EVALUATION OF HEAVY METAL DISTRIBUTION AND ANTIBACTERIAL ACTIVITIES OF MEDICINAL PLANTS TINOSPORA CORDIFOLIA, OCIMUM SANCTUM AND PIPER NIGRUM
Abstract
To evaluate the inhibition and the in vitro interaction of the pathogens with the plant extracts, various solvent extracts of Tinospora cordifolia, Ocimum sanctum, Piper nigrum were screened for antimicrobial activity using agar well diffusion method. A synergistic inhibitory activity by the combination of aqueous extracts of the three plants against Salmonella, Shigella and P. aeruginosa was observed. The chloroform extracts in combination also showed maximum inhibition against E. coli but ethanolic extract in equimolar concentration showed maximum inhibition against Staphylococcus aureus. All these plant extracts were tested for heavy metal (HM) content as they are often contaminated during the growth, development and processing. O. sanctum and P. nigrum contained the maximum HM content while T. cordifolia contained less HM. The plants extracts in combination also showed low contamination with heavy metal which is a positive indication for their potential use in combating infections. These results indicate that plant extracts in combinations are less contaminated with heavy metals but they are very potent in fighting emerging pathogens.
Keywords:
Antimicrobial, heavy metal, Tinospora cordifolia, Ocimum sanctum, Piper nigrum, Minimum inhibitory concentration, plant extractDOI
https://doi.org/10.25004/IJPSDR.2014.060312References
1. Borde VU, Pawar DP, Shelar SR, Apturkar RM. Antimicrobial activity of some medicinal plants. Sci. Res. Rep. 2013; 3(1):33-37.
2. Issazedeh K, Massiha A, Pahlaviani MRMK. Minimum inhibitory concentration (MIC) of Myrtus communis extract and nystatin on clinical isolated and standard strains of Candida albicans. J. Appl. Environ. Biol. Sci.2012; 2(9):466-468.
3. Biswas P. evaluation of antibacterial activities of leaf extract of two medicinal plants Ocimum canum sims and Ocimum tenuiflorum Linn. J. Microbiol Biotech Res. 2013; 3(3):20-23.
4. Chandra R, Dwivedi V, Shivam K, Jha AK. Detection of antimicrobial activity of Ocimum sanctum (Tulsi) and Trigonella foenum graecum (Methi) against selected bacterial and fungal strains. Research J Pharma., Bio.& Chem. Sci. 2011; 2(4): 809-813.
5. Debnath M. Clonal propagation and antimicrobial activity of an endemic medicinal plant Stevia rebaudiana. J Medl Plts Res 2008; 2(2): 45-51.
6. Mohan L, Amberkar MV, Kumari M. Ocimum sanctum linn (Tulsi)- an overview. Int. J. Pharma. Sci. Rev. & Res. 2011; 7(1): 51-53.
7. Rajeshwari S. Ocimum sanctum , the Indian home remedy, In Current medicinal scnene, Rajeshwari Foundation Limited Bombay. 1992.
8. Hussain AI, Anwar F, Sherazi STH , Przybylski R. Chemical composition. Antioxidant and antimicrobial activities of basil (Ocimum basilicum) essential oils depends on seasonal variations. Food Chem. 2008; 108: 986-995.
9. Chattopadhyay RR, Sarkar SK, Ganguly S, Medda C, Basu TK. Hepatoprotective activity of Ocimum sanctum leaf extract against paracetamol induced hepatic damage in rats. Ind. J. Pharmacol. 1992; 24: 163-165.
10. Singh AR, Bajaj VK, Shekhawat VS, Singh K. Phytochemical estimation and Antimicrobial activity of Aqueous and Methanolic extract of Ocimum sanctum L. J. Nat. Prod. Plant Resour. 2013; 3 (1):51-58.
11. Ali H, Dixit S. In-vitro antimicrobial activity of flavanoids of Ocimum sanctum with synergistic effect of their combined form. Asian Pacific J of Trop. Dis. 2012; S396-S398.
12. Agarwal P, Murlikrishnan NL. Evaluation of the antimicrobial activity of various concentrations of Tulsi (Ocimum sanctum) extract against Streptococcus mutans: an in-vitro study. Indian J Dent Res. 2010; 21(3):357-9.
13. Mishra P, Mishra S. Study of Antibacterial Activity of Ocimum sanctum Extract Against Gram Positive and Gram Negative Bacteria. Amer J of Food Tech, 2011; 6: 336-341.
14. Sindhu S, Manorama S, Alfamol PM. Preliminary phytochemical analysis and antimicrobial activity of Piper longum L. (Piperaceae) Mintage. J of Pharma & Med Sci. 2013; 2(1): 21-23.
15. Karsha PV, Lakshmi OB. Antibacterial activity of black pepper (Piper longum Linn.) with special reference to its mode of action on bacteria. Ind J of Nat Prod & Res. 2010;1(2): 213-215.
16. Williamson EM. Major herbs of Ayurveda Churchill living stone, Elsevier Science Ltd, London, 2002.
17. Joy B, Sandhya CP, Remitha KR. Comparison and bioevaluation of Piper longum fruit extracts. J. Chem. Pharm. Res. 2010; 2(4):696-706.
18. Singh C, Rai NP. In-vitro Antibacterial Activity of Piper longum L. Fruit. Int. J. Pharm. Sci. Rev. Res. 2013; 18(2): 89-91.
19. Khan M, Siddique. Antimicrobial activity of piper fruits. Natural products radiance 2007; 6(2): 111-113.
20. Molla AH, Zakaria MG , Molla MTH, Alam MT, Ahsan MS. Chemical investigations and microbial activity of Tinospora cordifolia miers. J. Bio. Sci. 2012; 29:153-160.
21. Singh SS, Pandey SC, Srivastava S, Gupta VS, Patro B, Ghosh AC. Chemistry and medicinal properties of Tinospora cordifolia (Guduchi). Indian J. Pharmacol. 2003; 35: 83-91.
22. Alam P, Ali M, Singh R, Madhurima, Ahmad S, Shakeel F. A validated HPLC method for estimation of cordifolioside A in Tinospora cordifolia, Miers and marketed formulations. J Chromatogr Sci. 2009; 47(10):910-3.
23. Jeyachandran R, Xavier TF, Anand SF. Antibacterial activity of stem extracts of Tinospora cordifolia (Willd) Hook. f & Thomson. Anc Sci Life. 2003; 23(1): 40-43.
24. Gahlawat D, Mazumdar A. In-vitro antibacterial activity of the whole plant of Tinospora cordifolia. Int J of Health and Pharma Sci.2013; 2(3): 1-6.
25. Upadhay RK, Tripathi R, Ahmad S. Antimicrobial activity of two Indian medicinal plants Tinospora cordifolia (family Menispermaceae) and Cassia fistula (Family: Caesalpinaceae) against human pathogenic bacteria. J of Phar Res. 2011; 4(1):167-170.
26. Lekouch N, Sedki A, Nejmeddine A, Gamon S. Lead and traditional Moroccan pharmacopoeia. Sci Total Environ. 2001; 280:39-43.
27. Dwivedi SK, Dey S. Medicinal herbs: A potential source of toxic metal exposure for man and animals in India. Arch Environ Health. 2002; 57:229-31.
28. Gajalakshmi S, Iswarya V, Ashwini R, Divya G, Mythili S, Sathiavelu A. Evaluation of heavy metals in medicinal plants growing in Vellore District. Eur J of Exp Biol. 2012; 2(5):1457-1461.
29. Dzomba P, Chayamiti T, Togarepi E. Heavy Metal Content of Selected Raw Medicinal Plant Materials: Implication for Patient Health. Bull. Environ. Pharmacol. Life Sci. 2012; 1(10): 28-33.
30. Sivananthan M .Antibacterial activity of 50 medicinal plants used in folk medicine International Journal of Biosciences 2013; 3(4):104-121.
31. IS 5887 (Part 1). Isolation, Identification and Enumeration of E. coli.1976 (Reaffirmed 2005).
32. IS 14843. Meat and meat products – Enumeration of Pseudomonas spp. 2000.
33. IS 5887 (Part 3). General Guidance on Methods for the Detection of Salmonella. 1999 (Reaffirmed 2005).
34. IS 5887 (Part 7).General Guidance on Methods for the Isolation and Identification of Shigella. 1999 (Reaffirmed 2005).
35. IS 5887(Part 2). Isolation, Identification and Enumeration of S. aureus and F. streptococci. 1976 (Reaffirmed 2005).
36. Debnath M, Chejara V, Vijaya BK, Bika D, Sajal K, Dixit A, Jain R, Jain V. Evaluation of quality and impact of untreated wastewater for irrigation. Amer J Res Com. 2014: 2(4): 200-231.
37. Temminghoff EJM, Houba VJG. Plant analysis procedures. 2nd ed. Dordrecht: Kluwer Academic Publishers. 2004.
38. Joshi B, Lekhak S, Sharma A. Antibacterial Property of Different Medicinal Plants: Ocimum sanctum, Cinnamomum zeylanicum, Xanthoxylum armatum and Origanum majorana. Kat Uni J of Sci Eng and Tech. 2009; 5(1): 143- 150.
39. Bhatt MK, Shankar MB, Saluja AK, Dholwani KK, Captain AD. Evaluation of antimicrobial activity of Ocimum sanctum methanolic extract. J Pharm Sci Innov. 2012; 1(4): 39-41.
40. Abeysinghe PD, Wanigatunge RP. Evaluation of antibacterial activity of different mangrove plant extracts. Ruh J of Sci. 2006; 1:104-112.
41. Kamegam N, Karuppusamy S, Prakash M, Jayakumar M, Rajasekar K. Antibacterial potency and synergistic effects of certain plant extracts against food borne diarrheagenic bacteria. Int J of biomed and pharma sci. 2008:2(2): 88-93.
42. Rivera SEV, Escobar-Saucedo MA, Morales D, Aguilar CN, Herrera RR. Synergistic effects of ethanolic plant extract mixtures against food-borne pathogen bacteria. Afr J of Biotech.2014; 13(5):699-704.
43. Sahu M, Sujatha S, Chaya DR, Parija SC. Pericardial effusion–an unusual manifestation of salmonellosis: a case report. Cases J. 2008; 1:375.
44. Fontaine A, Arondel J, Sansonetti PJ. Construction and evaluation of live attenuated vaccine strains of Shigella flexneri and Shigella dysenteriae. Res Microbiol.1990; 141(7-8):907-12.
45. Lunavath V, Porika R, Priya S , Mamidala E Preliminary phytochemical analysis and anti bacterial activity of Tinospora cordifolia leaf extracts. Inter J of Health and Pharma Sci. 2012; 1(4):18-21.
46. Zhang W, Robertson DC, Zhang C, Bai W, Zhao M, Francis DH. Escherichia coli constructs expressing human or porcine enterotoxins induce identical diarrheal diseases in a piglet infection model. Appl. Environ. Microbiol.2008; 74:5832-5837.
47. Ahn SH, Tsalik EL, Cyr DD, Zhang Y, van Velkinburgh JC, Langley RJ, Glickman SW, Cairns CB, Zaas AK, Rivers EP, Otero RM, Veldman T, Kingsmore SF, Lucas J, Woods CW, Ginsburg GS, Fowler VG Jr. Gene expression-based classifiers identify Staphylococcus aureus infection in mice and humans. PLoS One. 2013; 8(1):e48979.
48. Shamina A, Sarma YR. Secondary metabolites in black pepper (Piper nigrum) and their effect on the foot-rot pathogen Phytophthora capsici. J of Plantation Crops, 2001; 29 (2): 22-26.
49. Singh V, Amdekar S, Verma O. Ocimum sanctum (tulsi): Bio-pharmacological Activities. Pharmacol. 2010; 1(10):WMC001046.
50. Mishra A, Kumar S, Pandey AK. Scientific Validation of the Medicinal Efficacy of Tinospora cordifolia. Scientific World J. 2013(2013); Article 292934.8 pages.
51. WHO: Geneva Switzerland; 1998. Quality control methods for medicinal plant materials. available at http://whqlibdoc.who.int/publications/1998/9241545100.pdf .
52. Nayak P. Aluminum: impacts and disease. Environ Res. 2002; 89(2):101-15.
53. Abdurrahman FI, Tijjani MA, Osuji UO. Proximate content and chemical composition of Ocimum viridis leaf and Ocimum gratissum leaf. Internat Res J of Phar 2012; 3(4): 153-156.
54. Ling LJ, Clark RF, Erickson TB, Trestrail JH. philadelphia: Hanley and Belfus Inc; 2001. Toxicology secrets, 2001, pp. 153-69.
55. Khan MA, Ahmad I, Rahman I. Effect of environmental pollution on heavy metals content of Withania somnifera. J of the Chinese Chem Soc. 2007; 54:339-43.
57. Annan K, Dickson, Amponsah IK, Nooni IK. The heavy metal contents of selected plants sampled from different locations. Pharmacog Res. 2013; 5(2): 103-108
58. Choy CMY, Lam CW, Cheung LT, Briton-Jones CM, Cheung LP, Haines CJ. Infertility, blood mercury concentrations and dietary seafood consumption: A case-control study. BJOG. 2002;109:1121-5.
2. Issazedeh K, Massiha A, Pahlaviani MRMK. Minimum inhibitory concentration (MIC) of Myrtus communis extract and nystatin on clinical isolated and standard strains of Candida albicans. J. Appl. Environ. Biol. Sci.2012; 2(9):466-468.
3. Biswas P. evaluation of antibacterial activities of leaf extract of two medicinal plants Ocimum canum sims and Ocimum tenuiflorum Linn. J. Microbiol Biotech Res. 2013; 3(3):20-23.
4. Chandra R, Dwivedi V, Shivam K, Jha AK. Detection of antimicrobial activity of Ocimum sanctum (Tulsi) and Trigonella foenum graecum (Methi) against selected bacterial and fungal strains. Research J Pharma., Bio.& Chem. Sci. 2011; 2(4): 809-813.
5. Debnath M. Clonal propagation and antimicrobial activity of an endemic medicinal plant Stevia rebaudiana. J Medl Plts Res 2008; 2(2): 45-51.
6. Mohan L, Amberkar MV, Kumari M. Ocimum sanctum linn (Tulsi)- an overview. Int. J. Pharma. Sci. Rev. & Res. 2011; 7(1): 51-53.
7. Rajeshwari S. Ocimum sanctum , the Indian home remedy, In Current medicinal scnene, Rajeshwari Foundation Limited Bombay. 1992.
8. Hussain AI, Anwar F, Sherazi STH , Przybylski R. Chemical composition. Antioxidant and antimicrobial activities of basil (Ocimum basilicum) essential oils depends on seasonal variations. Food Chem. 2008; 108: 986-995.
9. Chattopadhyay RR, Sarkar SK, Ganguly S, Medda C, Basu TK. Hepatoprotective activity of Ocimum sanctum leaf extract against paracetamol induced hepatic damage in rats. Ind. J. Pharmacol. 1992; 24: 163-165.
10. Singh AR, Bajaj VK, Shekhawat VS, Singh K. Phytochemical estimation and Antimicrobial activity of Aqueous and Methanolic extract of Ocimum sanctum L. J. Nat. Prod. Plant Resour. 2013; 3 (1):51-58.
11. Ali H, Dixit S. In-vitro antimicrobial activity of flavanoids of Ocimum sanctum with synergistic effect of their combined form. Asian Pacific J of Trop. Dis. 2012; S396-S398.
12. Agarwal P, Murlikrishnan NL. Evaluation of the antimicrobial activity of various concentrations of Tulsi (Ocimum sanctum) extract against Streptococcus mutans: an in-vitro study. Indian J Dent Res. 2010; 21(3):357-9.
13. Mishra P, Mishra S. Study of Antibacterial Activity of Ocimum sanctum Extract Against Gram Positive and Gram Negative Bacteria. Amer J of Food Tech, 2011; 6: 336-341.
14. Sindhu S, Manorama S, Alfamol PM. Preliminary phytochemical analysis and antimicrobial activity of Piper longum L. (Piperaceae) Mintage. J of Pharma & Med Sci. 2013; 2(1): 21-23.
15. Karsha PV, Lakshmi OB. Antibacterial activity of black pepper (Piper longum Linn.) with special reference to its mode of action on bacteria. Ind J of Nat Prod & Res. 2010;1(2): 213-215.
16. Williamson EM. Major herbs of Ayurveda Churchill living stone, Elsevier Science Ltd, London, 2002.
17. Joy B, Sandhya CP, Remitha KR. Comparison and bioevaluation of Piper longum fruit extracts. J. Chem. Pharm. Res. 2010; 2(4):696-706.
18. Singh C, Rai NP. In-vitro Antibacterial Activity of Piper longum L. Fruit. Int. J. Pharm. Sci. Rev. Res. 2013; 18(2): 89-91.
19. Khan M, Siddique. Antimicrobial activity of piper fruits. Natural products radiance 2007; 6(2): 111-113.
20. Molla AH, Zakaria MG , Molla MTH, Alam MT, Ahsan MS. Chemical investigations and microbial activity of Tinospora cordifolia miers. J. Bio. Sci. 2012; 29:153-160.
21. Singh SS, Pandey SC, Srivastava S, Gupta VS, Patro B, Ghosh AC. Chemistry and medicinal properties of Tinospora cordifolia (Guduchi). Indian J. Pharmacol. 2003; 35: 83-91.
22. Alam P, Ali M, Singh R, Madhurima, Ahmad S, Shakeel F. A validated HPLC method for estimation of cordifolioside A in Tinospora cordifolia, Miers and marketed formulations. J Chromatogr Sci. 2009; 47(10):910-3.
23. Jeyachandran R, Xavier TF, Anand SF. Antibacterial activity of stem extracts of Tinospora cordifolia (Willd) Hook. f & Thomson. Anc Sci Life. 2003; 23(1): 40-43.
24. Gahlawat D, Mazumdar A. In-vitro antibacterial activity of the whole plant of Tinospora cordifolia. Int J of Health and Pharma Sci.2013; 2(3): 1-6.
25. Upadhay RK, Tripathi R, Ahmad S. Antimicrobial activity of two Indian medicinal plants Tinospora cordifolia (family Menispermaceae) and Cassia fistula (Family: Caesalpinaceae) against human pathogenic bacteria. J of Phar Res. 2011; 4(1):167-170.
26. Lekouch N, Sedki A, Nejmeddine A, Gamon S. Lead and traditional Moroccan pharmacopoeia. Sci Total Environ. 2001; 280:39-43.
27. Dwivedi SK, Dey S. Medicinal herbs: A potential source of toxic metal exposure for man and animals in India. Arch Environ Health. 2002; 57:229-31.
28. Gajalakshmi S, Iswarya V, Ashwini R, Divya G, Mythili S, Sathiavelu A. Evaluation of heavy metals in medicinal plants growing in Vellore District. Eur J of Exp Biol. 2012; 2(5):1457-1461.
29. Dzomba P, Chayamiti T, Togarepi E. Heavy Metal Content of Selected Raw Medicinal Plant Materials: Implication for Patient Health. Bull. Environ. Pharmacol. Life Sci. 2012; 1(10): 28-33.
30. Sivananthan M .Antibacterial activity of 50 medicinal plants used in folk medicine International Journal of Biosciences 2013; 3(4):104-121.
31. IS 5887 (Part 1). Isolation, Identification and Enumeration of E. coli.1976 (Reaffirmed 2005).
32. IS 14843. Meat and meat products – Enumeration of Pseudomonas spp. 2000.
33. IS 5887 (Part 3). General Guidance on Methods for the Detection of Salmonella. 1999 (Reaffirmed 2005).
34. IS 5887 (Part 7).General Guidance on Methods for the Isolation and Identification of Shigella. 1999 (Reaffirmed 2005).
35. IS 5887(Part 2). Isolation, Identification and Enumeration of S. aureus and F. streptococci. 1976 (Reaffirmed 2005).
36. Debnath M, Chejara V, Vijaya BK, Bika D, Sajal K, Dixit A, Jain R, Jain V. Evaluation of quality and impact of untreated wastewater for irrigation. Amer J Res Com. 2014: 2(4): 200-231.
37. Temminghoff EJM, Houba VJG. Plant analysis procedures. 2nd ed. Dordrecht: Kluwer Academic Publishers. 2004.
38. Joshi B, Lekhak S, Sharma A. Antibacterial Property of Different Medicinal Plants: Ocimum sanctum, Cinnamomum zeylanicum, Xanthoxylum armatum and Origanum majorana. Kat Uni J of Sci Eng and Tech. 2009; 5(1): 143- 150.
39. Bhatt MK, Shankar MB, Saluja AK, Dholwani KK, Captain AD. Evaluation of antimicrobial activity of Ocimum sanctum methanolic extract. J Pharm Sci Innov. 2012; 1(4): 39-41.
40. Abeysinghe PD, Wanigatunge RP. Evaluation of antibacterial activity of different mangrove plant extracts. Ruh J of Sci. 2006; 1:104-112.
41. Kamegam N, Karuppusamy S, Prakash M, Jayakumar M, Rajasekar K. Antibacterial potency and synergistic effects of certain plant extracts against food borne diarrheagenic bacteria. Int J of biomed and pharma sci. 2008:2(2): 88-93.
42. Rivera SEV, Escobar-Saucedo MA, Morales D, Aguilar CN, Herrera RR. Synergistic effects of ethanolic plant extract mixtures against food-borne pathogen bacteria. Afr J of Biotech.2014; 13(5):699-704.
43. Sahu M, Sujatha S, Chaya DR, Parija SC. Pericardial effusion–an unusual manifestation of salmonellosis: a case report. Cases J. 2008; 1:375.
44. Fontaine A, Arondel J, Sansonetti PJ. Construction and evaluation of live attenuated vaccine strains of Shigella flexneri and Shigella dysenteriae. Res Microbiol.1990; 141(7-8):907-12.
45. Lunavath V, Porika R, Priya S , Mamidala E Preliminary phytochemical analysis and anti bacterial activity of Tinospora cordifolia leaf extracts. Inter J of Health and Pharma Sci. 2012; 1(4):18-21.
46. Zhang W, Robertson DC, Zhang C, Bai W, Zhao M, Francis DH. Escherichia coli constructs expressing human or porcine enterotoxins induce identical diarrheal diseases in a piglet infection model. Appl. Environ. Microbiol.2008; 74:5832-5837.
47. Ahn SH, Tsalik EL, Cyr DD, Zhang Y, van Velkinburgh JC, Langley RJ, Glickman SW, Cairns CB, Zaas AK, Rivers EP, Otero RM, Veldman T, Kingsmore SF, Lucas J, Woods CW, Ginsburg GS, Fowler VG Jr. Gene expression-based classifiers identify Staphylococcus aureus infection in mice and humans. PLoS One. 2013; 8(1):e48979.
48. Shamina A, Sarma YR. Secondary metabolites in black pepper (Piper nigrum) and their effect on the foot-rot pathogen Phytophthora capsici. J of Plantation Crops, 2001; 29 (2): 22-26.
49. Singh V, Amdekar S, Verma O. Ocimum sanctum (tulsi): Bio-pharmacological Activities. Pharmacol. 2010; 1(10):WMC001046.
50. Mishra A, Kumar S, Pandey AK. Scientific Validation of the Medicinal Efficacy of Tinospora cordifolia. Scientific World J. 2013(2013); Article 292934.8 pages.
51. WHO: Geneva Switzerland; 1998. Quality control methods for medicinal plant materials. available at http://whqlibdoc.who.int/publications/1998/9241545100.pdf .
52. Nayak P. Aluminum: impacts and disease. Environ Res. 2002; 89(2):101-15.
53. Abdurrahman FI, Tijjani MA, Osuji UO. Proximate content and chemical composition of Ocimum viridis leaf and Ocimum gratissum leaf. Internat Res J of Phar 2012; 3(4): 153-156.
54. Ling LJ, Clark RF, Erickson TB, Trestrail JH. philadelphia: Hanley and Belfus Inc; 2001. Toxicology secrets, 2001, pp. 153-69.
55. Khan MA, Ahmad I, Rahman I. Effect of environmental pollution on heavy metals content of Withania somnifera. J of the Chinese Chem Soc. 2007; 54:339-43.
57. Annan K, Dickson, Amponsah IK, Nooni IK. The heavy metal contents of selected plants sampled from different locations. Pharmacog Res. 2013; 5(2): 103-108
58. Choy CMY, Lam CW, Cheung LT, Briton-Jones CM, Cheung LP, Haines CJ. Infertility, blood mercury concentrations and dietary seafood consumption: A case-control study. BJOG. 2002;109:1121-5.
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01-07-2014
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“EVALUATION OF HEAVY METAL DISTRIBUTION AND ANTIBACTERIAL ACTIVITIES OF MEDICINAL PLANTS TINOSPORA CORDIFOLIA, OCIMUM SANCTUM AND PIPER NIGRUM”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 6, no. 3, July 2014, pp. 229-34, https://doi.org/10.25004/IJPSDR.2014.060312.
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“EVALUATION OF HEAVY METAL DISTRIBUTION AND ANTIBACTERIAL ACTIVITIES OF MEDICINAL PLANTS TINOSPORA CORDIFOLIA, OCIMUM SANCTUM AND PIPER NIGRUM”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 6, no. 3, July 2014, pp. 229-34, https://doi.org/10.25004/IJPSDR.2014.060312.
