ESTIMATION OF SOME SECONDARY METABOLITES FROM THE IN VITRO CULTURES OF CHLOROPHYTUM BORIVILIANUM SANT. Et. FERN
Keywords:Chlorophytum borivilianum, Phytochemical, ESIMS, GCMS, In vitro cultures
Objective: To perform phytochemical screening, estimate total saponins and to study the secondary metabolite profiling of Chlorophytum borivilianum by ESI-MS (Electron spray ionisation-mass spectroscopy) and GC-MS (Gas chromatography-mass spectroscopy) techniques.
Methods: The powdered samples of leaves and roots (nature grown and freshly harvested and hardened in vitro regenerated) and callus of Chlorophytum borivilianum (2 g) were extracted exhaustively by soxhlet by refluxing with petroleum ether (60-80 Â°C) and then with ethanol (85%). Alternatively, the methanol extract was prepared for the samples by the cold maceration method and the filtered extract was used. This extract was further analyzed by gas chromatography-mass spectrometry to identify and characterize the chemical compounds present in the crude extracts. Vanillin sulphuric acid assay was done to estimate the total saponin content of extracts.
Results: Phytochemical analysis of ethanolic extract showed the presence of major classes of phytochemicals. The spectrophotometric analysis as well as the GC-MS results revealed the remarkably high percentage of saponin content in the in vitro roots. Besides this, many important secondary metabolites viz. Î²-Sitosterol and Taraxerone were also revealed by GC-MS.
Conclusion: Results of this study showed enhanced production of many important secondary metabolites, especially saponins in tubers of in vitro regenerated plantlets vis a vis that of the nature grown plants establishing the fact that micropropagation can pave new vistas for the research in phytopharmaceuticals and bioprospecting of rare and important medicinal plants.Â
Namdeo AG. Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev 2007;1:69-79.
Singh A, Chauhan HS. Safed musli (Chlorophytum borivilianum): distribution, biodiversity and cultivation. J Med Aromat Plant Sci 2003;25:712-9.
Kothari SK. Safed musli (Chlorophytum borivilianum) revisited. J Med Aromat Plant Sci 2004;26:60-3.
Thakur M, Dixit VK. Fructan; the polymer with unexplored potential. Indian Pharm 2005;4:7-12.
Sreevidya N, Govindarajan R, Vijayakuma M, Thakur M, Dixit VK, Mehrotra S, et al. Action of fructo-oligo polysaccharide fraction of Chlorophytum borivilianum against streptozotocin induced oxidative stress. Planta Med 2006;72:1421-4.
Thakur M, Dixit VK. A review on some important medicinal plants of Chlorophytum spp. Pharmacogn Rev 2008;2:168â€72.
Purohit SS, Prajapati ND. Local heritage with global importance. AGROBIOS News Lett 2003;1:7-8.
Kemat N, Kadir MA, Abdullah NAP, Ashraf F. Rapid multiplication of safed musli (Chlorophytum borivilianum) through shoot proliferation. Afr J Biotechnol 2010;9:4595-4600.
Maiti S, Geetha KA. Safed Musli. Published by indian council of agricultural research (ICAR) New Delhi; 2009. p. 18â€39.
Dave A, Bilochi G, Purohit SD. Scaling-up production and field performance of micropropagated medicinal herb â€˜safed musliâ€™ (Chlorophytum borivilianum). In vitro Cell Dev Biol: Plant 2003;39:419-24.
Basu S, Jha T. In vitro propagation of the rare medicinal plant Chlorophytum borivilianum Sant et. Fern. In: Proc Nat Symp of Plant Biotechnology; 2007. p. 285-9.
Rizvi MZ, Kukreja AK, Knanuja SPS. In vitro culture of Chlorophytum borivilianum Sant et Fernand in liquid culture medium as a cost-effective measure. Curr Sci 2007;92:87-90.
Jha A, Bansal YK. High frequency direct somatic embryogenesis from the juvenile shoot buds of Chlorophytum borivilianum Sant. Et Fernand and production of artificial seeds. Int J Pharm Sci Res 2016;7:2040-9.
Obadoni BO, Ochuko PO. Phytochemical studies and comparative efficacy of crude extracts of some Homostatic plants in Edo and Delta states of Nigeria. Global J Pure Appl Sci 2010;8:203-8.
Haque R, Saha S, Bera T. A peer reviewed of general literature on Chlorophytum borivilianum commercial medicinal plant. Int J Drug Dev Res 2011;3:140â€“55.
Harborne JB. Twenty-five years of chemical ecology. Nat Prod Rep 2001;18:361-79.
Evans WC. In: Evans WC, Trease GE. editors. Trease and Evansâ€™ Pharmacognosy. 14th ed. London: WB Saunders company ltd; 1996. p. 612.
Hiai S, Oura H, Nakajima T. Color reaction of some sapogenins with vanillin and sulfuric acid. Planta Med 1976;29:116â€“22.
Hamburger M, Slacanin I, Hostettmann K, Dyatmiko W. Acetylated saponins with molluscicidal activity from Sapindus rarak: Unambiguous structure determination by proton nuclear magnetic resonance and quantitative analysis. Phytochem Anal 1992;3:231-7.
Li R, Zhou Y, Wu Z, Ding L. ESIâ€“QTOF-MS/MS and APCI-IT-MS/MS analysis of steroid saponins from the rhizomes of Dioscorea panthaica. J Mass Spectrom 2006;41:1-22.
Pascual ME, Carretero ME, Slowing KV, Villar A. Simplified screening by TLC of plant drugs. Pharm Biol 2002;40:139-43.
Fidrianny I, Winarsih S, Ruslan K. Phytochemical content and antioxidant potential of different organs of eggplant (Solanum melongena l.) grown in west Java-Indonesia. Asian J Pharm Clin Res 2017;10:144-9.
Paramasivam I, Simran Sinsinwar S, Muthuraman MS. Phytopharmacological properties of Melothria maderaspatana: a review. Asian J Pharm Clin Res 2017;10:34-6.
Ramachandra Rao S, Ravishankar GA. Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 2002;20:101-53.
Muir AD, Ballantyne KD, Hall TW. LCMS and LC-MS/MS analysis of saponins and sapogeninsâ€“comparison of ionization techniques and their usefulness in compound identification. In: Saponins in Food, Feedstuffs and Medicinal Plants, Annual Proceedings of the Phytochemical Society; 2000. p. 35â€“42.
Schopke T. Non-NMR methods for structure elucidation of saponins. In: Saponins in Food, Feedstuffs and Medicinal Plants. Annual Proceedings of the Phytochemical Society; 2000. p. 106â€“13.
Yang DJ, Lu TJ, Hwang LS. Isolation and identification of steroidal saponins in Taiwanese yam cultivar (Dioscorea pseudojaponica Yamamota). J Agric Food Chem 2003; 51:6438-44.
Kapur S, Pal A, Gurjar S, Pavan VVS, Sharad S. Isolation and characterization of secondary metabolites saponinsâ€ from the explants of Indian medicinal plant Chlorophytum borivilianum. Int J Pharm Front Res 2011;1:14-22.
Gao YH, Mei QX, Wu HF, Diao YM, Peng XS, Zhang RF. Chlorophytoside A, a new labdane diterpene glycoside from Chlorophytum laxum. Chin Chem Lett 2005;16:925-7.
Acharya D, Mitaine-Offer AC, Kaushik N, Miyamoto T, Lacaille Dubois MA. Furostane-type steroidal saponins from the roots of Chlorophytum borivilianum. Helvetica Chim Acta 2008;91:2262-9.
Sharma A, Suchitra M, Anirudha R. Quantitative estimation of Î²-sitosterol and stigmasterol in vivo and in vitro Terminalia chebula Ritz. Int Res J Pharmacol 2012;2:115-6.
Kulkarni DD, Ghugale DD, Narsimhan R. Chemical investigation of plant tissues grown in vitro: isolation of beta-sitosterol from Morus alba (mulberry) callus tissue. Indian J Exp Biol 1970;8:347.
Migas P, Luczkiewicz M, Cisowski W. The influence of auxins on the biosynthesis of isoprene derivatives in callus cultures of Vaccinium corymbosum var. blue crop. Zeitschrift Naturforschung 2006;61:565-70.
Sharma N, Nathawat RS, Gour K, Patni V. Establishment of callus tissue and effect of growth regulators on enhanced sterol production in Cissus quadrangularis L. Int J Pharmacol 2011;7:653-8.
Bathoju G, Giri A. Production of stigmasterol and hecogenin from in vitro cultures of Chlorophytum borivilianum. J Pharmacog 2012;3:101-3.
Piironen V, Lindsay DG, Miettinen TA, Toivo J, Lampi AM. Plant sterols: biosynthesis, biological function and their importance to human nutrition. J Sci Food Agric 2000;80:939-66.
Dufourc EJ. The role of phytosterols in plant adaptation to temperature. Plant Signal Behav 2008;3:133-4.
Shaik S, Singh N, Nicholas A. HPLC and GC analyses of in vitro grown leaves of cancer bush Lessertia frutescens (Sutherlandia) reveals higher yields of bioactive compounds. Plant Cell Tiss Organ Cult 2011;105:431-8.
Roja PC, Heble MR. Indole alkaloids in clonal propagation of Rauvolfia serpentina benth ex. Kurz. Plant Cell Tiss Organ Cult 1996;44:111-5.
Guo B, Gao M, Liu CZ. In vitro propagation of an endangered medicinal plant Saussurea involucrate Kar. Et Kir. Plant Cell Rep 2007;26:261-5.