EXTRACTION, Extraction, Isolation, Identification and Estimation of Diosgenin by TLC Profiling and UHPLC-LC-SRM Analysis in Three Costus Species

Medicinal plants are utilized for treating many ailments, since ancient times due to their tremendous therapeutic properties. The plant derivatives are used as a raw material for the synthesis of drugs either as a natural or artificial synthetic drug. Costaceae family members are known to possess many medicinal properties few of them are mainly consumed to reduce the blood glucose level and treat many disorders. The sapogenin contained in the rhizome of three Costus species was extracted, identified, and characterized. Young rhizomes were harvested surface sterilized, hydrolyzed; extracted through a soxhlet apparatus and concentrated fractions were subjected to a preliminary phytochemical screening test for saponin, Thin Layer Chromatography, and the UHPLC-MS SRM analysis. The result showed that the maximum yield of the Diosgenin was obtained from the chloroform extract of Costus speciosus. The pharmaceutical analysis in all the three Costus species carried out with isolated saponin was compared with standard Diosgenin. The characterization of isolated diosgenin was done by UHPLC MS and SRM, based on the retention time of all three samples and the comparison with standard Diosgenin. Isolated Diosgenin was quantified through thin layer chromatography with n-hexane: ethyl acetate (7:3) as mobile phase at Rf value of 0.48. The amount of Isolated Diosgenin was estimated by comparing the peak area at a retention time Rt value of 8.5 in all the samples and compared with standard Diosgenin high amount of Diosgenin was found in Costus speciosus rhizome followed by Costus igneus and Costus pictus. The preliminary phytochemical screening and analytical methods involved during the present study were found satisfactory. Thus, the protocol can be used for the extraction of Diosgenin and further used in the synthesis of Drugs. Keywords: Costus pictus, Costus speciosus, Costus igneus, rhizome, TLC, UHPLC-MS, SRM