The integrated study on the chemical profiling and in vivo course to explore the bioactive constituents and potential targets of Chinese classical formula Qingxin Lianzi Yin Decoction by UHPLC-MS and network pharmacology approaches.

Qingxin Lianzi Yin Decoction (QXLZY), a Chinese classical formula, has been widely used in the treatment of various chronic kidney diseases over 1,000 years. However, the current studies on QXLZY were mostly focused on its clinical efficacy, lacking systematic material basis research on constituents. This work aims to elucidate and quantify the chemical constituents, clarify the blood-absorbed components and excretion pathways, predict major bioactive constituents and discover potential therapeutic targets. UHPLC-LTQ-Orbitrap HRMS was employed to clarify the chemical constituents and metabolites of QXLZY. The extraction of diagnostic ion and neutral loss fragment was aimed for searching specific type of constituents. The plasma, urine, bile and feces samples of rats after oral administration of QXLZY were systematically studied. UHPLC-QQQ-MS/MS was employed to simultaneously detect different types of constitutes. Based on the analysis of ingredients in vivo , the bioactive constituents and potential therapeutic targets in the treatment of diabetic nephropathy (DN) was investigated by using network pharmacological analysis. Totally, 220 compounds were identified or tentatively characterized by UHPLC-LTQ-Orbitrap HRMS. Among them, 59 compounds were confirmed by reference standards. Meanwhile, 21 representative components were simultaneously determined within 15 min by UHPLC-QQQ-MS/MS. 123 components (74 prototypes as well as 49 metabolites) were identified or tentatively characterized. By using network pharmacological analysis, baicalein, liquiritigenin, succinic acid, formononetin, wogonin might be the major effective constituents in QXLZY during the treatment of DN. Flavonoids, saponins and organic acids were the major chemical ingredients of QXLZY. Flavonoids were the main components absorbed into blood, followed by organic acids. Phase II conjugation reaction was the major metabolic type. The pathways that QXLZY in the treatment of DN were probably related to glucose and lipid metabolism, oxidative stress and inflammation. By using UHPLC-LTQ-Orbitrap HRMS, 220 compounds were identified or tentatively characterized in Qingxin Lianzi Yin Decoction (QXLZY), a Chinese classical formula. Meanwhile, 21 representative components were simultaneously determined within 15 min by UHPLC QQQ MS/MS. The plasma, urine, bile and feces samples of rats after oral administration of QXLZY were systematically studied. By using network pharmacological analysis, we predicted major bioactive constituents and discovered potential therapeutic targets. [Display omitted] • This study builds a strategy for systematic studying the chemical constituents and predicts mechanism of action of QXLZY. • The uses of MCI gel column and dynamic exclusion method could obtain more information of minor compounds. • Totally, 220 compounds from QXLZY and 123 metabolites from biological samples were characterized by UHPLC-LTQ-Orbitrap MS. • The quantification analysis of 21 constituents in 16 batches of QXLZY samples was tested by UHPLC QQQ MS/MS. • The bioactive constituents and potential therapeutic targets were investigated by network pharmacological analysis. [ABSTRACT FROM AUTHOR]