HPLC FINGERPRINTING OF ETHNOMEDICINAL ANTITUBERCULOSIS PLANTS

Abstract

Modern phytochemical analysis in terms of fingerprinting of different bioactive phytoconstituents in the roots of Leptadenia reticulata and whole herb of Cocculus hirsutu belonging to the family Ascelpidiaceae and Menispermaceae respectively. High-performance liquid chromatography (HPLC) analysis of roots of Leptadenia reticulata and whole herb of Cocculus hirsutus were carried out to develop the fingerprint and verify the presence of standards such as Gallic acid, Catechin, Rutin, Quercetin and Kaempferol in the plant extract. RP-HPLC was performed on an Agilent 1260 Infinity equipped with Inertsil ODS 3V (250 mm x 4.6 mm), 5 µm as a stationary phase, by injecting 20 µL of the sample (1µg/mL) at the flow rate 1.5 mL/min and temperature 30⁰C using mixture of Mobile Phase A consist of 1 % Ortho phosphoric acid in water and mobile phase B consist of acetonitrile (20:80) with 60 min run time and the spectra were measured at 364nm using UV detector. The retention time of standard Gallic acid, Catechin, Rutin, Quercetin and Kaempferol were 6.4 min, 10.8 min, 18.2 min, 24.3 min and 29.1 min respectively. Retention time of roots of Leptadenia reticulata and whole herb of Cocculus hirsutus extracts were similar to that the standard retention time. HPLC fingerprinting shows the overlay between the four different markers such as catechin, Quercetion, Kaemferol and Rutin with aqueous extract of L. reticulata and C. hirsutus. In L. reticulata all the four markers are present and in C. hirsutus contains all three markers except Catechin in given quantity of sample.