In Vitro Callus Culture of Centella Asiatica L. (Urb.) and its HPLC Analysis

C. M. Shashikala¹*, S. Shashidhara² and P. E. Rajshekharan²

¹Department of Pharmacognosy, Govt. College of Pharmacy, Bangalore India

²Indian Institute of Horticulture Research, Bangalore India.

Corresponding Author E-mail: shashimetri@rediffmail.com

ABSTRACT: Different explants of Centella asiatica were excised and cultured on Murashige & Skoog (MS) media supplemented with different growth hormones at different concentrations. It has been found that the frequency of callus induction was more on MS media supplemented with Naphthalene acetic acid (NAA) (0.5 mg-1) + kinetin (2.0 mg-1) and NAA (0.5mg-1) + Benzyl Amino Purine (BAP) (2.0 mg-l). Among the leaf, petiole and root explants, petiole explants showed more frequency of callus induction followed by leaf and root explants. Qualitative chemical tests of the methanolic extracts of callus showed the presence of saponins, triterpenes, flavonoids but the absence of alkaloids and qualitative analysis by thin layer chromatography (TLC), ultra violet (UV) and infra red (IR) analysis confirm the presence of asiaticoside. Quantitative High performance liquid chromatography (HPLC) analysis reveals that the leaf callus contains significantly more amounts of asiaticoside (1.1%) and madecassoside (2.4%)) compared to the root and petiole callus on dry weight basis.

KEYWORDS: Centella asiatica; callus culture; asiaticoside; HPLC

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