In Vitro Propagation, Development and Validation of HPTLC Method for Quantification of Steroidal Glycoalkaloid Solasodine in In Vitro Culture and Plant Parts of Solanum Nigrum

Abstract

Solanum nigrum Linn. (Family: solanaceae), has medicinal values, commonly exploited for its active constituents which are of high pharmaceutical importance. The plant contains different type of natural compounds, and one of most important glycoalkaloid in S. nigrum is solasodine. Nodal segment of S. nigrum was cultured on MS media added with several concentrations of 2, 4-dichlorophenoxy acetic acid (2, 4- D) (0.5ppm –2ppm) for callus induction and development. For shoot induction, MS medium complemented with different dilutions of 6- benzyl adenine (6- BA) (0.5ppm–2ppm) was used. A novel, rapid, and simple high performance thin layer chromatographic method for quantitative estimation of triterpenoid glycoalkaloid. Solasodine was spotted on silica gel 60F254 TLC plate using chloroform: methanol (8.75: 1.25; v/v) as mobile phase. Anisaldehyde-sulfuric acid in methanol used as derivatizing reagent and quantified by densitometric study at 366 nm. This scheme was found to give a sharp and compact spot for the standard drug (Rf = 0.67 ± 0.04). The linearity of this method was found to be a range of 100-1000 ng per spot with LOD and LOQ of 23.78 ng per spot and 67.09 ng per spot; respectively. This method has shown recoveries of solasodine in the range from 98.23% -100.88 %, with low intra and inter-day % RSD. Statistical investigation of the data presented that the method is selective, accurate, precise and reproducible for the study of solasodine. The established and developed HPTLC method would be a significant parameter for the analysis of quality control method of herbal drugs and formulations.