• Deep Chhavi Anand Tissue culture and Biotechnology laboratory, Department of Botany, University of Rajasthan, Jaipur-302004
  • Rishikesh Meena Tissue culture and Biotechnology laboratory, Department of Botany, University of Rajasthan, Jaipur-302004
  • Vidya Patni Tissue culture and Biotechnology laboratory, Department of Botany, University of Rajasthan, Jaipur-302004


Objective: The aim of the present study was to develop a callus induction protocol and comparative study of therapeutic phytochemicals present in in vivo leaf and in vitro callus extracts through Gas Chromatography-Mass Spectrometry analysis.

Methods: Murashige and Skoog media was used as culture media for callus induction. In vitro callus induction protocol was developed by studying the effects of various plant growth regulators like auxin, 2, 4-D (2,4-dichlorophenoxyacetic acid), NAA (naphthalic acetic acid), alone and in combination with cytokinin BAP (benzyl aminopurine), on leaf and stem explants. The GC-MS analysis of Ampelocissus latifolia was carried out on Shimadzu QP-2010 plus with thermal desorption system TD 20 to study the phytochemical profile.

Results: In vitro callus induction protocol was developed for the plant and callusing was done from leaf and stem explants of Ampelocissus latifolia. The best result for callus induction was obtained using leaf explant, and callus production were maximum in Murashige and Skoog medium fortified with BAP (0.5 mg/l) and NAA (1.0 mg/l). Major compounds identified in the GC-MS analysis were Campesterol, Stigmasterol, Beta-Sitosterol, Docosanol, Dodecanoic acid, etc., in in vitro extract and Beta Sitosterol, Tocopherol, Squalene, Bergamot oil, Margarinic acid, Hexadecanoic acid, etc., in in vivo extract. The different active phytochemicals identified have been found to possess a wide range of biological activities, thus this analysis forms a basis for the biological characterization and importance of the compounds identified for human benefits.

Conclusion: This is the first report on callus induction in Ampelocissus latifolia. From the results obtained through the in vitro callus induction and its comparative GCMS analysis with in vivo extract, it is revealed that Ampelocissus latifolia contains various bioactive compounds that are of importance for phytopharmaceutical uses. The GCMS analysis revealed that the amount of Beta-sitosterol and 5-Hydroxymethylfurfural (HMF) was very high in in vitro extract as compared to in vivo extract.

Keywords: Ampelocissus latifolia, GC-MS, Growth regulators, Stigmasterol, Callus, Phytochemicals


Download data is not yet available.


1. Maheswari J. Patenting Indian medicinal plants and products. Indian J Sci Technol 2011;4:298-301.
2. Nandagoapalan V, Doss A, Marimuthu C. Phytochemical analysis of some traditional medicinal plants. Bioscience Discovery 2016;7:17-20.
3. Jiang W, Myeong Je C, Lemaux PG. Improved callus quality and prolonged regenerability in the model and recalcitrant barley (Hordeum vulgare L.) cultivars. Plant Biotechnol 1998;15:63–9.
4. Anand DC, Patni V. Ampelocissus latifolia (Roxb.) Planch.–a traditional plant with enormous medicinal and economic importance. Int J Pharma Biosci 2016;7:303-7.
5. Tamilarasi CT, Subasini U, Kavimani S, Jaykar B. Phytochemical and pharmacological evaluation of Ampelocissus latifolia. Ancient Sci Life 2000;20:1-6.
6. Choudhury S, Choudhury HR, Mandal S. Pharmacognostic studies of Ampelocissus latifolia (Roxb.) Planch.–An Important Ethnomedicinal Plant. Bhubaneswar. Proceedings of the 99th session of the Indian Science Congress (II): Section XIV: Plant Sciences; 2012. p. 123.
7. Patil MV, Patil DA. Some herbal remedies used by the tribals of Nasik district, Maharashtra. Natl Prod Radiance 2007;6:152-7.
8. Chauhan A, Goyal MK, Chauhan P. GC-MS technique and its analytical applications in science and technology. J Anal Bioanal Tech 2014;5:222.
9. Ronald Hites A. Gas chromatography-mass spectroscopy: a handbook of instrumental techniques for analytical chemistry; 1997. p. 609-11.
10. Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962; 15:473–97.
11. Pednekar PA, Kulkarni V, Raman B. Physicochemical and phytochemical evaluation of Ampelocissus latifolia (Roxb.) planch leaves. Int J Pharm Pharm Sci 2014;6:504-7.
12. Sundararaman P, Djerassi C. A convenient synthesis of progesterone from stigmasterol. J Org Chem 1977;42:3633-4.
13. Kametani T, Furuyama H. Synthesis of vitamin D3 and related compounds. Rev Article 1987;7:147-71.
14. Aringer L, Eneroth P, Nordstrom L. Side chain hydroxylation of cholesterol, campesterol and β-sitosterol in rat liver mitochondria. J Lipid Res 1976;17:263–72.
15. Awad AB, Chen YC, Fink CS, Hennessey T. β-sitosterol inhibits HT-29 human colon cancer cell growth and alters membrane lipids. Anticancer Res 1996;16:2797–804.
16. Genser B, Silbernagel G, De Backer G, Bruckert E, Carmena R, Chapman MJ, et al. Plant sterols and cardiovascular disease: a systematic review and meta-analysis. Eur Heart J 2012;33:444-51.
17. Sacks SL, Thisted RA, Jones TM, Barbarash RA, Mikolich DJ, Ruoff GE, et al. Clinical efficacy of topical docosanol 10% cream for herpes simplex labialis: a multicenter, randomized, placebo-controlled trial. J Am Acad Dermatol 2001;45:222-30.
18. Marounek M, Putthana V, Benada O, Lukesova D. Antimicrobial activities of medium-chain fatty acids and monoacylglycerols on Cronobacter sakazakii DBM 3157T and Cronobacter malonaticus DBM 3148. Czech J Food Sci 2012;30:573–80.
19. Yang D, Pornpattananangkul D, Nakatsuji T, Chan M, Carson D, Huang CM, et al. The antimicrobial activity of liposomal lauric acids against Propionibacterium acnes. Biomaterials 2009;30:6035-40.
20. Aranceta J, Perez Rodrigo C. Recommended dietary reference intakes, nutritional goals and dietary guidelines for fat and fatty acids: a systematic review. Br J Nutr 2012;107:S8–S22.
21. Berger NA. Obesity and cancer pathogenesis. Ann N Y Acad Sci 2014;1311:57–76.
22. Gopala Krishnan S. GC-MS analysis of some bioactive constituents of Mussaenda frondosa Linn. Int J Pharma Biosci 2011;2:313-20.
23. Huang Zih Rou, Lin Yin Ku, Fang Jia You. Biological and pharmacological activities of squalene and related compounds: potential uses in cosmetic dermatology. Molecules 2009;14: 540–54.
24. Romano L, Battaglia F, Masucci L, Sanguinetti M, Posteraro B, Plotti G, et al. In vitro activity of bergamot natural essence and furocoumarin-free and distilled extracts, and their associations with boric acid, against clinical yeast isolates. J Antimicrob Chemother 2005;55:110–4.
25. Rizvi S, Raza ST, Ahmed F, Ahmad A, Abbas S, Mahdi F. The role of vitamin e in human health and some diseases. Sultan Qaboos Univ Med J 2014;14:157-65.
26. Potawale SE, Luniya KP, Mantri RA, Mehta UK, Sadiq MW, Vetal YD. Chenopodium ambrosioides: an ethnopharmacological review. Pharmacol Online 2008;2:272-86.
27. Abdulmalik O, Safo MK, Chen Q, Yang J, Brugnara C, Ohene-Frempong K, et al. 5-hydroxymethyl-2-furfural modifies intracellular sickle haemoglobin and inhibits sickling of red blood cells. Br J Haematol 2005;128:552–61.
28. Zhao L, Chen J, Su J, Li L, Hu S, Li B, et al. In vitro antioxidant and antiproliferative activities of 5-hydroxymethylfurfural. J Agric Food Chem 2013;61:10604-11.
29. Li YX, Li Y, Qian ZJ, Kim MM, Kim SK. In vitro antioxidant activity of 5-HMF isolated from marine red alga Laurencia undulata in free-radical-mediated oxidative systems. J Microbiol Biotechnol 2009;19:1319–27.
30. Venkata Raman B, Samuel LA, Pardha Saradhi M, Narashimha Rao B, Naga Vamsi Krishna A, Sudhakar M, et al. Antibacterial, antoxidant activity and GC-MS analysis of Eupatorium odoratum. Asian J Pharm Clin Res 2012;5:99-106.
197 Views | 591 Downloads
How to Cite
Anand, D. C., R. Meena, and V. Patni. “IN VITRO CALLUS INDUCTION AND COMPARATIVE GC-MS ANALYSIS OF METHANOLIC EXTRACTS OF CALLUS AND LEAF SAMPLES OF AMPELOCISSUS LATIFOLIA (ROXB.) PLANCH”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 9, Sept. 2018, pp. 68-72, doi:10.22159/ijpps.2018v10i9.27879.
Original Article(s)