Quantitative and rapid quality assessment methods for the multi‐class bioactive constituents of Tinospora cordifolia using high‐performance liquid and thin layer chromatography analysis with tandem mass spectrometry characterization

Abstract

Tinospora cordifolia is an important traditional botanical medicine used as an immunomodulator with diversified therapeutic potential. The present study used the systematic isolated Tinospora cordifolia compounds (tinosporide, 8‐hydroxy tinosporide, tinosporaside, cordifolioside A, 20‐β‐hydroxy ecdysone, and columbin) with four alkaloids (syringin, magnoflorine, tetrahydropalmatine, and jatrorrhizine) to develop and validate a high‐performance liquid chromatography method for simultaneous quantification of these multi‐class bioactive. This novel method showed good specificity with excellent linearity (r2 > 0.99), precision (relative standard deviation < 5.0%), and robust and accurate performance (93.68–99.76%) for 10 Tinospora cordifolia compounds as an effective strategy for quantification using the single and multi‐class standards approach. The rapid marker‐based qualitative high‐performance thin‐layer chromatography methods were also developed to analyze samples as a visual quality control tool. These methods were first applied to analyze 15 Tinospora cordifolia samples using the external standard method. By analyzing the results based on the multi‐class (%relative error 2.19–10.56) and single standard (%relative error 25.00–70.50) approach, the multi‐class standard assessment can be effectively adapted for Tinospora cordifolia samples. The quantitative‐qualitative analysis was further confirmed with mass spectroscopy characterization. This multi‐faceted analytical approach provides a unique solution for fingerprint, quality assurance, and identification of Tinospora cordifolia with tinosporide, 8‐hydroxy tinosporide, and tinosporaside as an authenticated marker.