QUALITY EVALUATION OF A PHYTOPHARMACEUTICAL BY HPTLC AND SCREENING OF ITS IMMUNOSTIMULANT & ANTIMICROBIAL ACTIVITIES

Main Article Content

Madhu. C. Divakar
WD Sam Solomon
Dinesh B
Dharma S

Keywords

Phytopharmaceutical formulation, HPTLC, Ocimum sanctum, Allium cepa, Mentha piperita, Immunostimulant, Antimicrobial activity, Herbovigilance

Abstract

The present work includes the HPTLC quality evaluation of a phytopharmaceutical (immunostimulant & expectorant) formulation prepared from three herbs namely Ocimum sanctum (tulsi), Allium cepa (red onion) and Mentha piperita (mentha) as per the guidelines of Indian Pharmacopoeia Commission, Ministry of Health and family welfare, Govt. of India.  The latest direction of Indian Pharmacopoeia commission indicates for the inclusion of a minimum of four therapeutic or analytical markers in a phytopharmaceutical formulation (PPF). In the present work eugenol, menthol and quercetin were used as the markers for the herbal drugs Ocimum sanctum, Allium cepa and Mentha piperita respectively.


 The Herbovigillance part of the work can give information specific to herb-drug interactions for the patients taking these kinds of preparations along with conventional drugs in the post covid-19 pandemic period.  Herbovigillance protocols were framed for the patients under anti-diabetic drugs, anti-coagulants, anti-allergic drugs, and anti-hypertensives for monitoring carefully if taking the prepared phytopharmaceutical formulation along with these conventional drugs.


The antimicrobial activity studies showed zone of inhibition of 11.8 mm and 10.6mm for E. coli (gram-ve) and Staph.aures (gram +ve) respectively by PPF. The percentage immunostimulation was found to be 64% and 82% for the PPF at the dose of 50mg/ml and100 mg/ml respectively.


The HPTLC analysis of the phytopharmaceutical formulation indicated that, an amount of 0.008 mcg of eugenol (Rf value: 0.83), 0.008 mcg of quercetin (Rf value: 0.73), 0.004 mcg of menthol (Rf value: 0.59) are present per mcg of Ocimum sanctum, Allium cepa and Mentha piperita extracts respectively. The results can give an insight for herbal manufacturers for the quality development of phytopharmaceutical formulations

Abstract 161 | pdf Downloads 22

References

1. The Govemment of India in the Ministry of Health and Family Welfare (Department of Health and Family Welfare), number G.S_R_ 702(E), dated the October, 2013, in the Gazette of India, Extraordinaly, Part 11_ Section 3, Sub-section dated the 24m October, 2013.
2. Bhatt A, Phytopharmaceuticals: A new drug class regulated in India, Perspectives in Clinical Research 2016; 7(2): 59-61.
3. Zulfa Nooreen, Vineet Kumar Rai and Narayan Parasad Yadav, Phytopharmaceuticals: A new class of drug in India, Annals of Phytomedicine 2018;7(1):27-37.
4. Bhusnure OG, Shinde MC, Vijayendra SS, Gholve SB, Giram PS, Birajdar MJ. Phytopharmaceuticals: An emerging platform for innovation and development of new drugs from botanicals, Journal of Drug delivery and therapeutics 2011; 9(3):1046-57.
5. John Dike Nwabueze Ogbonna, Franklin Chimaobi Kenechukwu, Anthony Amaechi Attama, Salome Amarachi Chime, Different approaches to formulation of herbal extracts /phytopharmaceuticals /bioactive phytoconstituents - a review, International Journal of Pharmaceutical Sciences Review and Research, Int. J. Pharm. Sci. Rev. Res.2012;16(1):1-8.


6. The Gazette of India, Extraordinary Part II-Section 3- Sub-section(i), Published by Authority, New Delhi, Monday, November 30,2015/ AGRAHAYANA 9,1937, MINISTRY OF HEALTH AND FAMILY WELFARE (Department of Health and Family Welfare), Notification, New Delhi, the 30th November,2015 G.S.R. 918(E).
7. Cimpoiu C, Jantschi L, Hodisan, Journal of T.T Planar chromatogram 1998; 703(1-2) 11:191-4.
8. Meena. S. Deogade, Prasad KSR, Standardization of wild Krishnatulsi (Ocimum tenuiflorum Linn), International Journal of Ayurvedic medicine 2019;10(1):52-61.
9. Mondal S, Bijay R, Mirdha BR, Mahapatra SC, The science behind sacredness of Tulsi (Ocimum.), Indian Journal of Physiology and Pharmacology 2009;53(4): 291-306.
10. Pawan kumar Ahirwar, Jyotsana Pathak, Prabhat Soni, Manoj Kumar Tripathi, Pharmacognostical standardisation and HPTLC fingerprint of various species of Ocimum leaf, Journal of emerging technologies and innovative research 2019; 6(1): 295-301.
11. R.R. Chattopadhyay. Hypoglycaemic effect of Ocimum sanctum leaf extract in normal and Streptozotocin diabetic rats. Indian J Exp Biol. 1993; 31(11):891-3.
12. Boullata J. Natural health product interactions with medication. Nutr Clin Pract. 2005; 20: 33-51.
13. Datta D, Chandola H, Agarwal SK, Shukla VJ, Pandya PN. Pharmacognostical and analytical study of Tulsi-Amla-Yasti Ghrita. Ayu 2012; 33(2):274-278.
14. Wang An, SavasUzen, Stout C. David and Johnson Eric F. Structural Characterization of the Complex between -Naphthoflavone and Human Cytochrome P450-1B1. Journal of Biological Chemistry 2010; 5736-5743.
15. Basheer A. Al-Sum, Abdullah A. Al-Arfaj. Antimicrobial Activity of the Aqueous Extract of Mint. Plant Science Journal of Clinical Medicine 2013; 2(3): 110-113.
16. G. Bupesh, C. Amutha, S. Nandagopal, A. Ganeshkumar, P. Suresh kumar, K. Saravana Murali. Antibacterial activity of Mentha piperita L. (peppermint) from leaf extracts – a medicinal plant. Acta agriculturae Slovenica 2007; 89 (1): 73–79.
17. Okmen AS, Okmen G, Arslan A, Vurkun M. Antibacterial Activities of Mentha piperita L. Extracts Against Bacteria Isolated from Soccer Player’s Shoes and its Antioxidant Activities. Indian J of Pharmaceutical Education and Research 2017;5 1(3): S163-69.
18. Hubert Antolak, Agata Czy˙zowska and Dorota Kr˛egiel. Activity of Mentha piperita L. Ethanol Extract against Acetic Acid Bacteria Asaia spp. Foods 2018; 7(171): 1-10.
19. Jehan Bakht, Shehla Khan and Mohammad Shafi. Antimicrobial Potentials of Fresh Allium Cepa Against Gram Positive and Gram-Negative Bacteria and Fungi. Pak. J. Bot. 2013; 45(S1): 1-6.
20. Veronika Bugarova, Jana Godocikova, Marcela Bucekova, Robert Brodschneider and Juraj Majtan. Effects of the Carbohydrate Sources Nectar, Sucrose and Invert Sugar on Antibacterial Activity of Honey and Bee-Processed Syrups. Antibiotics 2021; 10: 985.
21. Intan Putri Hapsaria and Yoanni Maria Lauda Feroniasanti. Phytochemical Screening and in Vitro Antibacterial Activity of Sweet Basil Leaves (Ocimum basilicum L.) Essential Oil against Cutibacterium acnes ATCC11827,1-7.
22. Rakesh Kumar Joshi, William N Setzer and Joyce Kelly da Silva. Phytoconstituents, traditional medicinal uses and bioactivities of Tulsi (Ocimum sanctum Linn.): A review American Journal of Essential Oils and Natural Products 2017; 5(1): 18-21.
23. Kaushik Vilas Kulkarni and Dr. Belvotagi Venkatrao Adavi Rao. A review on: Indian traditional shrub Tulsi (Ocimum sanctum): The unique medicinal plant. Journal of Medicinal Plants Studies 2018; 6(2): 106-110.
24. Bridson E.Y., Natural and Synthetic Culture Media for Bacteria. Hand book series in Nutrition and Food section 6, Vol. III Ed. Recheigl, CRC Press, Ohio, 1978; 91-281.
25. Kulkarni SR, Karnade VS, Study of immunopotentiating activity of naphthoquinione extracts of leaves of Lawsonia alba. Indian drugs 1983; 35(7): 427-433.
26. Farnsworth NR. Biological and Phytochemical screening of plants. Int.J. Pharm.Sciences 1966; 55:225-76.
27. Indian Pharmacopoeia 2022, vol 3, Govt.of India, Ministry of health and family welfare, published by Indian Pharmacopoeia commission, Ghaziabad, 4310, 4252,4263.
28. WHO monographs on selected medicinal plants. Vol. 2. Geneva, World Health Organization, 1999, 16,188, 206.
29. Divakar. C. Madhu., (2021) Plant Drug Evaluation-A laboratory guide, CD remedies, Cochin, 4th Edn, 18,84,195. DOI: 10.15254/PDE.2015

Most read articles by the same author(s)