In Vitro Pharmacological activity of Zinc and Copper ...joics.org/gallery/ics-3567-.pdf · In Vitro...
Transcript of In Vitro Pharmacological activity of Zinc and Copper ...joics.org/gallery/ics-3567-.pdf · In Vitro...
In Vitro Pharmacological activity of Zinc and Copper
nanoparticles using medicinal plant of Acalypha indica
root extracts.
N. Muruganantham*1 and P. Anitha2
*1 PG & Research Department of Chemistry, Thanthai Hans Roever College (Autonomous),
(Affiliated to Bharathidasan University), Perambalur - 621220, Tamil Nadu, India. 2Department of Physics, Roever College of Engineering and Technology, (Affiliated to Anna
University), Perambalur - 621220, Tamil Nadu, India.
*Corresponding author: [email protected]
Abstract
Zinc and Copper nanoparticles are synthesized from aqueous extracts of Acalypha indica root.
Pharmacological activitis such as Anti-oxidant, Anti- inflammatory activity and Anti diabetic
activity were studied. The ZnNPs and CuNPs have showed significant antibacterial activity on
multi drug resistance in biological fields. The ZnNPs and CuNPs had shown strong antioxidant
by DPPH scavenging activity. The ZnNPs and CuNPs exhibited strong anti-inflammatory
activity by albumin denaturation activity. The ZnNPs and CuNPs had strongly inhibited the
aglucosidase to a-amylase. To the best of our knowledge, this is the first attempt on the
synthesis of ZnNPs and CuNPs nanoparticles using Acalypha indica root extracts. Hence, to
validate our results, the in vivo studies at molecular level are needed to develop an antioxidant,
anti-diabetic and anti-inflammatory agent. Results of the present study, highlighted the eco-
friendly approach of plant mediated synthesis of nano particles and its potential application in the
field as an alternative to chemical drug for disease management.
Keywords: Zinc and Copper nanoparticles, antioxidant, anti-diabetic and anti-inflammatory
agent etc.,
1. Introduction
Medicinal herbs are moving from fringe to mainstream use with a greater number of
people seeking remedies and health approaches free from side effects caused by synthetic
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org631
chemicals. India officially recognizes over 3000 plants for their medicinal value. It is generally
estimated that over 6000 plants in India are in use in traditional, folk and herbal medicine. This
article aims to provide a comprehensive review on the pharmacological aspects of Acalypha
indica. It is obtained from deciduous and mixed-monsoon forests throughout greater parts of
India.
Over the last few years, researchers have aimed at identifying and validating plant-
derived substances for the treatment of various diseases. Interestingly it is estimated that more
than 25% of the modern medicines are directly or indirectly derived from plants. It is worth
mentioning that Indian medicinal plants are considered as a vast source of several
pharmacologically principles and compounds that are commonly used as home remedies against
multiple ailments [1].
Indian traditional medicine is based on various systems including Ayurveda, Siddha,
Unani and Homoeopathy. The evaluation of these drugs is primarily based on phytochemical,
pharmacological and allied approaches including various instrumental techniques such
chromatography, microscopy and others. With the emerging worldwide interest in adopting and
studying traditional systems and exploiting their potential based on different health care systems,
the evaluation of the rich heritage of traditional medicine is essential [2].
In this regard, one such plant is Acalypha indica L. Acalypha indica has been extensively
used in Ayurvedic system of medicine for various ailments. It is deciduous and mixed-monsoon
forests throughout greater parts of India, is widely used in traditional medicinal system of India
has been reported to possess hepatoprotective, anti-inflammatory, antitussive, antifungal and
used also check wounds healing and antibacterial [2].
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org632
Taxonomic Classification
Fig.1 Acalypha indica Fig.2 Acalypha indica
(Root)
Table.1 Taxonomic Classification
Among all the noble metal nano particles, silver nanoparticles have gained interests
because of their unique properties such as chemical stability, good conductivity, catalytic and
most important antibacterial, antiviral, antifungal and anti-inflammatory activities [3]. The wide
applications of silver nano particles has opened up another arena of research interest that is green
nanotechnology or biosynthesis of nanoparticles utilizing algae, actinomycetes, bacteria, fungi
and plants. Among the various biological synthesis procedures plant mediated synthesis of silver
nanoparticles has attracted more attention nowadays. Synthesis of silver nanoparticles has been
investigated utilizing many plant extracts like Marigold flowers [4], leaf extract of Acalypha
indica [5]
The silver nanoparticles are the most concentrated interest for research due to its potential
in applications like to transform the physical, electronic and optical properties of a compound,
treatment of cancer, as medical devices, antimicrobial coatings of paint, textiles, also biological
Kingdom : Plantae
Class : Magnoliopsida
Order : Euphorbiales
Family : Euphorbiaceae
Subfamily : Acalyphoideae
Genus : Acalypha
Species : Acalypha indica Linn.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org633
or chemical sensing, also possess anti inflammatory, antiviral, antifungal and antiplatelet
activities [6,7]. In this present study, a plant extract of Acalypha indica are used, which grows in
wastelands, gardens, and roadside. They are herbaceous annual weed that grows up to a height of
15 m. It falls under the family of Euphorbiaceae, which has diuretic, antimicrobial, anti-
helminthic properties and used in the treatment of respiratory problems like bronchitis,
pneumonia, and asthma [8].
This plant is used as diuretic, antihelmintic and for respiratory problems such as
bronchitis, asthma and pneumonia [9]. The roots of A. indica is used as laxative and leaves for
scabies and other cutaneous diseases [10]. Major phytochemicals identified from A. indica are
acalyphine, cyanogenic glycoside, inositol, resin, triacetomamine and volatile oils [11]. This
plant has been used extensively in herbal medicine in many tropical and sub tropical countries
[12,13]. A. indica having analgesic and anti-inflammatory effects. In Malaysia, A. indica is used
for generations for the treatment of superficial fungal and several other bacterial infections
[14].Previous studies on A. indica revealed that this plant has antibacterial activity against
several gram positive bacteria [15,16].
Gold nanoparticles of size 20-30 nm were rapidly synthesized using aqueous leaves
extract of Acalypha indica as novel source of bio-reductants [17]. Incidentally, biological
systems have long been known to reduce metal ions into nano particles [18] and many
researchers have recently reported the biogenic synthesis of silver and gold nano size
particles using a wide range of biological resources like bacteria [19]fungi [20]and plants [21]n
the plant mediated green chemistry approach, the reduction rate of metal salts is very fast and the
procedure itself requires no specific conditions unlike the physical and chemical methods [22,23]
Besides, this biogenic method of nanoparticles are synthesis appears to be reproducible and the
particles, produced through this environmentally friendly approach, are found highly stable [24-
26].
In the study, the silver nanoparticles were synthesized from these plant extracts, and
checked for their Anti-oxidant, Anti- inflammatory activity and Anti diabetic activity.
2. Materials and methods.
2.1 Collection of root
Fresh root of the samples were collected from Perambalur, during the month of December.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org634
2.2 Preparation of root extract
The fresh and young root of the samples were collected & washed thoroughly with sterile double
distilled water (DDW). Twenty grams of sterilized root samples were taken and cut into small
pieces. Finely cut roots were placed in a 500 ml Erlenmeyer flask containing 50ml of sterile
DDW. After that, the mixture was transferred to Soxhlet apparatus to derive extracts. The extract
was stored in 4 0C.
2.3 Microwave assisted synthesis of metal nanoparticles
Metal nitrate was used as precursor in the synthesis of metal nanoparticles. 100 ml of root extract
was added to 100 ml of 0.1N metal nitrate aqueous solution in conical flask of 250 ml content at
room temperature. The flask was thereafter put into shaker (100 rpm) at 500 C and reaction was
carried out for a period of 12 hrs. Then the mixture is kept in microwave oven for exposure of
heat.
Metals nanoparticles are made by a chemical reduction of a metal salt in the presence of a
stabilizing agent. Rapid microwave heating and agitation gives monodispersed particles. Add
200 ml of extract with 1M metal nitrate in beaker and Cover loosely. Expose the sample in
Microwave radiation for 20 minutes at 100% power. The setting of time is done on the basis of
trial and error method. The color will continue to change with respect to time. The mixture was
completely dried after a period of 20 minutes and hence nano particles in form of powders were
obtained.
2.4 Anti-Oxidant Studies
DPPH scavenging assay
The ability to scavenging the stable free radical, DPPH was measured as a decrease in
absorbance at 517 nm by the method.
Reagents
2,2-Diphenyl-1-picryl hydrazyl (DPPH) – 90.25mM in methanol in a dark room.
Procedure
To a methanolic solution of DPPH (90.25 mM), an equal volume of ethanolic Rhizome of
Cyperus rotundus L (250-1500 µg) was added and made up to 1.0 mL with methanolic DPPH.
An equal amount of methanol was added to the control. After 20 min, the absorbance was
recorded at 517 nm in a Systronics UV-visible Spectrophotometer. Ascorbic acid was used as
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org635
standard for comparison. The inhibition of free radicals by DPPH in percentage terms (%) was
calculated by using the following equation.
% Scavenging = (OD of Control- OD of Sample/ OD of Control) X 100.
Where A control is the absorbance of the control reaction (containing all reagents except the test
compound), and A sample is the absorbance of the test compound.
2.5 Anti- inflammatory activity
Inhibition of Albumen Denaturation
The reaction mixture was consisting of test extracts and 1% solution of bovine albumin fraction,
pH of the reaction mixture was adjusted using small amount at 37°C HCl. The sample extracts
were incubated at 37°C for 20 minutes and then heated to 51°C for 20 minutes after cooling the
samples the turbidity was measured spectrophotometrically at 660 nm. Diclofenac sodium was
taken as a standard drug. The experiment was performed in triplicates. Percent inhibition of
protein denaturation was calculated as follows:
Percent inhibition (%) = (OD of Control- OD of Sample/ OD of Control) X 100.
2.6 Inhibition Of Alpha-Amylase Enzyme
Starch solution (0.1% w/v) was prepared by stirring 0.1 g of potato starch in 100 ml of 16 mM of
sodium acetate buffer. The enzyme solution was prepared by mixing 27.5 mg of α-amylase in
100 ml of distilled water. The colorimetric reagent is prepared by mixing sodium potassium
tartarate solution and 3,5-di nitro salicylic acid solution 96 mM. The starch solution is added to
the both control and plants extract tubes and left to react with α-amylase solution, under alkaline
conditions at 25ºC. The reaction was allowed for 3 min. The generation of maltose was
quantified by the reduction of 3,5-dinitro salicylic acid to 3-amino-5-nitro salicylic acid. This
reaction is detectable at 540 nm.
% Inhibition = (OD of Control- OD of Sample/ OD of Control) X 100.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org636
3. Result and Discussion
3.1 Anti – Oxidant activity
DPPH scavenging assay method
There are several methods available to assess the antioxidant activity of compounds. DPPH free
radical scavenging assay is an easy, rapid, and sensitive method for the antioxidant screening of
plant extracts. In the presence of an antioxidant, DPPH radical obtains one more electron and the
absorbance decreases.
In the present study, the ZnNPs and CuNPs using root extracts of Acalypha indica high
DPPH scavenging capacity, which increased with increasing concentration [Table 2 and Figure
3]. It is evident from the data presented in Table, that the sample possesses DPPH assay activity.
For the ZnNPs, the result shows the percentage of cytotoxicity for 250 mg/ml as 10.09%, 500
mg/ml as 32% and 750 mg/ml as 49.48%. For the CuNPs, the result shows the percentage of
cytotoxicity for 250 mg/ml as 41.25%, 500 mg/ml as 51.91% and 750 mg/ml as 72.67%. These
inhibition values are compared with standard drug of Ascorbic acid for 250 mg/ml as 23.63%,
500 mg/ml as 29.00% and 750 mg/ml as 45.25%.
The DPPH assay was carried out at different concentrations of ZnNPs and CuNPs using
root extracts of Acalypha indica, namely 250 mg/ml, 500 mg/ml and 750 mg/ml. DPPH assay
did not show any significant difference at 250 mg/ml Concentrations in Acalypha indica,
however, it was significant for 500 mg/ml and 750 mg/ml for the nanoparticles, all the values are
compared with standard drug of Ascorbic acid. Hence, this assay provided information on the
reactivity of test samples with a stable free radical.
As a part of the investigation on the mechanism of the anti‐oxidant activity, ability of
extract to inhibit DPPH scavenging assay was studied. The in-vitro study of anti‐oxidant activity
indicates that the inhibition percentage of DPPH scavenging assay by Acalypha indica root
extracts of CuNPs activity is higher than ZnNPs.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org637
S.No Test
Concentration
of the sample
(mg/ml)
% of
inhibition of
the ZnNPs
% of
inhibition of
the CuNPs
Ascorbic
acid
(Standard)
1
DPPH
250 10.09 41.25 23.63
2 500 32 51.91 29.00
3 750 49.48 72.67 45.25
Table.2 Anti Oxidant activity of ZnNPs and CuNPs using root extracts of Acalypha indica
by DPPH Scavenging assay.
Fig.3 Graphical representation of Anti oxidant activity of ZnNPs and CuNPs using root
extracts of Acalypha indica by DPPH Scavenging assay.
3.2 Anti- inflammatory activity
Inhibition of Albumen Denaturation method
There are certain problems in using animals in experimental pharmacological research, such as
ethical issues and the lack of rationale for their use when other suitable methods are available.
Hence, in the present study, the protein denaturation bioassay was selected for in vitro
assessment of the anti-inflammatory property of Cobalt nanoparticles synthesized Acalypha
0
10
20
30
40
50
60
70
80
250 500 750
% o
f in
hib
itio
n
Concentration
DPPH Scavenging assay activity
% of inhibitionof the ZnNPs
% of inhibitionof the CuNPs
Ascorbic acid(Standard)
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org638
indica. The Albumen Denaturation is a well-documented cause of inflammation. Most biological
proteins lose their biological functions when denatured. Production of autoantigen in certain
arthritic disease is due to denaturation of protein. The mechanism of denaturation involves an
alteration in electrostatic hydrogen, hydrophobic, and disulfide bonding. In the present study,
denaturation of proteins is the main cause of inflammation. As part of the investigation on the
mechanism of the anti-inflammatory activity, ability of the extract to inhibit protein denaturation
was studied. Selected extracts were effective in inhibiting heat-induced albumin denaturation.
Aspirin was used as a standard anti-inflammation drug as shown in Figure [Table 3 and Figure
4]. The albumin denaturation method was carried out for ZnNPs and CuNps at different
concentrations such as 100µg/ml 200µg/ml and 300 µg/ml.
For the ZnNPs, the result shows the percentage of cytotoxicity for 100 mg/ml as 43.07%,
200 mg/ml as 52.20% and 300 mg/ml as 58.02%. For the CuNPs, the result shows the percentage
of cytotoxicity for 100 mg/ml as 48.04%, 200 mg/ml as 58.08% and 300 mg/ml as 64.32%.
These inhibition values are compared with standard drug of Aspirin
for 100 mg/ml as 45%, 200 mg/ml as 56.25% and 300 mg/ml as 66.20%. Albumen Denaturation
did not show any significant difference at 100 mg/ml Concentrations in Acalypha indica,
however, there was a significant change when the concentrations are 200 mg/ml and 300 mg/ml
for the nanoparticles and all the values are compared with standard drug of Aspirin.
As a part of the investigation on the mechanism of the anti‐oxidant activity, ability of
extract to inhibit Inhibition of Albumen Denaturation was studied. The in-vitro study of Anti-
nflammatory activity indicates that the inhibition percentage of Albumen Denaturation by
Acalypha indica root extracts of CuNPs activity is higher than ZnNPs.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org639
S.No Test
Concentration
of the sample
(mg/ml)
% of Protein
Denaturation
of the ZnNPs
% of Protein
Denaturation
of the CuNPs
Aspirin
(Standard)
1
Albumin
denaturation
100 43.07 48.04 45
2 200 52.20 58.08 56.25
3 300 58.02 64.32 66.20
Table.3 Anti-inflammatory activity of ZnNPs and CuNPs using root extracts of Acalypha
indica by Albumen Denaturation.
Fig.4 Graphical representation of Anti-inflammatory activity of ZnNPs and CuNPs using
root extracts of Acalypha indica by Albumen Denaturation.
3.3 Anti diabetic activity
Inhibition of Alpha-Amylase Enzyme
Diabetes mellitus is a group of metabolic diseases in which there are high blood sugar
levels over a prolonged period. A therapeutic approach to decrease the hyperglycaemia is to
0
10
20
30
40
50
60
70
100 200 300
% o
f in
hib
itio
n
Concentration
Albumen Denaturation
% of Protein Denaturation of the ZnNPs
% of Protein Denaturation of the CuNPs
Aspirin(Standard)
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org640
inhibit the carbohydrate digesting enzymes (α-glucosidase and α-amylase), thereby preventing
the breakdown of carbohydrates into monosaccharides which is a main cause of increasing blood
glucose level. Therefore, developing compounds having inhibitory activities towards
carbohydrate hydrolysing enzymes may be a useful way to manage diabetes. As shown in Figure
4 and Table 5, α-amylase and α-glucosidase were significantly inhibited in a dose-dependent
manner by the ZnNPs and CuNPs. The results suggest that with the increased ZnNPs and CuNPs
concentration, the activity levels of enzyme were remarkably reduced, Hence, the biomolecules
likely enhanced the antidiabetic potential of the synthesized NPs. α-Amylase inhibitory actions
were observed in increasing order, as Acarbose (Figure 5). Comparable results were observed.
However, the foregoing results suggest that the synthesized ZnNPs and CuNPs have potential
antidiabetic property and could prove its effectiveness in the diabetes care.
For the ZnNPs, the result shows the percentage of cytotoxicity for 0.05 mg/ml as 57.03%,
0.1 mg/ml as 60.06% , 0.15 mg/ml as 61.02%, 0.2 mg/ml as 64.02% and 0.25 mg/ml as 60.03%.
For the CuNPs, the result shows the percentage of cytotoxicity for 0.05 mg/ml as 34.02%, 0.1
mg/ml as 43.02% , 0.15 mg/ml as 48.02%, 0.2 mg/ml as 60.02% and 0.25 mg/ml as 61.05%.
These inhibition values are compared with standard drug of Acarbose
for 0.05 mg/ml as 35%, 0.1 mg/ml as 42% , 0.15 mg/ml as 56%, 0.2 mg/ml as 61% and 0.25
mg/ml as 79%.
Albumen Denaturation did not show any significant difference at 0.5 µg/ml and
0.10µg/ml, however, it showed a significant change when the concentration of nanoparticles are
0.15µg/ml, 0.20µg/ml and 0.25 µg/ml concentrations. All the values are compared with standard
drug of Acarbose (Figure 5). On comparing, it was observed that when the concentration of the
sample increases the inhibition also increases showing a good sign of Anti-diabetic activity.
As a part of the investigation on the mechanism of the Anti diabetic activity, ability of
extract to inhibit Inhibition of Alpha-Amylase Enzyme was studied. The in-vitro study of Anti
diabetic activity indicates that the inhibition percentage of Alpha-Amylase Enzyme by Acalypha
indica root extracts of ZnNPs activity is higher than CuNPs.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org641
Table.4 Anti diabetic activity of ZnNPs and CuNPs using root extracts of Acalypha indica
by Alpha amylase.
Fig.5 Graphical representation of Anti diabetic activity of ZnNPs and CuNPs using root
extracts of Acalypha indica by Alpha amylase.
0
10
20
30
40
50
60
70
80
90
0.05 0.1 0.15 0.2 0.25
% o
f in
hib
itio
n
Concentration
Inhibition of Alpha-Amylase Enzyme
% of inhibitionof the ZnNPs
% of inhibitionof the CuNPs
Acarbose(Standard)
S.No Test
Concentration
of the sample
(µg/ml)
% of inhibition
of the ZnNPs
% of
inhibition of
the CuNPs
Acarbose
(Standard)
1
Alpha amylase
inhibitory
activity
0.5 57.03 34.02 35
2 0.1 60.06 43.02 42
3 0.15 61.02 48.02 56
4 0.2 64.02 60.02 61
5 0.25 60.03 61.05 79
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org642
4. Conclusion
Scientists have shifted their interest from chemical or physical methods to biological methods as
it does not involve a combination of abusive or toxic chemicals to human health or any
involvement of immense machines or equipment. The biological methods incorporate other plant
or microbial mediated methods that are cheap and easily accessible in daily life. The medicinal
plant Acalypha indica has been used as a traditional medicinal plant due to the presence of
phytochemicals in it. The various applications of the root extract have already been established
till date. Now, in this study, the root extracts have been used for the biogenesis of the ZnNPs and
CuNPs nanoparticles. The DPPH assay is the most acceptable, fastest and simplest method for
the calculation of the free radical scavenging activity. As shown in the Table 2 and Figure 3. the
ZnNps and CuNps shows better antioxidant property when compared with the standard ascorbic
acid with an IC50 values. Denaturation of proteins is a well-documented cause of inflammation.
Phenylbutazones, salicylic acid, flufenamic acid (anti‐inflammatorydrugs), have shown dose
dependent ability to thermally induced protein denaturation. As a part of the investigation on the
mechanism of the anti‐inflammatory activity, ability of extract to inhibit protein denaturation
was studied. The in-vitro study of a anti-inflammatory activity indicates that the inhibition
percentage of albumin denaturation by Acalypha indica Root extracts. It is inferred that the
Anti- inflammatory activity of CuNPs synthesized from Acalypha indica root extracts indicates
a good and higher inhibition percentage than ZnNPs from Acalypha indica root extracts as
presented in Table 3 and Figure 4. α-amylase is a key enzyme in carbohydrate metabolism.
Inhibition of α-amylase is one of the strategies for treating diabetes. Amylase inhibitors are also
known as starch blockers because they contain substances that prevent dietary starches from
being absorbed by the body. Amylase inhibitor with starchy meal will reduce the usual rise in
blood sugar levels. The result suggests that Zn Nanoparticle exhibits well α- amylase inhibition
under in vitro conditions (Tables 4 and Fig 5).
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org643
5. References
1. Pallab Maity, Dhananjay Hansda, Uday Bandyopadhyay, Dipak Kumar Mishra, Indian
Journal of Experimental Biology, 2009, 47, 849-861.
2. R. K. Gupta, Medicinal & Aromatic plants, CBS publishers & distributors, 1st edition,
2010, 116-117.
3. Ahmad A., et al. “Extracellular biosynthesis of silver nanoparticles using the fungus
Fusarium oxysporum”. Colloids and Surfaces B: Biointerfaces, 2003, 28.4, 313-318.
4. Padalia H and Chanda S. “Green synthesis of silver nanoparticles from marigold flower
and its synergistic antimicrobial potential”. Arabian Journal of Chemistry, 2015, 8.5,
732-741.
5. Krishnaraj C., et al. “Synthesis of silver nanoparticles using Acalypha indica leaf
extracts and its antibacterial activity against water borne pathogens”. Colloids and
Surfaces B: Biointerfaces,2010, 76.1, 50-56.
6. Christensen L, Vivekanandhan S, Misra M, Mohanty AK. Biosynthesis of silver
nanoparticles using Murraya koenigii (curry leaf): an investigation on the effect of broth
concentration in reduction mechanism and particle size. Adv Mater Lett 2011,2,429.
7. 10. Ali ZA, Yahya R, Sekaran SD, Puteh R. Green synthesis of silver nanoparticles using
apple extract and its antibacterial properties. Adv Mater Sci Eng 2016,6.
8. 11. Rajaselvam J, Benila Smily JM, Meena R. A study of antimicrobial activity of
Acalypha indica against selected microbial species. Int J Life Sci Pharma Res 2012,
3,473-6.
9. Varier, V.P.S. Indian medicinal plants: a compendium of 500 species Orient Longman
Publication, Madras, India, 1996, p. 134.
10. Perry, L.M. Medicinal plants of East and Southeast Asia: attributed properties and uses.
MIT Press, Cambridge. Mass. U.S.A., 1980, p. 109.
11. Winter, H., Griffith, M. D. Vitamins, Herbs, Minerals, and Supplements: The Complete
Guide. Fisher Books. USA, 1998, p. 217.
12. Kirtikar, K.R., Basu, B.D. Indian Medical Plants. Volume II. Second Edition. Jayyed
Press, New Delhi, 1975, p. 30-45.
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org644
13. Ramachandran, J. Herbs of Siddha Medicine/The First 3D Book On Herbs. Murugan
PPatthipagam, Chennai, India, 2008, p. 156.
14. Rahman, M.A., Bachar, S.C. and M. Rahmatullah. Analgesic and antiinflammatory
activity of methanolic extract of Acalypha indica Linn. Pak. J. Pharm. Sci., 2010, 23(3),
256-258.
15. Govindarajan, M., Jebanesan, A., Reetha, D., Amsath, R., Pushpanathan, T., and K.
Samidurai, Antibacterial activity of Acalypha indica L. Eur. Rev. Med. Pharmacol. Sci.,
2008, 12(5), 299-302.
16. Krishnaraj, C., Jagan, E.G., Rajasekar, S., Selvakumar, P., Kalaichelvan, P.T., Mohan
N., Synthesis of silver nanoparticles using Acalypha indica leaf extracts and its
antibacterial activity against water borne pathogens. Colloids and Surfaces B:
Biointerfaces, 2010, 76(1),50-56.
17. Krishnaraj C, Muthukumaran P, Ramachandran R, Balakumaran MD, Kalaichelvan PT
Acalypha indica Linn: Biogenic synthesis of silver and gold nanoparticles and their
cytotoxic effects against MDA-MB-231, human breast cancer cells. Biotechnol Reports
2014,4, 42-49.
18. T.J. Beveridge, M.N. Hughes, H. Lee, K.T. Leung, R.K. Poole, I. Savvaidis, S. Silver, J.. T
revorsMetal-microbe interactions: contemporary approaches Adv. Microb. Physiol.,
1997, 38 p. 178
19. M.I. Sriram, K. Kalishwaralal, S. GurunathanBiosynthesis of silver and gold
nanoparticles using Bacillus licheniformis Method Mol. Biol., 2012, 906, p. 33-43
20. G. Li, D. He, Y. Qian, B. Guan, CuiY. Gao, K. Yokoyama, L. WangFungus-mediated
green synthesis of silver nanoparticles using Aspergillus terreus Int. J. Mol. Sci.,
2012, 13, p. 466-476
21. Chauhan, S. Zubair, S. Tufail, A. Sherwani, M. Sajid, S.C. Raman, A. Azam, M. OwaisFu
ngus-mediated biological synthesis of gold nanoparticles: potential in detection of liver
cancer Int. J. Nanomedicine, 2011, 6, p. 2305-2319
22. T. Elavazhagan, K.D. ArunachalamMemecylon edule leaf extract mediated green
synthesis of silver and gold nanoparticles Int. J. Nanomedicine, 2011, 6, p. 1265-1278
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org645
23. D.M. Ali, N. Thajuddin, K. Jeganathan, M. GunasekaranPlant extract mediated synthesis
of silver and gold nanoparticles and its antibacterial activity against clinically isolated
pathogens Colloids Surf. B, 2011, 85, p. 360-365
24. V. Kumar, S.K. YadavPlant-mediated synthesis of silver and gold nanoparticles and their
applications J. Chem. Technol. Biotechnol., 2009, 84, p. 151-157
25. .B. Narayanan, N. SakthivelCoriander leaf mediated biosynthesis of gold nanoparticles
Mater. Lett., 2008, 62, p.4588-4590
26. R. Kalaiarasi, N. Jayalakshmi, P. Venkatachalam Phytosynthesis of nanoparticles and its
applications Plant Cell Biotechnol. Mol. Biol., 2010, 11, p. 1-16
Journal of Information and Computational Science
Volume 10 Issue 4 - 2020
ISSN: 1548-7741
www.joics.org646