Identification and characterization of berberine in tinospora cordifolia by liquid chromatography quadrupole time of flight mass spectrometry (LC MS/MS Q-Tof) and evaluation of its anti inflammatory potential

Abstract

Introduction: Tinospora cordifolia (Willd.) Miers ex Hook F and Thomas commonly called as gudduchi or amrita is a widely used plant in traditional medicinal system of Ayurveda. A UPLC MS/MS Q-Tof method for the identification and characterization of berberine in Tinospora cordifolia (Willd.) Miers. ex HooK.F. and Thomas. and to evaluate the anti inflammatory potential of bioactive fraction. Materials and Methods: The presence of berberine in Tinospora cordifolia was determined by HPLC and was subsequently isolated by HPTLC. The anti inflammatory property of the fraction containing berberine was demonstrated to have an inhibitory activity on 5 lipoxygenase, an enzyme involved in inflammatory pathway and its IC50 value was obtained. The binding interactions between berberine and 5-LOX were demonstrated by docking studies. Result: The presence of berberine in Tinospora cordifolia methanolic extract was identified by HPLC and HPTLC analysis and confirmed by UPLC MS/ MS Q-Tof. The fraction containing berberine inhibited 5-LOX with an IC50 of 0.041± 0.0003μg/mL as compared to that of NDGA (positive control) which showed an IC50 of 2.75 ± 0.05 μg/mL. Molecular docking of berberine with 5-LOX showed a binding energy of -8.942 ± 0.039665 kcal/mol and Ki of 273.16 ± 3.026 nM as compared to the NDGA which has a binding energy of -7.186 ± 0.170503 kcal/mol and Ki 5.604± 1.618 μM. Conclusion: Tinospora cordifolia can be used as a source of berberine and possible anti inflammatory activity of Tinospora cordifolia may be attributed to the presence of berberine.