A UFLC-MS/MS method for simultaneous quantitation of spinosin, mangiferin and ferulic acid in rat plasma: application to a comparative pharmacokinetic study in normal and insomnic rats.

Suan-Zao-Ren (SZR) decoction, consisting of Ziziphi Spinosae Semen, Poria, Chuanxiong Rhizoma, Anemarrhenae Rhizoma and Glycyrrhizae Radix Et Rhizoma, is a Traditional Chinese Medicine prescription, clinically used for the treatment of insomnia. The objective of this study was to develop a sensitive and reliable UFLC-MS/MS method for simultaneous quantitation of spinosin, mangiferin and ferulic acid, the main active ingredients in SZR decoction, and to compare the pharmacokinetics of these active ingredients in normal and insomnic rats orally administrated with the prescription. Analytes and IS were separated on a Shim-pack XR-ODS column (75 mm × 3.0 mm, 2.2 µm particles) using gradient elution with the mobile phase consisting of methanol and 0.1% formic acid in water at a flow rate of 0.4 mL/min. The detection of the analytes was performed on 4000Q UFLC-MS/MS system with turbo ion spray source in the negative ion and multiple reaction-monitoring mode. The lower limits of quantification were 1, 6 and 1 ng/mL for spinosin, mangiferin and ferulic acid, respectively. Intra- and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). The mean extraction recoveries of analytes and IS from rats plasma were all more than 85.0%. The validated method has been successfully applied to comparing pharmacokinetic profiles of analytes in rat plasma. The results indicated that no significant difference in pharmacokinetic parameters of ferulic acid was observed between two groups, while absorptions of spinosin and mangiferin in insomnic group were significantly lower than those in normal group.