Rapid identification and analysis of the active components of traditional Chinese medicine Xiaoxuming decoction for ischemic stroke treatment by integrating UPLC-Q-TOF/MS and RRLC-QTRAP MSn method.

Xiaoxuming decoction (XXMD) is a key Chinese medicine prescription, which has been clinically used for stroke treatment for thousands of years in ancient China. The extracted active fraction of XXMD (AF-XXMD) contains almost pharmacological active components with anti-cerebral ischemic effects. However, the illumination of its complex ingredients remains challenging. In this study, ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and rapid resolution liquid chromatography-triple quad linear ion trap mass spectrometry (RRLC-QTRAP MSⁿ) methods were developed for the qualitative and quantitative analysis of AF-XXMD, respectively. Data showed that 48 compounds were identified in AF-XXMD by using UPLC-Q-TOF/MS, including 14 alkaloids, 14 flavonoids, 12 triterpenoids, 3 chromones, 3 monoterpenes, 1 cyanide glycoside, and 1 volatile oil. Among them, 38 components were unambiguously characterized by their reference standards. A total of 15 compounds in AF-XXMD were first reported. Additionally, 33 compounds were quantified by using RRLC-QTRAP MSⁿ in AF-XXMD. This developed RRLC-QTRAP MSⁿ method provides an adequate linearity (r² > 0.99) and intrabatch and interbatch variations (RSD < 15%), with recovery (60.3%-107.5%) of 33 compounds concerned. The total content of 33 compounds in AF-XXMD reached 31.53%. The high total contents of compounds of Xing Ren, Shao Yao, and Huang Qin in AF-XXMD were 9.52%, 8.85%, and 7.62%, respectively. The data further showed that cyanophoric glycosides, monoterpenes, and flavonoids were the three most abundant components in AF-XXMD. Results provide advantageous information for the comprehensive study of the pharmacokinetic features and pharmacological mechanisms of AF-XXMD.