Integrating metabolic characterization and pharmacokinetics to uncover the active compounds of <i>Salvia miltiorrhiza-Ligusticum chuanxiong</i> for treating cerebral ischemia

Salvia miltiorrhiza (SM) and Ligusticum chuanxiong (LC) are a well-known herb pair, frequently used in China to treat cerebral ischemia. However, there has been no comprehensive analysis on the metabolic profile, active compounds, and pharmacokinetics of SM−LC. Hence, this work aimed to reveal the active ingredients of SM-LC by establishing its metabolic profile and muti-compound pharmacokinetics by analysis. The strategy integrated visual basic for applications (VBA)-based background removal with a modified mass defect filter (MMDF) technique for the metabolome profiling of SM-LC. A total of 154 exogenous (33 prototypes and 121 metabolites) compounds were quickly characterized in vivo . The predominant metabolic pathways of SM-LC included phase I reactions (methylation, oxidation, hydrogenation, dehydration and hydroxylation, etc.) and phase II reactions (acetylation, glucuronidation and glycosylation, etc.). Then the validated ultra-high performance liquid chromatography connected with tandem triple quadrupole mass spectrometry was employed for the plasma pharmacokinetic analysis of 16 prototypes from SM-LC. Compared with the individual herbs, SM-LC could enhance the C max and area under the curve (AUC) of danshensu, lithospermic acid, senkyunolide I, butylidenephthalide, tanshinone IIA ( P &lt; 0.05). Moreover, neuroprotective activity evaluation indicated that phenolic acids were good at protecting BEND.3 cells from oxygen glucose deprivation/reperfusion (OGD/R) injury, while phthalides and tanshinones exerted obvious neuroprotective effects on HT22 and BV2 cells under the OGD/R injury model. This study would enhance the understanding of the biotransformation profiles and active ingredients of SM-LC against cerebral ischemia, thereby promoting the clinical application of related prescriptions.