Development and validation of analytical methods for the simultaneous estimation of biologically active selected herbal constituents in some plant extracts and their commercial formulations by HPLC and LC MS techniques

Abstract

A simple, specific, accurate and precise high performance liquid chromatographic and liquid chromatography with mass spectroscopic methods were developed for the simultaneous estimation of flavonoid constituents from Aganosma dichotoma and Achillea millefolium plants. METHOD: The chromatographic conditions such as the type of the stationary phase, nature and composition of the mobile phase, peak modifiers, flow rate, pressure used, detection wavelength, selection of mass range (LC-MS), linearity range, retention and selectivity, efficiency of separation and sensitivity were optimized. Standard solutions were prepared and injected using the optimized chromatographic conditions. The chromatograms were recorded. A calibration curve was plotted by taking the selected analyte concentration on X axis and the peak area of the standard chromatogram on Y axis. RESULT: The proposed method was validated in terms of the analytical parameters such as accuracy, linearity, precision, robustness, limit of detection, limit of quantification were determined based on the International Conference on Harmonization (ICH) guidelines. The HPLC and LC-MS methods developed for the simultaneous estimation of the flavonoids showed that the results were consistent. CONCLUSION: The newly developed methods can be used in various fields like research institutions, quality control department in industries, approved testing laboratories etc. Hence the present RP-HPLC and LC-MS methods were suitable for the quality control of the raw materials, extracts and assay of the markers in Achillea millefolium and Aganosma dichotoma plants and in the formulations.