum using TLC
Scanner - IV. Finally the plate were dipped in vanillin-sulphuric acid and heated
at 105° till coloured spots appeared (Wagner, and Bladt, 1984; Sethi, 1996).
(vi) Other quality control parameter
The usage of herbal products along with higher safety margins, WHO has
taken necessary step to ensure quality control parameters with the modern
techniques and application of suitable standards. The microbial load and heavy
metal parameters were carried out as per the WHO guidelines (Anonymous,
1998). Aflatoxin and pesticide residues were carried out by standard methods
(Anonymous, 2000).
Obseravtions
Jawarish-e-Kafoor Qawi is a dark brown semi-solid product with sweetish bitter
taste.
(i) Pharmacognostical observation (Powder microscopy):
The diagnostic characteristics of cellular elements in respect of each ingredients
is in Table 1 and Fig. 2.
(ii) Chemical analysis
The physico-chemical data such as moisture content was obtained in the drug
19.57%. The alcohol soluble extractive (44.56%) might be due to the extraction
of polar chemicals constituents and the water soluble extractives (65.54%)
indicate the presence of inorganic constituents. The obtained data are shown
in Table 2.
(iii) Thin Layer Chromatography analysis
The chloroform and alcohol extract of all the three batch samples showed
identical spots in UV – 254nm and 366nm ranges and the Rf values of both
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131Hippocratic Journal of Unani Medicine
Table 1: List of the raw drugs and cellular elements of Jawarish-e-Kafoor Qawi
Formulation
S. Unani name Unani Name Part used Quantity Salient features of the drug
No.
1. Kafoor Cinnamomum Natural 25 g. –
API-VI camphora (L.) camphor
Nees & Eberm.
2. Zafran Crocus Stamens & 25 g. Pollen grains size upto 120μ, spherical
UPI-VI sativus Linn. Stigmas in outline with clear exine and intine
3. Jauzbuwa Myristica Endosperm 25 g. Endosperm cells filled with numerous
UPI-I fragrans starch grains, crystalline fat and large
Houtt. aleurone grains (crystalloid proteins
upto 40μ), perisperm cells filled with
reddish brown contents
4. Filfil Siyah Piper nigrum Fruit 25 g. Stone cells polygonal interspersed
UPI-IV L. among parenchyma cells with circular
lumen, perisperm cells isolated or in
groups with angular walls filled with
aleurone grains and minute calcium
oxalate crystals
5. Zanjabeel Zingiber Rhizome 25 g. Isolated starch grains, simple oval to
UPI-I officinale round shaped measuring upto 70μ,
Rosc hilum eccentric, lamellae distinct;
non-lignified septate fibres upto 50μ,
reticulate vessels and fragments of
reticulate vessels upto 70μ;
parenchyma cells filled with abundant
starch grains
6. Bisbasa Myristica Arillus 25 g. Thick walled elongated parenchyma
UPI-VI fragrans cells in surface view upto 50μ wide
Houtt.
7. Darchini Cinnamomum Inner stem 25 g. Fibres thick walled lignified with
UPI-I zeylanicum bark striated walls and narrow lumen of
Blume. length upto 1000μ and breadth upto
30μ, stone cells with horse shoe
shaped thickenings upto 70μ
8. Narmushk Mesua Stamens 25 g. Tricolporate golden yellow pollen
UPI-IV ferrea Linn grains upto 50μ
9. Qirfa UPI-III Cinnamomum Stem bark 25 g. Fibres thick walled lignified with
cassia Blume. striated walls and narrow lumen of
length upto 1000μ and breadth upto
40μ and very large stone cells upto
200μ, stone cells with horse shoe
shaped thickenings upto 70μ
10. Filfilmoya Piper longum Fruit 25 g. Parenchyma cells with elongated
API-IV L. spindle shaped stone cells; perisperm
cells isolated or in groups with angular
walls filled with aleurone grains and
minute calcium oxalate crystals
11. Faranjmushk Ocimum Seed 25 g. Fragments of irregular shaped thick
API-IV sanctum L. walled epidermal cells
12. Qand Safaid Sugar – 800 g. –
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132Hippocratic Journal of Unani Medicine
Fig. 2: Powder microscopy of Jawarish-e-Kafoor Qawi
Jauzbuwa Bisbasa
Elongated thick walledparenchyma cellsZafran
Pollen grains
Endosperm cells filledwith starch grains Perisperm cells
Filfil Siyah
Parenchyma cellswith stone cells
Filfilmoya
Parenchyma cells withspindle shaped stone cells
Filfil Siyah & Filfilmoya
Perisperm cells with angularwalls filled with starch grains
Narmushk ZanjabeelPollen grains Reticulate vessels Septate fibres Starch grains
Parenchyma cellsfilled with starch grains
Qirfa / Darchini
FaranjmushkFibres Stone cells
Thick walled irregularepidermal cells
the extracts are shown in Table 3 and 4. The plates were dipped in vanillin-
sulphuric acid and heated at 105ºC till appeared coloured spots.
(iv) Quality control parameters
The evaluated quality control parameters such as microbial load and heavy
metals were found within the permissible limit in the drug shown in Table 5
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133Hippocratic Journal of Unani Medicine
Table 2: Physico-chemical parameters
Parameters Batch Number (n=3)
I II III
Extractives
Alcohol soluble matter 44.71% 44.17% 44.80%
Water soluble matter 65.31% 65.79% 65.52%
Ash
Total ash 0.89% 0.73% 0.62%
Acid insoluble ash 0.18% 0.28% 0.35%
pH values
1% Aqueous solution 5.36 5.48 5.39
10% Aqueous solution 4.19 4.35 4.41
Sugar estimation
Reducing sugar 33.61% 33.26% 33.72%
Non-reducing sugar 9.47% 9.09% 9.50%
Moisture 19.24% 19.83% 19.46%
Bulk Density 1.3203 1.3114 1.3152
Table-3: Rf values of the chloroform extract
Rf Values
Solvent UV-254 nm UV-366 nm After derivatisation with
System vanillin – sulphuric acid
reagent
0.95 Green 0.92 Fluorescent blue 0.93 Grey
0.80 Green 0.81 Blue 0.83 Violet
0.72 Green 0.78 Red 0.73 Violet
0.67 Green 0.69 Red 0.66 Grey
0.60 Green 0.64 Fluorescent blue 0.59 Grey
0.48 Green 0.60 Red 0.57 Grey
0.35 Green 0.56 Red 0.48 Pink
0.27 Green 0.50 Blue 0.37 Grey
0.21 Green 0.44 Red 0.32 Violet
0.16 Green 0.23 Green 0.27 Pink
0.14 Blue 0.15 Grey
Tolu
ene:
Eth
yl a
ceta
te (
9:1)
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134Hippocratic Journal of Unani Medicine
and 6. The other parameters like aflatoxins B1, B2, G1 and G2 and pesticide
residues - organo chlorine group, organo phosphorus group, acephate,
chlordane, dimethoate, endosulphan, endosulfan, endosulfon, ethion,
endosufon sulphate, fenthion, heptachlor, lindane, methoxychlor, phorate
sulfoxide and phorate sulfone were not detected from the drug samples shown
in Table 7 and 8.
Table 4: Rf values of the alcohol extract
Rf Values
Solvent UV-254 nm UV-366 nm After derivatisation with
System vanillin – sulphuric acid
reagent
0.92 Green 0.92 Red 0.92 Violet
0.88 Green 0.86 Pink 0.87 Brown
0.79 Green 0.78 Blue 0.78 Grey
0.72 Green 0.59 Green 0.69 Light grey
0.48 Green 0.51 Green 0.65 Violet
0.39 Green 0.47 Blue 0.58 Light grey
0.23 Light blue 0.49 Light grey
0.10 Pink 0.10 Grey
Tolu
ene:
Eth
yl a
ceta
te (
6:4)
Table 5: Analysis of Microbial load
S.No. Parameter Analyzed Results WHO Limits
1 Total Bacterial Count 300 CFU / gm 105 CFU / gm
2 Total Fungal Count < 10 CFU/ gm 103 CFU / gm
3 Enterobacteriaceae Absent / gm 103 CFU / gm
4 Salmonella Absent / gm Nil
5 Staphylococcus aureus Absent / gm Nil
Table 6: Estimation of Heavy Metals
S.No. Parameter Analyzed Results WHO & FDA Limits
1 Arsenic Not detected 10 ppm
2 Cadmium Not detected 0.3 ppm
3 Lead 0.0018 ppm 10 ppm
4 Mercury Not detected 1.0 ppm
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135Hippocratic Journal of Unani Medicine
TLC/ HPTLC finger print studies
(i) TLC/ HPTLC finger print studies of chloroform extract
The TLC studies of chloroform extract are tabulated in Table 3. All the three
batch samples shows identical spots in UV-254 nm, UV-366 nm and visible light
(after derivatised with vanillin – sulphuric acid reagent). In UV – 254, 366 nm
and visible light it shows 10, 11 and 11 spots respectively with different Rf
values (Fig. 3). The finger print of the chloroform extract shows 13 peaks of
Table 7: Estimation of Aflatoxins
S.No. Aflatoxins Results WHO Limits
1 B1 ND 0.05ppb
2 B2 ND 0.05ppb
3 G1 ND 0.05ppb
4 G2 ND 0.05ppb
ND = Not Detected
Table 8: Analysis of Pesticide Residues
S.No. Pesticide Residues Results Limits
1 Organo Chlorine group ND (DL 0.005mg/Kg)
2 Organo Phosphorus group ND (DL 0.005mg/Kg)
3 Acephate ND (DL 0.005mg/Kg)
4 Chlordane ND (DL 0.005mg/Kg)
5 Dimethoate ND (DL 0.005mg/Kg)
6 Endosulphan ND (DL 0.005mg/Kg)
7 Endosulfan ND (DL 0.005mg/Kg)
8 Endosulfon ND (DL 0.005mg/Kg)
9 Ethion ND (DL 0.005mg/Kg)
10 Endosufon sulphate ND (DL 0.005mg/Kg)
11 Fenthion ND (DL 0.005mg/Kg)
12 Heptachlor ND (DL 0.005mg/Kg)
13 Lindane ND (DL 0.005mg/Kg)
14 Methoxychlor ND (DL 0.005mg/Kg)
15 Phorate sulfoxide ND (DL 0.005mg/Kg)
16 Phorate sulfone ND (DL 0.005mg/Kg)
ND – Not detected
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136Hippocratic Journal of Unani Medicine
which peaks at Rf 0.17, 0.30, 0.38, 0.57, 0.62, 0.75, 0.82 and 0.91 were the
major peak whereas peaks at Rf 0.04, 0.07, 0.22, 0.44 and 0.67 were
moderately smaller peaks (Fig.4). The HPTLC densitometry chromatogram of
chloroform extract of three batch samples were recorded at 254 nm (Fig. 5).
UV- 254 nm UV- 366 nm Visible Light(After derivatisation with
vanillin–sulphuric acid reagent)
Fig. 3: TLC photos of chloroform extracts of three batch samples at different wavelength
of light
Fig. 4: HPTLC finger print profile for chloroform extract at 254 nm
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137Hippocratic Journal of Unani Medicine
Fig. 5: HPTLC densitometry chromatogram of chloroform extracts of three batch
samples at 254 nm
(ii) TLC/ HPTLC finger print studies of alcohol extract
The TLC studies of alcohol extract are tabulated in Table 4. All the three batch
samples shows identical spot in UV-254 nm, UV-366 nm and visible light (after
derivatised with vanillin – sulphuric acid reagent). In UV – 254, 366 nm and
visible light it shows 6, 8 and 8 spots respectively with different Rf values (Fig.
6). The finger print of the alcohol extract shows 13 peaks of which peaks at
Rf 0.54, 0.74, 0.81 and 0.90 were the major peak whereas peaks at Rf 0.08,
0.12, 0.25, 0.32, 0.37, 0.44, 0.60, 0.67 and 0.99 were moderately smaller
peaks (Fig. 7). The HPTLC densitometry chromatogram of alcohol extract of
three batch samples were recorded at 254 nm (Fig. 8).
Results and Discussion
The evaluated data in respect of powder microscopy, physico-chemical, TLC/
HPTLC fingerprint and other quality parameters provides analytical parameters
on classical Unani formulation Jawarish-e-Kafoor Qawi can be used for in
house quality control of drug as well for development of pharmacopoeial
standards on drug.
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138Hippocratic Journal of Unani Medicine
UV- 254 nm UV- 366 nm Visible Light
(After derivatisation with
vanillin – sulphuric acid reagent)
Fig. 6: TLC photos of alcohol extracts of three batch samples at different wavelength of
light
Fig. 7: HPTLC finger print profile for alcohol extract at 254 nm
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139Hippocratic Journal of Unani Medicine
Fig. 8: HPTLC densitometry chromatogram of alcohol extracts of three batch samples
at 254 nm
Acknowledgement
The authors are extremely thankful to Director General, CCRUM, New Delhi
and the Director, Pharmacopoeial Laboratory of Indian Medicine (PCIM/PLIM),
Ghaziabad, for their valuable guidance, encouragement and providing necessary
research facilities to carry out the present studies.
References
Anonymous, 1987. Physico-chemical standards of Unani Formulations Part –
II. CCRUM, Min. of Health & Family Welfare, New Delhi, pp. 300 - 317.
Anonymous, 1998. Quality Control Methods for Medicinal Plant Materials. World
Health Organization, Geneva, pp. 25 - 28.
Anonymous, 2000. Association of Official Analytical Chemists (AOAC), 17th
Edition.
Anonymous, 2004. The Ayurvedic Pharmacopoeia of India, Part-I, Vol.-IV (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp.
105-106; 146-147.
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140Hippocratic Journal of Unani Medicine
Anonymous, 2006. National Formulary of Unani Medicine, Part – IV (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, p. 59.
Anonymous, 2007. The Unani Pharmacopoeia of India, Part-I, Vol.-I (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp. 26-
27; 38-39; 88-89.
Anonymous, 2007. The Unani Pharmacopoeia of India, Part-I, Vol.-III (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp. 82-
83.
Anonymous, 2007. The Unani Pharmacopoeia of India, Part-I, Vol.-IV (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp. 38-
39; 98-99.
Anonymous, 2008. The Ayurvedic Pharmacopoeia of India, Part-I, Vol.-VI (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp.
210-211.
Anonymous, 2009. The Unani Pharmacopoeia of India, Part-I, Vol.- VI (English
Edition). Govt. of India, Min. of Health & Family Welfare, New Delhi, pp. 23-
24; 101-102.
Bandaranayake WM, 2006. Quality control, screening, toxicity and regulation
of herbal drugs. Modern Phytomedicines, pp. 25 - 57.
Johansen, D.A., 1940. Plant Microtechnique. Mc. Graw Hill Book Company Inc.,
New York and London, pp. 181-186.
Myers, S.P., Cheras, P.A., 2004. The other side of the coin: safety of
complementary and alternative medicine. Medical J. Australia 181: 222 -
225.
Sethi, P.D., 1996. High Performance Thin Layer Chromatography, Vol. X. (1st
Edition). CBS Publishers and Distributors, New Delhi, pp. 1-56.
Wagner, H., Bladt, S., and Zgainski, E.M., 1984. Plant Drug Analysis: A Thin
Layer Chromatography Atlas (2nd Edition). Springer-Verlag, Germany.
Wallis, R.E., 1997. Text Book of Pharmacognosy, 5th Edition. CBS Publishers
& Distributors, Delhi, pp. 494 – 496.
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141Hippocratic Journal of Unani Medicine
Short Research Communication:
Overview of Catharanthus roseus (L.)G. Don Alkaloids
Catharanthus roseus (L.) G.Don; syn. Vinca rosea L.
Family : Apocynaceae
English name : Periwinkle
Hindi name :Sadabahar, Baramasi
Introduction
A tropical perennial herb, hardy and resistant to attack by fungi or insects. It
grows wild but now cultivated scientifically for ornamental and medicinal use.
Flowers white or deep rose coloured; follicles long. Earlier civilisations knew
about its use in medicine.
The plant originated from Malagasy (Madagascar), then spread to Mozambique,
followed by several South East Asian countries viz., South Vietnam, Philippines
and Australia, including India and Ceylon (Tiwari et al., 2011).
Alkalodial Composition
Catharanthus roseus (L.) G.Don contains two important classes of compounds,
viz; alkaloids and tannins. Although it is a poisonous plant, but under close
medical supervision its alkaloids have been employed successfully for the
treatment of cancers viz; Hodgkin’s carcinoma and leukemia.
There are over 130 organic constituents present in Catharanthus roseus (L.)
G.Don plant. These have indole or di-hydro-indole moieties present in their
molecular structure, which exhibited oncological activity. The chief alkaloids
which we shall focus upon here are known as Vinblastine and Vincristine. They
occur together naturally and the yield of the former alkaloid is pre-dominating.
The 17-de-acetyl vinblastine has been synthetically converted to the
corresponding lactone, which was found to be associated with chemotherapeutic
activity.
Applications
The alkaloidal components of Catharanthus roseus (L.) G.Don have been
claimed to be good for brain health. These alkaloids help in improving the
supply of nutrient (glucose) and oxygen to the brain, which it can effectively
utilize. Additionally these components are highly effective in the abnormal
July - September 2014, Vol. 9 No. 3, Pages 141-145
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142Hippocratic Journal of Unani Medicine
coagulation of blood and in raising the level of serotonin. This molecule is well
known blood neurotransmitter in the Central Nervous System (CNS); its vital
function is to regulate the heart function, memory including sleep and appetite.
Lack of serotonin may lead to several mental disorders like schizophrenia. The
other application of Catharanthus roseus (L.) G.Don alkaloids are;
• Hypotensive
• Sedative
• Tranquiliser
• Anticancer
• Relief from pain in muscles and wasp stings
• Provides relief to depression of the CNS
• Stop bleeding from nose/gums
• Relief to mouth ulcers and sore throat
• Decrease glucose concentration in blood (hypoglycaemia) (specially
from plant leaves)
• Controls diarrhea
• Taken internally it produces memory enhancing action
• Reduces the inflammation of urinary bladder, stomach and intestines
• Wound healing properties
According to literature reports there are over 100 alkaloids isolated from the
various parts of the plant (viz; leaves, stem, root, root bark etc.).Amongst them
two medicinally important alkaloids may be mentioned here viz; vinblastine and
vincristine. The yield of vincristine is extremely small (0.00025%) in comparison
to vinblastine (root : 0.78-1.22; root bark : 4.5-9.00)% respectively (Tiwari et
al., 2001). Amongst the other bio-active alkaloids may be mentioned reserpine
,ajmalicine (raubasine), serpentine and vincamine. The laboratory synthesized
derivative of vincamine (alternatively known as vincopectine) is a well known
triple action medicine, viz; vaso-dilator, blood thinner and memory enhancer.
Praubasine has been reported to be useful in checking fragility of capillaries
(tiny thin walled blood vessels).
One outstanding property of Catharanthus roseus (L.) G.Don extract is that it
exhibits anticancer activity against numerous cell type in general and against
multi drug resistant cancer tumors in particular (Favretto et al., 2001). This
observation points to the fact that some components of the Catharanthus
roseus (L.) G.Don exerts a synergic effect which counters the multi drug
resistance offered by the tumors cells.
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143Hippocratic Journal of Unani Medicine
In this article we shall highlight the pharmaceutical uses of the two clinically
important alkaloids viz; vinblastine and vincristine used for the treatments of
cancers.
Vinblastine
This alkaloid exhibits quite a different and wide spectrum of activity, and is
used for the following types of cancers;
Hodkgin’s lymphoma, cancer of breast, head, lungs, neck, ovaries and testicles.
The vinblastine is biosynthesized in the plant by joining together two alkaloidal
residues(viz; Cathaanthine and vindoline).
According to the regimen of chemothraphy directed from the Hodgkin’s
lymphoma, vinblastine is incorporated into belomycin and metotrenote. This
allows the use of lower dose of belomycin thereby reducing the overall toxicity
and providing larger resting periods between chemotherapy cycles (Gobbi et
al., 2003).
Caution should however, be exercised in the use of vinblastine, since it is
associated with wide adverse side effects, namely, hair loss, stomach and
intestinal disorders, elevation of blood pressure, depression, involuntary and
painful contraction of muscles, giddiness, headache etc.
Vincristine
Vincristine is the second vinca alkaloid, which is used for the treatment of
cancer. This cytotoxic drug acts by inhibiting the ability of the cell to divide,
which in turn stops the proliferation of the disease .The alkaloid has quite a
different and wide spectrum of activity. It is employed for the acute lymphobic
cancers in children, malignant tumor (Favretto et al., 2001) and breast cancer.
The use of the extract of Catharanthus roseus (L.) G.Don on a rabbit led Beer
& Noble (Western Ontario, Canada) to a chance discovery. They noticed that
there was reduction of white blood cells (WBCs)and which decreased further
with the administration of higher concentration of the drug. Besides, the blood
platelets count, including the immunity of the animal fell considerably. There
was practically no resistance in the body of the rabbit and the animal died due
to infection of Pseudomonas sp. This led the investigators to conclude that
Catharanthus roseus (L.) G.Don. alkaloid could be used to cure leukaemia –
a medical condition wherein there is a large increase of WBCs than the normal.
This conclusion was found to be correct and was based on the positive results
obtained with the leukemic patients. The property of Croesus’s alkaloid to
suppress immunity could be exploited in organ transplant operations to overcome
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144Hippocratic Journal of Unani Medicine
the problem of ‘rejection’ by the body .At this stage a need was felt to devise
a method of obtaining vincristine alkaloid in sizeable amounts. Recourse was
taken to extractive procedure from vinca rosea, involving the use of tons
quantity of dried leaves. This, however, is a cumbersome and time consuming
process. Thanks to the efforts of synthetic organic chemists, who were able to
prepare the valuable vincristine alkaloid on a pilot plant scale, in order to cope
up with the demand of its use as a drug in cancer therapy as well as an
immuno-suppressant drug.
These two alkaloids (vinblastine and vincristine) are now available in the US
markets in the form of their respective sulphates.
Isolation of the alkaloids
The modern technique has been employed for the isolation of the above
mentioned alkaloid in a pure form;
Methods involving the;
• Selective or differential extraction.
• Method based on pH gradient based separation.
Flavonidal/terpenoidal composition
The flowers of Catharanthus roseus (L.) G.Don contained flavonids. The major
anthocyanidins present consist of Petunidin, malvidin, including kaempferol
and Quercetin (both are strong antioxidants). The whole plant is reported to
contain tri-terpenoid,viz ; ursolic acid, accompanied with loganin, deoxyloganin
severoside etc.
Acknowledgement
The authors thank Prof. G.S. Sandhu, Director, HIET, Ghaziabad, for the kind
interest shown by him during the present study.
References
Favretto, D., Piovan, A., Filippini, R., Caniato, R., 2001. Monitoring the production
yields of vincristine and vinblastine in Catharanthus roseus from somatic
embryogenesis. Semi-quantitative determination by flow- injection
electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom.
15(5): 364-9.
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145Hippocratic Journal of Unani Medicine
Gobbi, P.G., Brogila, C., Merli, F., Dell’Olio, M., Stelitanto, C., Lannito, E.,
Federico, M., Berte, R., Luisi, D., Molica, S., Cavalli, C., Dezza, L., Ascari,
E., 2003. Vinblastine, Belomycin and Methotrexate Chemotherapy plus
irradiation for Patients with Early Stage, Favorable Hodgkin Lymphoma.
Cancer 98 (11): 2393–2401.
Tiwari, D.N., Kumar, K. and Tripathi, A., 2001. SADABAHAR. Ocean book Pvt.
Ltd., Allahabad, p. 80.
July 2, 2014 Durga Nath Dhar
44 Duplex, Block-A
Rajat Vihar, Sector 62
NOIDA-201301
and
Shalin Kumar
Hi-Tech Institute of
Engg.& Technology
NH-24, Adhyatmik Nagar
Ghaziabad – 201015
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146Hippocratic Journal of Unani Medicine
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147Hippocratic Journal of Unani Medicine
HIPPOCRATIC JOURNAL OF UNANI MEDICINE
Instructions to contributors
1. The paper(s) should be submitted in duplicate. Submission of a paper will
be taken to imply that it is unpublished and is not being considered for
publication elsewhere.
2. Papers should be written in English language and typed with double
spacing on one side of A-4 size paper leaving top and left hand margin
at least 1" (One inch) wide. Length of the paper should normally not
exceed 12 pages.
3. Papers should be headed by a title, the initial(s) and surname(s) of
author(s) followed by address.
4. Each paper should bear abstract, 2 to 5 keywords, introduction,
methodology, observations, results and discussion followed by
acknowledgements and references.
5. In all studies of plants or animals proper identification should be made as
to the materials used.
6. While submitting the paper(s) for publication, Author(s) should decode
the drugs specially in case of clinical studies.
7. Bibliographical references should be listed in alphabetical order of the
author at the end of the paper. Authors should be cited in the text only
by their surname(s) but their initial(s) should be shown in the bibliography.
8. References to periodicals should include the name(s) and initial(s) of
author(s), year of publication, title of the book, periodical, title of the
article, volume number (Arabic numerals), issue number where appropriate,
first and last page number. Reference to books should include name(s)
and initial(s) of the author(s), year of publication, exact title, name(s) of
publisher, place of publication, page number.
9. Reference should be cited in the text in parentheses by the name(s) of
author(s) followed by the year of publication, e.g. “(Jain,1991)” except
when the author’s name is part of the sentence, e.g. “Jain (1991) has
reported that.” If there are more than two authors it is in order to put “
et al.” after the first name, e.g., Khan et al., 1981.
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148Hippocratic Journal of Unani Medicine
10. Each table should be typed on a separate sheet of paper. Tables should
be numbered consequently in Arabic numerals e.g. “Table 1, Table 2”
etc., and attached to the end of the text. Tables should be provided with
headings and kept as simple as possible and should be referred to in the
text as “table 1” etc.
11. Figures (including photographic prints, line drawings on strong white or
transparent paper, and maps) should be numbered consequently in Arabic
numerals, e.g. “Fig. 1 etc.” and attached to the text behind the tables.
Graphs and diagrams should be large enough to permit reduction to a
required size, legends for figures should be listed consequently on a
separate sheet of paper. Photographs should be on glossy printing paper.
12. The editors reserve the right to refuse any manuscript submitted, whether
on invitation or otherwise, and to make suggestions and modifications
before publication.
13. Paper accepted by the editorial board will become the property of the
CCRUM. No article or any part thereof may be reproduced in whatever
form, without the written permission of the Editor-in-Chief.
14. The editors and publisher are not responsible for the scientific contents
and statements of the authors of accepted papers.