Development and validation of a simple ultra‐high‐performance‐tandem mass spectrometry method for the simultaneous determination of five bioactive components in rat plasma of Hedysari Radix

Abstract

Hedysari Radix is a commonly used traditional Chinese medicine that improves immunity; formononetin, ononin, calycosin, medicarpin, and vanillic acid play an important role in achieving this effect. Herein, a precise and sensitive ultra‐high‐performance‐tandem mass spectrometry method was developed and validated for the determination of five bioactive components that were extracted from plasma using a protein precipitation method. A Waters CORTES C18 (4.6 × 50 mm, 2.7 µm) and a mobile phase composed of methanol and water (containing 0.2% formic acid) were used for the separation. The method was linear within the concentration range of 19.53–625.00 ng/mL for formononetin; 0.01–3.13 ng/mL for ononin; 0.01–3.13 ng/mL for calycosin; 0.16–5.00 ng/mL for medicarpin; and 19.53–625.00 ng/mL for vanillic acid. This method has a lower limit of quantification, is simple, and has a short analysis time and a high degree of separation. The intra‐day precisions and inter‐day precisions of quality control samples were traced to be below 8.58% and 12.64%, respectively. Fidelity intervened from −8.61% to 10%. Finally, the developed method was used to determine the concentrations of the five bioactive components in rat plasma after the administration of rubbing‐ and non‐rubbing processed Hedysari Radix.

Rapid determination of four flavonoids from hawthorn leaves in plasma by ultra‐high performance liquid chromatography‐tandem mass spectrometry and study on its pharmacokinetics

“…Currently, the main research methods of flavonoids in hawthorn leaves include high-performance liquid chromatography (HPLC) [17,18], HPLC-mass spectrometry (HPLC-MS) [19], ultra-high performance liquid chromatography-tandem MS (UHPLC-MS/MS) [20], etc. Among them, UHPLC-MS/MS is widely used in the analysis of pharmacodynamic components and pharmacokinetic studies of traditional Chinese medicine due to its high resolution, high sensitivity, and high accuracy [21,22].…”