HPLC Coupled with Chemometric Analysis and LC-MS Studies of Three Flavonoids in Tephrosia purpurea (L.) Pers Revealed Impact of Chemodiversity on the Quest for the Chemical Markers.

Abstract

Globally, Tephrosia purpurea (L.) Pers is used as an important component in herbal drug formulations for liver health. The present study is aimed to develop a suitable analytical approach for simultaneous analysis of three flavonoids (rutin, deguelin and rotenone) to establish quality control methods for plant. A novel High-performance liquid chromatography photodiode array detector (HPLC-PDA) method has been developed to quantify these flavonoids in T. purpurea. The method was validated, and data were subjected to chemometric analysis to select most optimal marker compound. The method that was found linear with R2 values ranges from 0.996 to 0.998 with good recoveries. Intra- and inter-day precision values were <2. HPLC analysis revealed high level of chemodiversity. Quantity of all the three chemical markers was found significantly disparate in samples from different locations. Deguelin was detectable only in three out of total eight samples. However, liquid chromatography-mass spectrometry analysis was found sufficiently sensitive to detect all the compounds in all samples. Thus, results suggest to apply combination of approaches to enhance confidence in chromatographic methods for quality control of herbal drugs. Principal component analysis ranked the markers as Rutin>Rotenone>Deguelin. This comprehensive approach employing multichromatography platforms can be successfully utilized in analysis of these bioactive markers and routine standardization of herbal material and formulations containing T. purpurea.