PART
Solid Waste Management
C H A P T E R 2
, ciil Studies oil jh, ^ cnols isolated *'•'•' ^'irtjieric spent: An a^io i/idiistriul ask:
Chapter II Bioanalytical agro industrial waste
J4£stract
Extracts rf tut menc (Cuicuntci lri^£^a) spent \v£r£ pt£p.?>£^^ ix refluKii^tsi th£
tunuenc sv£ptt with £thuiisi.c£t:^t£. \vat£K kK£t/:anri a^^^ c>cetri^£. •"••tyrui^ui-u. a^Hru.yit
cf trli^t/!£i'vrl.s in tun'-u.£nc sp£i'Vt eKtuict (T.SE) was ritaLn£i^ •^'tri'i^- £ttiutac£tat£.
Trtal phenrUc crnt£nts ivete ^£t£miLn£y( USLI/I^I FrUn anal CLrcait£u's i'U£t/:ra(.
AntLCKidant activLtia .rf T^E was assaui£at throu^^h Ln v-Urr mralels such as p££
iadicai sca\/£naHn(r' acti^Lti-i ustnsi Q,:::-a(Lph£i-vud-i'VLCtujihufJiazuiL (DPPH)
methrd, fi-carpt£n£'LLnri£ate yAral£l S!^st£^H ana/ antirxcafant cavacitia bu Lrcn(!/f)
r£ductLrn m£thr;^~. AaldLttrn r^'' in.''^^^ ppkPi rf T:SE tr /£'7n£j sunflrw£y rtl and
strraaie at taring tet'Hp£iatut£ i2Z: ± 3'C) fat S^- a/auis shcw£ai Lr\v£> v£yrKLa(£ valu£
(Pov) (113.9'^ m£q rKuc*£n "iza) and ft££ fattia aad crntent (FFA)
{1.11 m'l kiOH 'a } criMpaied tr cmtirl sampL£s POv iii'i.5^ ^^^^ rKia^£n 'iza>)
and FFA (1.2^ mri KOH d). Riefined Sunflrwer rd crntaLninC* lOO ppm rf
butiaiatid huidirKL'ttrui£n£ (T^HT) and id-l^r pp^A. c^ -a*t££n tea prliat-h£nrl (^iFP)
and id'dd ppku. r-f rrseyvian.1 shrived PO\' r-f s.5.r:-d: ii2.r:^''^ and lO'^.^r: t'nea
rKuir^en 'tzp and FFA crntent r-^dS'^, l.r2, i.de- mr^ i<OH d, tespectL\,elia a-^er SO
daMs rf stnaae at rrry^ ten^perature. Fhe minimui^A cmcenttatims r-^ FSE
teauired fr> LnhLbitirn rf Escherichia crli, staphuitrcrccus auteus. salntrnelLa
tLuvhi, and^alnimeUa entenca '.'.£r£ S'^r. 1200, iiod andi3dd tt,A. tespectivelia.
kteui'-vrt ds: Cuicuipia Irnpa: tui i'U.inz stf^t e^ti .l:t: Antioxidant astivitu; Anr.r^i-i rii^l act:\,itu:
{tnr^-ins^usti'U ivastf
Jrurnal r^FrrdAr'ticultui-e and EniirrnrPi£nt (Crmmunicated)
Chapter II Bioanalytical agro industrial waste
II. 1.Introduction
Turmeric rhizome {Curcuma longa, family Zingiberaceae) is a well-
established foodstuff. It is a main ingredient of curry powders. It is also well known
for its coloring, flavoring, curative and digestive properties. Tiumeric is a well-known
indigenous herbal medicine, which exhibits a wide range of biological activities, for
example, antibacterial [1], anticancer [2], anticoagulant [3], antiinflammatory [4,5],
antimutagenic [5,6], antioxidant [6,7], antiprotozoan [8], antispasmodic [4],
antitumour [9,10], antiviral [5], hepatoprotective [5], hypocholesterolaemic [11],
hypoglycaemic [12], hypolipemic [13], besides being effective oxygen species
scavengers and lipid peroxidase inhibitor [14].
The active components of turmeric is yellow pigment curcumin (0.5 - 6.0%).
The minor constituents are suggested to be geometrical isomers of the major
constituent [15] collectively called curcuminoids, which are extracted for its
nutraceutical and cosmaceutical preparations and the process produces a large portion
of turmeric spent which is considered as waste. As most of the food industries are
localized the waste generated may contribute to environmental pollution. During past
few years, there has been increasing interest in isolating and recovering important
components from food industrial waste to add value-addition to the residues. The
emergence of the waste management for sustainable development by encouraging
controlled exploitation of food industrial waste for economic gains will help to add
value-addition to the turmeric spent.
Turmeric is native of south and south East Asia. Turmeric probably originated
in the slopes of hills in the tropical forest of western ghatts of south India. Turmeric
can be grown in diverse tropical environment from sea level to height of 1500 mts in
hill slopes in temperature ranging from 20 to 30°C. A rainfall of 150 cm/annum or
more in the growing region, or an equivalent amount of irrigation is essential. India
remains the largest producer of turmeric, its average and production exceeding the
total of all other growing countries together [16].
Antioxidants are compounds that help to inhibit many oxidation reactions
caused by free radicals. Free radicals are imstable molecules which include hydrogen
atom, nitric oxide and molecular oxygen. They naturally occur in the body as a result
of chemical reaction during normal cellular processes. Reactive oxygen species
21
Chapter II Bioanalytical agro industrial waste
(ROS) sometimes called active oxygen species are various forms of activated oxygen,
which include free radicals such as superoxide (O2') and hydroxyl radical (OH), as
well as non free radical species such as hydrogen peroxide (H2O2) [17].
Spoilage of food due to the presence of bacterial and fungal infection has been
a major concern for decades and it causes a considerable loss world wide. The
demand for non toxic, natural antibacterials has been rising with increased awareness
[18,19].
The objective of this investigation was to determine the total phenolic content
of the turmeric spent resulting from its extraction and to evaluate the use of
polyphenols as a potential source of natural antioxidant and antimicrobial for
commercial applications.
II.2. Materials and methods
11.2.1. Chemicals
Methanol, ethyl acetate, ethanol, ethyl ether, sulphuric acid, acetic acid,
chloroform, Folin and Ciocalteu's reagent, sodium carbonate were from Ranbaxy,
India; gallic acid, 2,2-diphenyl-l-picrylhydrazyl (DPPH), butylated hydroxytoulene
(BHT), potassium iodide, potassium hydroxide, P-carotene, linolenic acid, Tween 40
were from Sigma Chemicals, USA; potassium ferric cyanide, trichloroacetic acid,
sodium phosphate, ammonium molybdate were from BDH, India; nutrient agar was
from HiMedia, Mumbai. Green tea polyphenols (GTP) and rosemary were from
Flavors and Essence Ltd. Mysore, India. Refined sunflower oil (Sun pure brand) was
procured from the local market.
11.2.2. Instrumentation
Visible spectra measurements were done using Specord 50 UV-Vis
spectrophotometer with 1.0-cm silica quartz matched cell.
11.2.3. Plant material
The residue after exhaustive extraction of oleioresin from turmeric rhizomes
were collected from food industry (Flavors and Essence Pvt. Ltd.,) Mysore, India.
The residue is termed as turmeric spent.
22
Chapter II Bioanalytical agro industrial waste
11.2.4. Sample preparation
The turmeric spent was dried in hot air oven at 60°C to remove moisture and
ground into fine powder. The material that passed through a 40-mesh sieve was
retained for further studies.
77.2.5. Preparation of extract
Fifty grams of turmeric spent prepared as above was transferred to a 1000 ml
round bottom flask, 200 ml of ethylacetate was added and refluxed for 45 min at 60''C
on a water bath. The extract was filtered through filter paper and the residue was
reextracted under the same conditions five times. The filtrate was combined and
evaporated in a rotary evaporator below 60''C. The procedure was repeated using
solvents like acetone, methanol and water. The extract obtained after evaporation of
organic solvents was analyzed for percentage of polyphenols.
77.2.6. Determination of total phenolic content
Phenolic compounds are very important plant constituent because of their
scavenging ability due to their hydroxyl groups. According to the latest report, a
highly positive relationship between total phenolic compounds and antioxidant
activity was found in many plant species [20], Phenolic compounds were associated
with antioxidant activity and play an important role in stabilizing lipid peroxidation.
Thus, first-ever attempt has been made to isolate the phenolic contents of turmeric
spent to provide value-addition to the food industrial residue.
The phenolic components were determined according to the procedure
described by Negi (2003) [21]. Twenty mg of extract were dissolved in 50 ml of 50 %
(v/v) methanol and filtered through Whatman no 44 filter paper. An aliquot of 0.4 ml
of sample was taken in a 25-ml standard flask and mixed with 1 ml of Folin and
Ciocalteu's reagent (1:10 diluted with water) and allowed to react for 5 min, 1 ml of
10 % sodium carbonate was added and allowed to stand for 90 min at room
temperature and the absorbance of the reaction mixture was read at 725 nm. Standard
graph was prepared using gallic acid as reference. The content of total polyphenols in
the extract was calculated firom the standard graph and expressed as percent
concentration in the extract.
23
Chapter 11 Bioanalytical agro industrial waste
II.2.7. Quenching of 2,2-diphenyl-l-picrylhydrazyl (DPPH) radical by TSE
The method described by Kitts [22] was used to assess the DPPH radical
scavenging capacity of TSE. A 0.1 mM DPPH solution in ethanol was mixed with
20, 40, 60, 80, 100, 120, 140 ng/ml of TSE and vortexed thoroughly. The mixtures
were allowed to stand at ambient temperature (22 ± 3°C) for 30 min. The absorbance
was measured at 519 nm using a spectrophotometer. The scavenging percentage was
calculated as per the following equation.
C . 0 / ( A b Control - A b sample ) X 1 0 0
Scavengmg, % = A " Control
II.2.8. Antioxidant assay using fi-carotene-linoleate model system
The antioxidant activity of TSE was evaluated using p-carotene-linoleate
model system as described by Jayaprakasha [23]. P-carotene (0.2 mg) in 0.2 ml of
chloroform, 20 mg of linolenic acid and 200 mg of Tween-40 (polyoxyethylene
sorbitan monopalmitate) were mixed. Chloroform was removed at 40*'C under
vacuum and the resulting mixture was diluted with 10 ml of water and mixed well. To
this 40 ml of oxygenated water was added. Four milliliter aliquots of the emulsion
were pipetted into different test tubes containing 0.2 ml TSE (200, 400, 600, 800 ^g)
and BHT (200, 400, 600, 800 ng) in ethanol. BHT was used for the comparative
study. Control contained 0.2 ml of ethanol and 4 ml of the above emulsion was
prepared. The tubes were placed in 50°C in water bath and the absorbance at 470 rmi
was taken at zero time (t = 0 min). Measurement of absorbance was continued till at
an interval of 15 min the color of P-carotene disappeared in the control tubes (t =120
min). A mixture prepared as above without P-carotene served as blank. All
determinations were carried out in triplicates. The antioxidant activity (AA) of the
extract was evaluated based on the extent of bleaching of the P-carotene using the
formula.
100[1-(A„-A.)] (A°o-A«t)
24
Chapter II Bioanalytical agro industrial waste
Where, Ao and Ao* are the absorbance values measured at zero time of the
incubation for test sample and control, respectively. At and A 't are the absorbance
measured in test sample and control respectively after incubation for 120 min.
11.2.9. Reducing power ofTSE
The reducing power of TSE was determined following the method of
Oyaizu [24]. Different amounts of TSE extract (0.00, 0.24, 0.60, 1.20, 1.80, 2.40,
3.00 mg/ml) in water were mixed with phosphate buffer (2.5 ml, 0.2 M, pH 6.6) and
potassium ferricyanide [K3Fe(CN)6] (2.5 ml, 1%). The mixture was incubated at 50°C
for 20 min. Trichloroacetic acid (10%, 2.5 ml) was added to the mixture and
centrifuged for 10 min. The upper layer of the solution (2.5 ml) was mixed with
distilled water (2.5 ml) and FeCb (0.5 ml, 0.1 %), and the absorbance was measured
at 700 nm. Absorbance of the reaction mixture increased with the increase in reducing
power.
11.2.10. Antioxidant activity assay using sunflower oil
Refined sunflower oil, fi"ee of additives was used as the substrate for oxidation
studies. Sunflower oil samples containing 1000 ppm of TSE and natural antioxidants
(GTP and rosemary) were separately prepared. Each 250 ml prepared oil sample was
placed in a 500 ml brown airtight glass bottle. Synthetic antioxidant (BHT) was also
mixed at its permitted limit of 100 ppm with sunflower oil for comparison [25].
Control sample of sunflower oil without the addition of antioxidant was stored under
identical conditions. All oil samples in each treatment were prepared in triplicates and
were stored at room temperature (22 ± 3°C) for 80 days. Oil sample from each
treatment was withdrawn periodically to assess the extent of oxidation.
II.2.10.1. Antioxidant activity measurement
The rate of oxidation was followed by periodic determination of POV of
stored sunflower oil subjected to different treatments. Each sample (1 g) was taken in
250-ml Erlenmeyer flask and dissolved in a mixture of 25 ml of acetic
acid/chloroform (3:2, v/v). Saturated solution of potassium iodide (1 ml) was added
and the flask was placed in a dark chamber for 5 min, after which 75 ml distilled
25
Chapter 11 Bioanafytical agro industrial waste
water was added. The liberated iodine was titrated against a sodium thiosulfate
solution (0.01 M) in the presence of starch indicator [26].
II.2.10.2. Free fatty acid content
Free fatty acid content was determined at regular intervals in stored sunflower
oil. 10 grams of each sample were weighed into an Erlermieyer flask and 50 ml of
ethanol/ethyl ether (1:1, v/v) mixture were added. This suspension was titrated
against O.IM potassium hydroxide (KOH), using 1 ml of phenolpthalein indicator,
until faint permanent pink color persisting for 30 Sec was obtained [26].
II.2.11. Antibacterial activity
Bacterial cultures of Escherichia coli. Staphylococcus aureus. Salmonella
typhi and Salmonella enterica were obtained from Defence Food Research
Laboratory, Mysore, India. The cultures were grown in nutrient agar at 37°C. Each
bacterial strain was transferred from slants at 4 -5°C to 10 ml broth and cultivated
overnight at 2)TC. A preculture was prepared by transferring 1 ml of this culture to
9 ml broth and cultivated for 48 h. The cells were harvested by centrifiigation
(1200 g, 5 min), washed and suspended in saline.
The TSE was tested against the growth of bacteria by the method of Negi
(1999)[27]. To flasks containing 20 ml melted cool agar, different concentrations of
test material in Tween 40 were added. Equivalent amounts of Tween 40 were used as
controls. One hundred microliter (about 10 cfii/ml) of each bacterium to be tested
were inoculated into the flasks imder aseptic conditions. The media were then poured
into sterilized petri plates in duplicate and incubated at 37°C for 20-24 h. The
colonies, after the incubation period were counted and expressed as colony forming
units per ml (cfii/ml) of culture. The minimum inhibitory concenfration (MIC) was
determined as the lowest concentration of the TSE required for inhibiting the
complete growth of the bacterium being tested.
26
Chapter II Bioanalytical agro industrial waste
II.3. Results and discussion
The % yields of TSE in ethyl acetate, acetone, methanol and water were 5.25,
7.13, 7.76 and 13.3 respectively. The total content of phenolics in TSE as determined
by Folin-Ciocalteu method are reported as gallic acid equivalents. Amongst the
various solvents tried ethylacetate gave the highest polyphenolic content. The order of
the efficiency of the different solvents in enhancing polyphenolics from TSE was
ethylacetate (6.75%) > acetone (4.11%) > methanol (3.91%)>water (3.12%).
The total phenolic contents in TSE determined are not absolute values but are
based on their chemical reducing capacity relative to gallic acid. In the present study
the responses of the TSE may arise from the variety and/or quantity of phenolics
found in extracts of turmeric spent.
Table II.l. Percent radical-scavenging activity on DPPH by TSE and gallic acid
Extract
concentration
20
40
60
80
100
120
140
TSE
(% scavenging)
20.36 ±0.37
37.4710.71
54.56 ±0.55
67.70 ± 0.67
78.31 ±0.48
82.81 ±0.77
84.71 ±0.28
Gallic acid
(% scavenging)
92.71 ±0.14
93.81 ±0.28
94.11 ±0.22
94.71 ±0.31
94.86 ± 0.48
94.46 ± 0.33
94.81 ±0.19
Mean ± Standard deviations (n=3)
II. 3.1. Free radical scavenging activity of TSE
The scavenging activity of TSE phenolics compared to gallic acid for DPPH
radical is shown in Table II.l. The results indicate a concentration-dependent
scavenging activity of the DPPH radical. Further, it was found that the scavenging
activity of TSE was found to be maximum at 140 |xg/ml and it was 84.71 %.
27
Chapter II Bioanalytical agro industrial waste
n.3.2. Antioxidant activity using fi-carotene-linoleate model system
Antioxidant activity of TSE in P-carotene-Iinoleate model system is shown in
Fig ILL Addition of TSE and BHT prevented the bleaching of P-carotene but to
different degrees. P-Carotene undergoes rapid discoloration in the absence of an
antioxidant. This is due to the coupled oxidation of P-carotene and linolenic acid,
which generate free radicals. The linolenic free radicals formed upon the abstraction
of a hydrogen atom from one of its diallylicmethylene groups attacks the highly
unsaturated P-carotene molecules. As a result, p-carotene gets oxidized and broken
down in part, subsequently the system looses its chromophore and characteristic
orange color; this can be monitored spectrophotometrically. In the present study, the
TSE was found to hinder bleaching of P-carotene by neutralizing the linoleate free
radical and other free radicals formed in the system. TSE showed a maximum of
92.74 % antioxidant activity when used at the highest concentration of 800 \x%.
120
SJ-^ > ^^ o 4->
C m
•D X o
4 ^
c <
100
80
60
40
?n
0
USE I BHT
200 400 600
Concentration (pg)
800
Fig.II.l. Antioxidant activity of extract from TSE and BHT by
p-carotene- linoleate model system
28
Chapter 11 Bioanalytkal agro industrial waste
11.3.3. Reducing power
Fig II.2 shows the reducing capacity of TSE and it is compared with BHT. For
this, we investigated the Fe /Fe transformation in the presence of TSE, using the
method of Oyaizu (1986)[24]. Earlier authors [28,29] have observed a direct
correlation between antioxidant activity and reducing power of certain plant extracts.
The reducing properties are generally associated with the presence of reductones, [28]
which have been shown to exert antioxidant action by breaking the free radical chain
by donating a hydrogen atom [30]. Reductones also react v^th certain precursors of
peroxide, thus preventing peroxide formation, hi Fig II.2 the reducing power of TSE
suggests that it contribute significantly towards the antioxidant effect. The reducing
power of TSE is closely related to the amount of the extract being used and increases
with increasing amovmt of the extract. However, the reducing power of BHT was
relatively greater than that of TSE.
1.6.
1.4-
r; S0.8
1 0 . 6
5 0.4
0.2 n)
( F 1 1 1 1
) 1 2 3 4
Concentration
-•—Series 1
-»-Series2
Fig.II.2. Reducing power of turmeric spent extract at different concentrations
II.3.4. Antioxidant activity of TSE in refined sunflower oil
Oxidation of lipids is imdesirable as it affects the flavor, aroma, nutritional
quality and even the texture of the product where it is used. The chemicals produced
by the oxidation of lipids are responsible for rancid flavor and aroma.
29
Chapter II Bioanalytical agro industrial waste
^ 180 1 o> ^ 160 a> w 140 -X f 120-
E 100-•^ 80-o •i 60 ra > « 40
•a
g 20 W
g. 0^ (
/ ,_^,-r^
1 1 1 1 1
) 20 40 60 80 100
Storage period (Days)
—•—Control
- * - B H T
GTP
—tr- Rosemary
-JK-TSE
Fig.II.3. Effect of synthetic antioxidant BHT (100 ppm), natural antioxidants
GTP (1000 ppm) and rosemary (1000 ppm) and TSE (1000 ppm) on
peroxide value (mmol oxygen /kg) during storage of sun flower
oil
1.4-
1.2
" SO.8
150-6 u. 0)0.4
^0.2
n -
( ) 20 40 60 80 100
Storage period (days)
—•—Control
GTP
K Rosemary
-)is-TSE
Fig.II.4. Effect of synthetic antioxidant BHT (100 ppm), natural
antioxidants GTP (1000 ppm), and rosemary (1000 ppm) and
TSE (1000 ppm) on free fatty acid (mg KOH/g sample) during
storage of sim flower oil.
30
Chapter II Bioanalytical agro industrial waste
BHT, GTP and rosemary are commonly used as antioxidants in oils. BHT is a
synthetic while; GTP and rosemary are natural antioxidants. The antioxidant property
of TSE was determined and compared with BHT, GTP and rosemary. The result
presented in Fig II.3 and II.4 manifest that TSE has antioxidant property, which is
comparable to GTP and rosemary. But TSE has lesser antioxidant activity when
compared to BHT. The FFA and POV did not increase much, which clearly indicates
that autooxidation of sunflower oil was greatly inhibited by the TSE. This study
revealed that the TSE can be used as a natural antioxidant at little higher
concentration compared to natural antioxidants like GTP and rosemary.
11.3.5. Antimicrobial activity
2000
1600-Minimum Inhibitory 1200
Concentration „ . . (ppm)
400
0 1^^ *
^ • 1
^v
<
-1 1 1 1
1 2 3 4 Bacteria
Fig. II.5. Effect of TSE on growth of Escherichia coli (1),
Staphylococcus aureus (2), Salmonella typhi (3) and
Salmonella enterica (4).
The effect of TSE on the growth of four different bacteria is dose dependent
as presented in Fig II.5. TSE inhibited growth of bacteria to variable extent,
depending on the bacterium in question. TSE completely inhibited the growth of
E.coli at 850 ppm concentration. But S. aureus, S. typhi and S.enterica required 1200,
1100, 1300 ppm of concentration respectively for the complete inhibition of growth.
31
Chapter 11 Bioanalytkal agro industrial waste
II.4. Conclusion
First-ever studies on turmeric spent have been carried out to recover the
polyphenols, which have demonstrated antioxidant and antimicrobial properties. The
study has been carried out to develop natural and cost-effective antioxidant material
from solid waste generated by food-industry. The extract from turmeric spent has
indicated high antioxidant and antibacterial activities. Further studies are in progress
to characterize individual phenolics and to elucidate the mechanism underlying
bioactive properties and existence of possible synergism, if any, among these
compounds.
In summary, the varied experience in industrial and agricultural sectors amply
proves that the crucial importance in management is in the realization of the benefits
of development in an optimum and sustainable manner. The problem is more acute
and wide spread in developing economics coupled with pressing social priorities.
Such problem acquires serious magnitude in polluting the environment. In such a
situation engineering of food-industrial waste for feed, food and functional foods
assumes paramount importance. This paper is a step in this direction.
32
Chapter II Bioanafytical agro industrial waste
Refernces
1. J. Lutomski, B. Jedzia and W. Debska, "Effect of an alcohol extract and of active ingredients
from Curcuma longa on bacteria and fimgi", Planta Med., 26 (1974) 9-19.
2. H.W. Chen and H.C. Huang, "Effect of curcumin on cell cycle progression and apoptosis in
vascular smooth cells", Br. J. Pharmacol., 124 (1998) 1029-1040.
3. R. Srivastava, M. Dikshit, R, C. Srimal and B.N. Dhawan, "Anti thrombotic effect of
curcumin", Thromb. Res., 40 (1985) 413-417.
4. H.P.T Ammon and M.A. Wahl, "Pharmacology of Curcuma longa", Planta Med., 57 (1991)
1-7.
5. R.C. Srimal, "Turmeric: A brief review of medicinal properties", Fitoterapia., 68 (1997) 483-
493.
6. 6.1. Ross, "Medicinal Herbs of the World", Humana Press, Totowa, New Jersey, 1 (1999).
7. 7. D.S. Kim, S.Y. Park and J.K. Kin, "Curcuminoids from Curcuma longa L. (Zingiberaceae)
that protect PC 12 rat pheochromocytoma and normal human umbilical vein endothelial cells
from beta-amyloid (1—^> 42) insult", Neurosci. Lett., 303 (2001) 57-61.
8. T.N. Bhavani Shankar and V. Sreenivasa Murthy, "Effect of turmeric (Curcuma longa)
fractions on the growth of some intestinal and pathogenic bacteria in vitro", Indian J. Exp.
Biol., 17(1979)1363-1366.
9. Y.C. Chen, T.C. Kuo, S.Y. Lin-Shiau and J.K. Lin, "Induction of HSP70 gene expression by
modulation of Ca2+ ion and cellular P53 protein by curcumin in colorectal carcinoma cells',
Mol. Carcinogenesis, 17 (1996) 224-234.
10. D. Deeb, Y. X. Xu, H. Jiang, X. Gao, N. Janakiram, R.A. Chapman and S.C. Gautam,
"Ciu-cumin (diferuloyl-methane) enhances tumor necrosis factor-related apoptosis-induced
apoptosis in LNCaP prostate cancer cells", Mol. Cancer Ther., 2 (2003) 95-103.
11. R.C. Srimal and B.N. Dhawan, "Pharmacology of diferulogyl methane (ciu-cumin), a non
steroidal anti-inflammatory agent", J. Pharm. Pharmacol., 25 (1973) 447-452.
12. J. Blasiak, A. Trzeciak, E. Malecka-Panas, J. Drzewoski, T. Iwamienkoo, I. Szumie and M.
Wojewodzka, "DNA damage and repak in human lymphocytes and gastric mucosa cells
exposed to chromium and curcumm", Teratogen. Carcinogen. Mutagen., 19 (1999) 19-31.
13. S.D. Rao, N. Chandrashekhara, M.N. Satyanarayana and M.Srmivasan, "Effect of curcumin
on serum and liver cholesterol levels in the rat", J. Nufr., 100 (1970)1307-1315.
33
Chapter II Bioanalytical agro Industrial waste
14. E. Kunchandy and M.N.A Rao, "Oxygen radical scavenging activity of curcumin", Intl. J.
Pharm., 38 (1990) 239-240.
15. K.R. Srinivasan, "The coloring matter in turmeric", Curr. Sci., (1952) 311-312.
16. T.T. Panlose and E. Velappan, "Future prospects for spice industry", Proc. Symp.
Development prospects of spice industry in India. Assoc. Food Sci. Tech (india) (1974) 78.
17. Duthie, G.G., Duthie, S.J. and Kyle, J.A.M, "Plant polyphenols in cancer and heart disease:
implications as nutritional antioxidants". Nutrition research reviews, 13, (2000) 79-106.
18. Davies J, "Inactivation of antibiotics and dissemination of resistant genes. Science, 264
(1994) 375-382
19. Enne, V. I., More, D.M. L., Stephens, P. and Hall, L. M. C, "Persistance of sulphonamide
resistance in Escherichia coli in the U.K. despite national prescribing restriction". Lancet, 28
(2001) 1325-1328.
20. Negi, P.S, Chauhan, Sadia, Rohinishree and Ramteke, Food Chemistry (2005).
21. Negi, P.S., Jayaprakasha, G.K. and Jena, B.S, "Antioxidant and antimutagenic activities of
pomegranate peel extracts". Food Chemistry, 80 (2003) 393-397.
22. Kitts, D.D., Wijewickreme, A.N. and Hu, C, "Antioxidant properties of a North American
Ginseng extract". Molecular and Cellular Biochemistry, 203 (2000) 1-10.
23. Jayaprakasha. G.K., Singh, R.P. and Sakiariah, K.K, "Antioxidant activity of grape seed
(Vitis vinifera)". Food Chemistry, 73 (2001) 285-290.
24. Oyaizu M, "Studies on product of browning reaction prepared from glucose amine:" Japanese
Journal of Nutrition, 44 (1986) 307-315.
25. Duh, P.D. and Yen, G.C, "antioxidant efficacy of methanolic extracts of peanut hulls in
soybean and peanut oils"JoumaI of American Oil Chemical Society, 74 (1997) 45-748.
26. AOCS Official method of analysis (15th ed.). Washington, DC: Association of Official
Analytical Chemists (1990).
27. Negi, P.S. Jayaprakasha, G.K., Rao, L.J.M. and Sakariah, K.K, "antibacterial activity of
turmeric oil-a byproduct from Curcumin manufacture" Journal of Agricultural and Food
Chemistry, 47 (1999) 297-4300.
28. Pin- Der-Dhu, X,Antioxidant activity of burdock (Arctium lappa Linne): its scavenging effect
on free-radical and active oxygen",Journal of American Oil Chemists Society. 75 (1998) 5-
461.
34
Chapter II Bioanalytical agro industrial waste
29. Pin-Der-Duh. X., Pin-Chan-Du. X. and Gow-Chin Yen. X. Action of methanolic extract of
mung hulls as inhibitors of lipid peroxidation and non-lipid oxidative damage",Food and
Chemical Toxicology. 37, (1999)1055-1061.
30. Gordon, M. H. The mechanism of antioxidant action in vitro. In B. J. F. Hudson (Ed.), Food
antioxidants (pp. 1-18). London: Elsevier Applied Science (1990).
35
Top Related