Characterization of Tinospora Cordifolia accessions from different districts of Himachal Pradesh

Abstract

newline ABSTRACT newlineTinospora cordifolia commonly named as AMRITA which is attributed to this wonder drug for its ability to impart youthfulness, longeveity and vitality to its patron, is one of the potent immunomodulator. Present study encompasses (i) genetic diversity of T.cordifolia (ii) antimicrobial potency (iii) secondary metabolite makeup. The study was undertaken for evaluating the genetic variation within 10 accessions of T. cordifolia collected from different Districts of Himachal Pradesh. Analysis was made using 18 decamer primers. Out of them, 9 primers showed reproducible distinct polymorphism patterns. Maximum genetic diversity was observed in the sample collected from Baijnath village of District Kangra. Considering the vast potentiality of T. cordifolia as a source for antimicrobial drugs with reference to antibacterial, antifungal and anticandida agents, a systematic investigation was undertaken to screen 17 samples of T. cordifolia collected from different Districts of Himachal Pradesh for its activity against gram positive and gram negative bacteria, fungus and Candida strains. Acetonic, methanolic and chloroformic extracts were tested but acetonic leaf extract of T. cordifolia collected from Baijnath village of District Kangra showed the maximum inhibitory activity against tested pathogens. Further, phytochemical screening was performed for qualitative determination of the phytoconstituents in the acetonic leaf extract of T. cordifolia. FT-IR analysis of the acetonic leaf extract revealed the presence of different functional groups at wave numbers corresponding to the functional groups of Berberine. Thin layer chromatography and HPLC of acetonic leaf extract of T. cordifolia indicated alkaloid as a major active compound. The Rf value of 0.23 in TLC chromatogram and HPLC peak at Rt 4.024 minutes further confirms the presence of Berberine in the acetonic leaf extract. Active compound i.e Berberine was isolated and purified by crysyallization method and tested for its antibacterial activity. The isolated compound was characterized by HPLC, LCMS, GCMS, H1 and C13 NMR. HPLC peak of isolated Berberine at retention time 4.024 mins was observed which was same as that of standard Berberine. Molar mass observed by LCMS and GCMS was 336.1 which was same as that of standard Berberine. NMR confirms the presence of 18 H atoms as that present in standard Berberine. The present study reveals the antibacterial potency correlated with genetic diversity and characterization of Berberine compound from T. cordifolia which would be of immense value in botanical identification and authentication of plant drug.