Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (5): 546-554.doi: 10.3736/jcim20120510

• Original Experimental Research • Previous Articles     Next Articles

Rapid green synthesis of silver nanoparticles from silver nitrate by a homeopathic mother tincture Phytolacca Decandra

Soumya Sundar Bhattacharyya,Jayeeta Das,Sreemanti Das,Asmita Samadder,Durba Das,Arnab De,Saili Paul,Anisur Rahman Khuda-Bukhsh#br#   

  1. Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, IndiaCytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India
  • Received:2011-12-05 Accepted:2012-01-30 Online:2012-05-20 Published:2018-06-15

Objective: To examine if a homeopathic mother tincture (Phytolacca Decandra) is capable of precipitating silver nanoparticles from silver nitrate (AgNO3) and to characterize the biosynthesized nanoparticles for evaluating their biological activities.

Methods: A total of 100 mg of AgNO3 was added to 20 mL of Milli-Q water and stirred vigorously. Then 5 mL of the homeopathic mother tincture of Phytolacca Decandra (ethanolic root extract of Phytolacca decandra) was added and stirred continuously. Reduction took place rapidly at 300 K and completed in 10 min as shown by stable light greenish-yellow color of the solution which gave colloid of silver nanoparticles. The colloid solution was then centrifuged at 5 000×g to separate the nanoparticles for further use. The nanoparticles were characterized by spectroscopic analysis, particle size analysis and zeta potential measurements, and morphology was analyzed by atomic force microscopy. The drug-DNA interaction was determined by circular dichroism spectrophotometry and melting temperature profiles by using calf thymus DNA as the target. The biological activities were determined using a cancer cell line A549 in vitro and using bacteria Escherichia coli and fungus Saccharomyces cerevisiae as test models.

Results: Phytolacca Decandra precipitated silver nanoparticles in ambient conditions. The nanoparticles had 91 nm particle size, with polydispersity index of 0.119 and zeta potential of –15.6 mV. The silver nanoparticles showed anticancer and antibacterial properties, but no clear antifungal properties.

Conclusion: This could be a novel environment-friendly method to biosynthesize silver nanoparticles using a cost-effective, nontoxic manner. The homeopathic mother tincture may utilize this property of nano-precipitation in curing diseases or disease symptoms.

Key words: silver nitrate, metal nanoparticles, Phytolacca decandra, homeopathy, biosynthetic pathways


Name of primers Sequence of primers
Caspase 3 Forward: 5′-AGGGGTCATTTATGGGACA-3′

Figure 1

Surface images by atomic force microscopy with average particle size by DLS analysis of SPs A: 2D image of SPs; B: 3D image of SPs; C: Average particle size of SPs obtained from DLS data; D: Positive and negative control experimental setup to prepare nanoparticles from silver nitrate (a: Normal silver nitrate without any treatment; b: Silver nitrate plus placebo (45% ethanol; negative control); c: Silver nitrate plus Phytolacca Decandra; d: Silver nitrate plus ascorbic acid (positive control)). SP: silver nanoparticle; DLS: dynamic light scattering."

Figure 2

Ultraviolet-visible spectral image of silver nanoparticles"

Figure 3

Circular dichroism spectrum of calf thymus DNA incubated with or without silver nanoparticles"

Figure 4

Melting curves of calf thymus DNA in presence and absence of silver nanoparticlesCT: calf thymus"

Figure 5

Cell viability of A549 cells with different concentrations of SPs Data are presented as mean±standard deviation, n=3; *P<0.05, vs control group. SP: silver nanoparticle."

Figure 6

Surface morphology of A549 cells (Scanning electron microscopy, ×3 500) C: A549 cells without any treatment; SP1: 80 μg/mL of SPs; SP2: 100 μg/mL of SPs. SP: silver nanoparticle."

Figure 7

PI staining (A) and DAPI staining (B) of A549 cells (Fluorescence microscopy, ×200) C: A549 cells without any treatment; SP1: 80 μg/mL of SPs; SP2: 100 μg/mL of SPs. SP: silver nanoparticle; PI: propidium iodide; DAPI: 4′,6-diamidino-2-phenylindole."

Figure 8

DNA damage tested by comet assay (Fluorescence microscopy, ×400) C: A549 cells without any treatment; SP1: 80 μg/mL of SPs; SP2: 100 μg/mL of SPs. SP: silver nanoparticle."

Figure 9

DNA fragmentation tested by agarose gel electrophoresis Ln1: A549 cells without any treatment; Ln2: 80 μg/mL of SPs; Ln3: 100 μg/mL of SPs. SP: silver nanoparticle."

Figure 10

RT-PCR analysis of G3PDH and caspase 3 Ln1: A549 cells without any treatment; Ln2: 80 μg/mL of SPs; Ln3: 100 μg/mL of SPs. SP: silver nanoparticle. RT-PCR: reverse transcription-polymerase chain reaction; G3PDH: glyceraldehyde-3-phosphate dehydrogenase."


Concentration of SPs (mg/L) Antibacterial effects of SPs Antifungal effects of SPs
20 + +
40 + +
80 +
120 +
160 +
200 +
400 +
[1] Huang J, Li Q, Sun D, Lu Y, Su Y, Yang X, Wang H, Wang Y, Shao W, He N, Hong J, Chen C . Biosynjournal of silver and gold nanoparticles by novel sundried Cinnamomum camphora leaf[J]. Nanotechnology, 2007,18(10):105104.
doi: 10.1088/0957-4484/18/10/105104
[2] Vaidyanathan R, Kalishwaralal K, Gopalram S, Gurunathan S . Nanosilver ― the burgeoning therapeutic molecule and its green synjournal[J]. Biotechnol Adv, 2009,27(6):924-937.
doi: 10.1016/j.biotechadv.2009.08.001 pmid: 19686832
[3] Jha AK, Prasad K, Prasad K, Kulkarni AR . Plant system: nature’s nanofactory[J]. Colloid Surf B Biointerfaces, 2009,73(2):219-223.
doi: 10.1016/j.colsurfb.2009.05.018 pmid: 19539452
[4] Gardea-Torresdey JL, Parsons JG, Gomez E, Peralta-Videa J, Troiani HE, Santiago P, Jose Yacaman M . Formation and growth of Au nanoparticles inside live alfalfa plants[J]. Nano Lett, 2002,2(4):397-401.
doi: 10.1021/nl015673+
[5] Mohanpuria P, Rana NK, Yadav SK . Biosynjournal of nanoparticles: technological concepts and future applications[J]. J Nanopart Res, 2008,10(3):507-517.
doi: 10.1007/s11051-007-9275-x
[6] Venugopal Rao S . Picosecond nonlinear optical studies of gold nanoparticles synjournaled using coriander leaves(Coriandrum sativum)[J]. J Mod Opt, 2011,58(12):1024-1029.
doi: 10.1080/09500340.2011.590903
[7] Kasthuri J, Veerapandian S, Rajendiran N . Biological synjournal of silver and gold nanoparticles using apiin as reducing agent[J]. Colloids Surf B Biointerfaces, 2009,68(1):55-60.
doi: 10.1016/j.colsurfb.2008.09.021 pmid: 18977643
[8] Sathishkumar M, Sneha K, Won SW, Cho CW, Kim S, Yun YS . Cinnamon zeylanicum bark extract and powder mediated green synjournal of nano-crystalline silver particles and its bactericidal activity[J]. Colloids Surf B Biointerfaces, 2009,73(2):332-338.
doi: 10.1016/j.colsurfb.2009.06.005
[9] Philip D . Biosynjournal of Au, Ag and Au-Ag nanoparticles using edible mushroom extract[J]. Spectrochim Acta A Mol Biomol Spectrosc, 2009,73(2):374-381.
doi: 10.1016/j.saa.2009.02.037 pmid: 19324587
[10] European Pharmacopoeia Commission . The European Pharmacopoeia: elaborations and revisions. Guide for the work of the European Pharmacopoeia. (PA/PH/ SG (10) 42 DEF). 2010.
[11] Bhattacharyya SS, Paul S, De A, Das D, Samadder A, Boujedaini N, Khuda-Bukhsh AR . Poly(lactide-co-glycolide) acid nanoencapsulation of a synthetic coumarin: cytotoxicity and bio-distribution in mice, in cancer cell line and interaction with calf thymus DNA as target[J]. Toxicol Appl Pharmacol, 2011,253(3):270-281.
doi: 10.1016/j.taap.2011.04.010
[12] Mossann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays[J]. J Immunol Methods, 1983,65(1-2):55-63.
doi: 10.1016/0022-1759(83)90303-4
[13] Biswas R, Mandal SK, Dutta S, Bhattacharyya SS, Boujedaini N, Khuda-Bukhsh AR . Thujone-rich fraction of Thuja occidentalis demonstrates major anti-cancer potentials: evidences from in vitro studies on A375 cells. Evid Based Complement Alternat Med. 2011; 568148.
[14] Lee HJ, Yeo SY, Jeong SH . Antibacterial effect of nanosized silver colloidal solution on textile fabrics[J]. J Mater Sci, 2003,38(10):2199-2204.
doi: 10.1023/A:1023736416361
[15] Lanone S, Boczkowski J . Biomedical applications and potential health risks of nanomaterials: molecular mechanisms[J]. Curr Mol Med, 2006,6(6):651-663.
doi: 10.2174/156652406778195026
[16] Atiyeh BS, Costagliola M, Hayek SN, Dibo SA . Effect of silver on burn wound infection control and healing: review of the literature[J]. Burns, 2007,33(2):139-148.
doi: 10.1016/j.burns.2006.06.010
[17] Anil Kumar S, Abyaneh MK, Gosavi SW, Kulkarni SK, Pasricha R, Ahmad A, Khan MI . Nitrate reductase-mediated synjournal of silver nanoparticles from AgNO3[J]. Biotechnol Lett, 2007,29(3):439-445.
doi: 10.1007/s10529-006-9256-7 pmid: 17237973
[18] Law N, Ansari S, Livens FR, Renshaw JC, Lloyd JR . Formation of nanoscale elemental silver particles via enzymatic reduction by Geobacter sulfurreducens[J]. Appl Environ Microbiol, 2008,74(22):7090-7093.
doi: 10.1128/AEM.01069-08
[19] Kauffman CA, Carver PL . Antifungal agents in the 1990s. Current status and future developments[J]. Drugs, 1997,53(4):539-549.
doi: 10.2165/00003495-199753040-00001
[20] Mackay CR . Moving targets: cell migration inhibitors as new anti-inflammatory therapies[J]. Nat Immunol, 2008,9(9):988-998.
doi: 10.1038/ni.f.210 pmid: 18711436
[21] Lu L, Sun RW, Chen R, Hui CK, Ho CM, Luk JM, Lau GK, Che CM . Silver nanoparticles inhibit hepatitis B virus replication[J]. Antivir Ther, 2008,13(2):253-262.
doi: 10.1016/j.antiviral.2007.08.009 pmid: 18505176
[22] Peer D, Karp JM, Hong S, Farokhzad OC, Margalit R, Langer R . Nanocarriers as an emerging platform for cancer therapy[J]. Nat Nanotechnol, 2007,2(12):751-760.
doi: 10.1038/nnano.2007.387 pmid: 18654426
[1] Ubiratan Cardinalli Adler, Maristela Schiabel Adler, Ana Elisa Madureira Padula, Livia Mitchiguian Hotta, Amarilysde Toledo Cesar, José Nelson Martins Diniz, Helen de Freitas Santos, Edson Zangiacomi Martinez. Homeopathy for COVID-19 in primary care: A randomized, double-blind, placebo-controlled trial (COVID-Simile study). Journal of Integrative Medicine, 2022, 20(3): 221-229.
[2] Mikhail Teppone. Medicine has always been “Modern” and “Scientific” from ancient times to the present day. Journal of Integrative Medicine, 2019, 17(4): 229-237.
[3] Ubiratan Cardinalli Adler, Ana Carolina Acorinte, Fernando Oliveira Calzavara, Adriano Andréda Silva, Amarilys de Toledo Cesar, Maristela Schiabel Adler, Edson Zangiacomi Martinez, José Carlos Fernandes Galduróz. Double-blind evaluation of homeopathy on cocaine craving: A randomized controlled pilot study. Journal of Integrative Medicine, 2018, 16(3): 178-184.
[4] Shubha Singhal, Vandana Roy. Awareness, practice and views about integrating AYUSH in allopathic curriculum of allopathic doctors and interns in a tertiary care teaching hospital in New Delhi, India. Journal of Integrative Medicine, 2018, 16(2): 113-119.
[5] Salvatore Chirumbolo, Geir Bjørklund. Homeopathic potencies of Arnica montana L. change gene expression in a Tamm-Horsfall protein-1 cell line in vitro model: The role of ethanol as a possible confounder and statistical bias. Journal of Integrative Medicine, 2017, 15(4): 255-264.
[6] Saeed Ahmad, Tayyeba Rehman, Waheed Mumtaz Abbasi. Effects of homoeopathic ultrahigh dilutions of Aconitum napellus on Baker's yeast-induced fever in rabbits. Journal of Integrative Medicine, 2017, 15(3): 209-213.
[7] Jesmin Mondal, Asmita Samadder, Anisur Rahman Khuda-Bukhsh. Psorinum 6× triggers apoptosis signals in human lung cancer cells. Journal of Integrative Medicine, 2016, 14(2): 143-153.
[8] Santu Kumar Saha, Sourav Roy, Anisur Rahman Khuda-Bukhsh. Ultra-highly diluted plant extracts of Hydrastis canadensis and Marsdenia condurango induce epigenetic modifications and alter gene expression profiles in HeLa cells in vitro. Journal of Integrative Medicine, 2015, 13(6): 400-411.
[9] Subhranil Saha​, Munmun Koley​, Subhasish Ganguly​, Prasanta Rath​, Pulak Roy Chowdhury, Seikh Intaj Hossain. Developing the criteria for evaluating quality of individualization in homeopathic clinical trialreporting: A preliminary study. Journal of Integrative Medicine, 2014, 12(1): 13-19.
[10] Subhranil Saha​, Munmun Koley, Jogendra Singh Arya, Gurudev Choubey, Shubhamoy Ghosh​, Subhasish Ganguly​, Aloke Ghosh​, Sangita Saha​, Malay Mundle​. Medicine prescription practices of homeopathic undergraduate students in West Bengal, India. Journal of Integrative Medicine, 2014, 12(1): 7-12.
[11] Munmun Koley, Subhranil Saha, Jogendra Singh Arya, Gurudev Choubey, Shubhamoy Ghosh, Rajib Purkait, Ramkumar Mondal, Bapi Kundu, Rajarshi Mukherjee. A study on drug utilization and prescription habits of physicians in a government homeopathic hospital in West Bengal, India. Journal of Chinese Integrative Medicine, 2013, 11(5): 305-313.
[12] Samrat Ghosh, Kausik Bishayee, Avijit Paul, Avinaba Mukherjee, Sourav Sikdar, Debrup Chakraborty, Naoual Boujedaini, Anisur Rahman Khuda-Bukhsh. Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation. Journal of Chinese Integrative Medicine, 2013, 11(2): 116-124.
[13] Yogesh Bendale, Vineeta Bendale, Saili Paul, Soumya Sundar Bhattacharyya. Green synthesis, characterization and anticancer potential of platinum nanoparticles Bioplatin. Journal of Chinese Integrative Medicine, 2012, 10(6): 681-689.
[14] Santu Kumar Saha, Sreemanti Das, Anisur Rahman Khuda-Bukhsh. Phenotypic evidence of ultra-highly diluted homeopathic remedies acting at gene expression level: A novel probe on experimental phage infectivity in bacteria. Journal of Chinese Integrative Medicine, 2012, 10(4): 462-470.
[15] Das Sreemanti, Kumar Saha Santu, De Arnab, Das Durba, Rahman Khuda-Bukhsh Anisur. Potential of the homeopathic remedy, Arnica Montana 30C, to reduce DNA damage in Escherichia coli exposed to ultraviolet irradiation through up-regulation of nucleotide excision repair genes. Journal of Chinese Integrative Medicine, 2012, 10(3): 337-246.
Full text



[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239