DOI: http://dx.doi.org/10.18203/2319-2003.ijbcp20175692

Quantification of rutin and quercetin by HPTLC/HPLC and in vitro immunomodulatory and anticancer activities of Capparis moonii fruits extracts

Gaurav Mahesh Doshi, Manjushree Kundalik Pawar, Kajal Haribhai Chavda

Abstract


Background: The current research was undertaken on dried fruits of Capparis moonii to screen its potential for immunomodulatory and cancer indications with identification of phytoconstituents by chromatographic techniques.

Methods: Methanolic (MECN), hydro-methanolic (HMECN) and aqueous extracts (AQCN) of Capparis moonii were subjected to high performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) after studying the total phenolic and flavonoid content by using rutin and gallic acid as standards respectively as well as undertaking powder characteristics and preliminary phytochemical screening. Immunomodulatory activities covered were hemagglutination antibody titre and delayed-type hypersensitivity reaction with the aid of sheep red blood cells (0.5×109) as antigens.  The extracts were studied for antioxidant potential. Anticancer prospects were focusing on in vitro cell lines screening (MCF 7 and HCT 15) by Sulforhodamine B assay method and potato disc assay.

Results: The total phenolic and flavonoid content of MECM, HMECM and AQCM fruits extracts were found to be 0.20, 0.11 and 0.47 mg of gallic acid/g and 78.3, 18.8 and 64.4 mg of rutin/g respectively. Rutin and quercetin were confirmed by HPTLC and HPLC showing well resolved peaks. IC50 values in antioxidant studies were found to be significant with all the extracts. Significant immunomodulatory effect was noticed at 200mg/kg in both models (high antibody titre levels and decrease paw volume after 48 h). Unsatisfactory results were observed with selected cell lines and disc assay.

Conclusions: Thus, selected fruits may probably have immunomodulatory potential due to presence of flavonols (rutin and quercetin).


Keywords


Capparis moonii, Delayed-type hypersensitivity, Haemagglutination antibody titre, Potato disc assay, Sulforhodamine B assay method

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References


Doshi GM, Une HD. In vitro cytotoxicity studies on Carissa congesta, Polyalthia longifolia, and Benincasa hispida extracts by Sulforhodamine B assay method. Int J Gr Pharm. 2015;9(3):157-61.

Ananthanaryan R, Paniker JCK. Immunodeficiency diseases. In Text of Microbiology Part II. 6th Ed. Orient Longman Limited Publishing House. India; 2000:142-149.

Doshi GM, Une HD. Screening of Polyalthia longifolia Leaves as Potential Immunomodulatory Target. Int J Pharmacol. 2015;11(2):106-13.

Vanita GK, Brijendra J, Pramila Y. Evaluation of in-vitro immunomodulatory activity of aqueous and ethanolic extract of Capparis moonii. Int J Pharm Bio Sci. 2013;4(2):344-52.

Mishra N, Tomar PC, Lakra N. Medicinal and food value of Capparis- a harsh terrain plant. Ind J Trad Knowl. 2007;6(1):230-8.

Khandelwal KR. Practical Pharmacognosy-Techniques and Experiments.10th Ed. Nirali Prakashan Pune. India; 2000:149-56.

Kokate CK. Practical Pharmacognosy. 2nd Ed. India: Vallabh Prakashan, New Delhi; 1989:111-4.

Alali F, Tawaha K, El-Elimat T, Syouf M, El-Fayad M, Abulaila K, et al. Antioxidant activity and total phenolic content of aqueous and methanolic extracts of Jordanian plants: an ICBG project. Nat Prod Res. 2007;21:1121-31.

Doshi GM, Chaskar PK, Zine SP, Une HD. Solicitation of HPLC and HPTLC for determination of rutin from Polyalthia longifolia. Pharmacog Res. 2014;6(3):234-9.

Harborne JB. Phytochemical methods: A guide to Modern Techniques of Plant Analysis. 3rd Ed. Springer (India) Private limited Publication House,New Delhi; 2005:11-12.

Gupta SM, Shivaprasad HN, Kharya MD, Rana AC. Immunomodulatory activity of the Ayurvedic Formulation “Ashwagandha churna”. Pharm Bio. 2006;44:263-5.

Bader JM. Identification of T lymphocytes. In: Glase J C (Eds.) Tested Studies for Laboratory Teaching, Kendall; Hunt Publishing Company, USA;1997(18):113-123.

OECD Guideline for Testing of Chemicals: Acute Oral Toxicity – Acute Toxic Class Method: Adopted: 17th Ed. 2001:1-14.

Shukla S, Mehta A, John J, Mehta P, Vyas SP. Immunomodulatory activities of ethanolic extract of Caesalpinia bonducella seeds. J Ethno pharmacol. 2009;125:252-6.

Vogel GH. In: Drug Discovery and Evaluation, Pharmacological Assays. Chapter I: Methods for testing immunological factors, 2nd Ed. India; 2002:788-820.

Stankovic MS. Total phenolic content, flavonoid concentration and antioxidant activity of Marrubium peregrinum L. extracts. Kragujevac J Sci. 2007;33:63-72.

Skehn P, Storeng R, Scudiero A, Monks J, McMohan D, Vistica D, et al. New colorimetric cytotoxicity assay for anticancer drug screening. J Nat Cancer Ins. 1990;82:1107.

Vanicha V, Kanyawim K. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nature Protocols. 2006;1:1112-6.

Coker PS, Radecke, J, Guy, G, Camper ND. Potato disc tumor induction assay: A multiple mode of drug action assay. Phytomedicine. 2003;10:133-8.

Nagore DH, Patil PS, Kuber, VK. Comparison between High Performance Liquid Chromatography and High Performance Thin Layer Chromatography for determination of Diosgenin from the fenugreek seeds. Int J Gr Pharm. 2012;6(4):315-20.

Barar FSK. In: Essentials of Pharmacotherapeutics. Chapter: Immune System, Immunosuppressants and Immunostimulants. 5th Ed. S Chand and Company Limited, New Delhi; 2009:489-492.

Benacerraf B. A hypothesis to relate the specificity of T lymphocytes and the activity of I region specific IR genes in macrophages and borrower lymphocytes. J Immunol. 1978;120:1809-32.

Goodman and Gilman. In: The Pharmacological Basis of Therapeutics. Edited by Laurence L. Bruton and John S Lozo, Keith L Parker. Chapters: Pharmacotherapy of Asthma by Bradley JU and Immunodulators Immunosuppressants, Tolerogens and Immunostimulants. 11th Ed. Mc-Graw Hill. New Delhi; 2005:717-736.

Miller LE In: Ludke HR, Peacock JE, Tomar RH (Eds.), Lea and Febiger. Manual of Laboratory Immunology. London; 1991:1-18.

Nair PP, Turjman N, Kessie G, Calkins B, Goodman GT, Davidovitz H, et al. Diet, nutrition intake, and metabolism in populations at high and low risk for colon cancer. Am J Clin Nut. 1984;40:927-30.

Doshi GM, Une HD. Carissa congesta Wight and Benincasa hispida (Thunb.) Cogn. as Budding Immunomodulatory Targets. Ind J Exp Bio. 2016;54(10):650-8.

Nautiyal J, Kanwar SS, Yu Y, Majumdar APN. Combination of dasatinib and curcumin eliminates chemo-resistant colon cancer cells. J Mol Sig. 2011;6:1-11.

Parasramka MA, Gupta SV. Synergistic effect of garcinol and curcumin on antiproliferative and apoptic activity in pancreatic cancer cells. J Oncol. 2012;2012:709-39.

Keepers PY, Piazo PE, Peters JG, Otte JVA, Winogard B, Pinnedo HM. Comparison of the Sulforhodamine B protein and Tetrazolium (MTT) Assays for in vitro chemosenstivity testing. Eur J Cancer. 1991;27(7): 897-900.