PHYTOCHEMICAL ANALYSIS AND EVALUATION OF IN VITRO IMMUNOMODULATORY ACTIVITY OF RHODODENDRON ARBOREUM LEAVES
DOI:
https://doi.org/10.22159/ajpcr.2018.v11i8.25372Keywords:
Rhododendron arboreum, Phytochemical, Ursolic acid, Kaempferol, Immunomodulatory activityAbstract
Objectives: The objectives of this study are to analyze phytoconstituents in Rhododendron arboreum leaves (qualitatively and quantitatively) and to estimate the immunomodulatory effects of extracts using different in vitro methods for intracellular and phagocytic killing potency of neutrophils, which are subsequent, involved in the process of phagocytosis by neutrophils.
Methods: R. arboreum leaves extracts were separately extracted in 100% methanol, 50% methanol, and aqueous by a hot percolation technique using Soxhlet apparatus and concentrated with a vacuum rotary evaporator apparatus. The percentage yield of the extracts wares calculated. The extract was used to analyze phytoconstituents qualitatively, and total phenolic and flavonoid content were estimated. In another study, already purified methanolic extract of R. arboreum leaves, namely cytoplasmic male sterility (CMS-3) (kaempferol), RAM fr2, and transcranial magnetic stimulation (TMS-10) (ursolic acid) was used to evaluate immunomodulatory activities by nitroblue tetrazolium test, phagocytosis of killed Candida albicans.
Results: Phytochemical screening of the extracts showed the presence of major classes of phytochemicals, i.e., phytosterols, flavonoids, phenols, alkaloids, carbohydrate, glycoside, sterols and steroids, terpenoids, and tannin while saponin was absent in all extract. The percent yield of leaves extract was 12.97%. Total phenolic content (TPC) expressed as gallic acid equivalents (GAE) ranged 1327.5±0.009 (in 100% methanolic extract), 1309.5±0.011 (in 50% methanolic), and 1468.5±0.006 mg GAE g−1 dry weight (in aqueous extract). Total flavonoid content expressed as rutin equivalents (RE) ranged 219±0.025 (in 100% methanolic extract), 184±0.02 (in 50% methanolic), and 262±0.58 mg (in aqueous extract) mg RE g−1 dry weight. R. arboreum purified extract CMS-3 (kaempferol), RAM fr2, and TMS-10 (ursolic acid) stimulated the neutrophils to phagocytic activity to the extent of 67.67±2.08%, 40.3±1.5%, and 79.67±0.57%, respectively. The above-said extracts stimulated the phagocytosis of killed C. albicans. The mean particle numbers for CMS- 3, RAM fr2, and TMS-10 were found to be 6.7, 6.3, and 7, respectively, at concentration 1 mg/ml.
Conclusion: The phytochemical screening of all the extracts showed the presence of various phytochemicals that are biologically important. The total phenolic and flavonoid content of the plant are comparable to other medicinal plants. Even though the enormous progress on the phytochemistry and pharmacology of R. arboreum have been made, there still require more conclusive studies on the safety, efficacy, and in vivo toxicity of extracts and pure compounds to gain a better understanding.
Downloads
References
Anaissie EJ, Stratton SL, Dignani MC, Lee CK, Summerbell RC, Rex JH, et al. Pathogenic molds (including Aspergillus species) in hospital water distribution systems: A 3-year prospective study and clinical implications for patients with hematologic malignancies. Blood 2003;101:2542-6.
Njoroge GN, Bussmann RW. Diversity and utilization of antimalarial ethnophytotherapeutic remedies among the Kikuyus (Central Kenya). J Ethnobiol Ethnomed 2006;2:8.
Wagner H. Structural analysis of a rhamnoarabinogalactan and arabinogalatans with immunostimulating activity from Calendula officinalis. Photochemistry 1999;28:2379-83.
van der Nat JM, Klerx JP, van Dijk H, de Silva KT, Labadie RP. Immunomodulatory activity of an aqueous extract of Azadirachta indica stem bark. J Ethnopharmacol 1987;19:125-31.
Sohni YR, Bhatt RM. Activity of a crude extract formulation in experimental hepatic amoebiasis and in immunomodulation studies. J Ethnopharmacol 1996;54:119-24.
Kulkarni SR, Karande VS. Study of the immunostimulant activity of napthoquinone extract of leaves of Lawsonia alba linn. Indian Drugs 1998;35:427-33.
Biswas K, Chattopadhyay I, Banerjee RK, Bandyopadhyay U. Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci 2002;82:1136-345.
Chiang LC, Ng LT, Chiang W, Chang MY, Lin CC. Immunomodulatory activities of flavonoids, monoterpenoids, triterpenoids, iridoid glycosides and phenolic compounds of Plantago species. Planta Med 2003;69:600-4.
Kuo YC, Yang LM, Lin LC. Isolation and immunomodulatory effect of flavonoids from Syzygium samarangense. Planta Med 2004;70:1237-9.
Ravichandiran V, Vishal P. In vitro evaluation for immunomodulatory activity of Acorus calamus on human neutrophils. Int Res J Pharm 2015;6:450-2.
Ponkshe CA, Indap MM. In vivo and in vitro evaluation for immunomodulatory activity of three marine animal extracts with reference to phagocytosis. Indian J Exp Biol 2002;40:1399-402.
Daniel PS. In: Daniel PS, Abba IT, Tristram GP, editors. Basic and Clinical Immunology. 8th ed. USA: Appleton and Lange; 1994. p. 195 215.
Orwa C, Mutua A, Kindt R, Jamnadass R, Simons A. Agroforestree Database: A Tree Reference and Selection Guide Version; 2009;4.0.
Rai T, Rai L Tress of the Sikkim Himalaya. New Delhi: Indus Publishing Company; 1994. p. 94.
Chauhan NS. Medicinal and Aromatic Plants of Himachal Pradesh. New Delhi: Indus Publishing Company; 1999. p. 353.
Nisar M, Ali S, Qaisar M. Antibacterial and cytotoxic activities of the methanolic extracts of Rhododendron arboreum. J Med Plants Res 2013;7:398-403.
Prakesh V, Rana S, Sagar A. Studies on antibacterial activity of leaf extracts of Rhododendron arboreum and Rhododendron campanulatum. Int J Curr Microbiol Appl Sci 2016;5:315-22.
Saranya D, Ravi R. The leaf of Nilgiri Rhododendron: A potent antimicrobial agent against medically critical human pathogens. Int J Pharmacog 2016;3:251-56.
Chauhan P, Singh J, Sharma RK, Easwari TS. Anti-bacterial activity of Rhododendron arboreum plant against Staphylococcus aureus. Indian J 2016;9:92-6.
Painuli S, Rai N, Kumar K. Gas chromatography and mass spectrometry analysis of methanolic extract of leaves of Rhododendron arboreum. Asian J Pharm Clin Res 2016;9:101-4.
Prakash D, Upadhyay G, Singh BN, Dhakarey R, Kumar S, Singh KK. Free radical scavenging activities of Himalayan Rhododendrons. Curr Sci 2007;92:526-32.
Prakash T, Fadadu SD, Sharma UR, Surendra V, Goli D, Stamina P, et al. Hepatoprotective activity of leaves of Rhododendron arboreum in CCl4 induced hepatotoxicity in rats. J Med Plants Res 2008;2:315-20.
Sharma UR, Surendra V, Jha SK, Nitesh SC, Prakash T, Goli D. Evaluation of anti-inflammatory activity of Rhododendron arboreum herb extract on experimental animal. Arch Pharm Res 2009;1:58-61.
Verma N, Singh AP, Amresh G, Sahu PK, Rao CV. Antiinflammatory and antinociceptive activity of Rhododendron arboreum. J Pharm Res 2010;3:1376-80.
Verma N, Singh AP, Amresh G, Sahu PK, Rao CV. Protective effect of ethyl acetate fraction of Rhododendron arboreum flowers against carbon tetrachloride-induced hepatotoxicity in experimental models. Indian J Pharmacol 2011;43:291-5.
Srivastava P. Rhododendron arboreum: An overview. J Appl Pharm Sci 2012;2:158-62.
Sonar KP, Singh R, Bansal P, Balapure KA, Saraf KS. R. arboreum flower and leaf extracts: RP-HPTLC screening, isolation, characterization and biological activity. Rasayan J Chem 2012;5:165-72.
Sonar PK, Singh R, Verma A, Saraf SK. Rhododendron arboreum (Ericaceae): Immunomodulatory and related toxicity studies. Orient Pharm Exp Med 2013;13:127-31.
Bhandari L, Rajbhandari M. Isolation of quercetin from flower petals, estimation of total phenolic, total flavonoid and antioxidant activity of the different parts of Rhododendron arboreum smith. Sci World 2014;12:34-40.
Roy JD, Handique AK, Barua CC, Talukdar A, Ahmed FA, Barua IC. Evaluation of phytoconstituents and assessment of adaptogenic activity in vivo in various extracts of Rhododendron arboreum (leaves). Indian J Pharm Biol Res 2014;2:49-56.
Nisar M, Ali S, Muhammad N, Gillani SN, Shah MR, Khan H, et al. Antinociceptive and anti-inflammatory potential of Rhododendron arboreum bark. Toxicol Ind Health 2016;32:1254-9.
Sindiri KM, Machavarapu M, Vangalapati M. Extraction, modeling and purification of flavonoids from Zephyranthes candida. J Int Acad Res Multidiscipl 2013;1:50-6.
Nisar M, Ali S, Qaisar M. Preliminary phytochemical screening of flowers, leaves, bark, stem and roots of Rhododendron arboreum. Middle East J Sci Res 2011;10:472-6.
Kiruba S, Mahesh M, Nisha SR, Miller P. Phytochemical analysis of flower extracts of Rhododendron arboretum sm. ssp. Nilagiricum tagg. Asian Pac J Trop Biomed 2011;S:284-6.
Saklani S, Chandra S. Evaluation of in vitro antimicrobial activity, nutritional profile and phytochemical screening of Rhododendron rboreum. World J Pharm Pharm Sci 2015;4:962-71.
Chandrashekar R, Rao SN. Phytochemical analysis of ethanolic extract of leaves of Lucasindica (Eelli). Int J Pharm Bio Sci 2013;4:33-8.
Chintalapani S, Swathi MS, Narasu ML. Phytochemical screening and in vitro antioxidant activity of whole plant extracts of Sesuvium portulacastrum L. Asian J Pharm Clin Res 2018;11:322-7.
Mathew S, Khosla KK, Mathew C, Bhowmik D. Preliminary phytochemical studies of Kalachoepinnata (Lam) Pers. J Med Plant Stud 2013;1:19-23.
Tariq AL, Reyaz AL. Phytochemical analysis of Camellia sinensis leaves. Int J Drug Dev Res 2012;4:311-6.
Peach K, Tracey MV. Modern Methods of Plant Analysis. Berlin Heidelberg: Springer Verlag; 1956. p. 125-7.
Ahuja J, Suresh J, Deep A, Pratyush M, Ravi. Phytochemical screening of aerial parts of Artemisia parviflora Rox b: A medicinal plant. Scholar Res Lib 2011;3:116-24.
Philip D, Kaleena PK, Valivittan K, Kumar CP. Phytochemical screening and antimicrobial activity of Sansevieria roxburghiana Schult and Schult. F Middle East Jof Sci Res 2011;10:512-8.
Mirela K, Vlasta P, Hribar J, Simcic M. Total phenol content and antioxidant activity of water solutions of plant extracts. Croat J Food Sci Technol 2009;1:1-7.
Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdicphosph-tungstic acid reagents. Am J Enol Vitic 1965;16:144-58.
Kumar P, Pardhu G, Shruti R, Vijya L, Tejaswini K, Vishwanath PB. Phytochemical screening, antimicrobial and anti proliferative properties of Rhododendron ponticum on prostate epithelial cancer cells. J Pharm Sci Res 2011;3:1552-5.
Fidrianny I, Nadia E, Ruslan KW. In vitro antioxidant activities, total flavonoid, phenolic and carotenoid content from various extracts of four species Asteraceae herb. Int J Pharm Pharm Sci 2015;7:192-7.
Dae OK, Seung WJ, Chang YL. Antioxidant capacity of phenolic phytochemicals from various cultivars of plums. Food Chem 2003;81:321-6.
Patil JK, Jalalpure SS, Hamid S, Ahirrao RA. In-vitro immunomodulatory activity of extracts of Bauhinia vareigata Linn stem bark on human neutrophils. Iran J Pharm Ther 2010;9:41-6.
Yadav S, Bhargav M, Ramya R, Laureen, Habeeb N. In-vitro antioxidant and immunomodulatory activity of Citrullus lanatus seed. Int J Eng Sci Res Technol 2016;679-85. DOI: 10.5281/zenodo.212024.
Parameshwara C, Jalalpure SS, Jagwani S, Kumar N, Shah B. Phytochemical investigation and in-vitro immunomodulatorystudy of Asparagus racemosus wild roots extracts. Pharm Sci Monitor 2015;6:292-9.
Kupati VB, Jadar PG. Immunomodulatory effects of ayurvedic drugs: A review. Indian J Appl Res 2014;4:28-30.
Published
How to Cite
Issue
Section
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.
