Simultaneous quantification of eight constituents in the seed of Forsythia suspense (Thunb.) Vahl by HPLC-MS/MS method / 中草药

ABSTRACT

Objective: To develop an HPLC-MS/MS method for the simultaneous determination of forsythoside A, phillyrin, forsythiaside, rutin, quercetin, isoquercitrin, ferulic acid, and hesperidin in the seed of Forsythia suspense (Thunb.) Vahl. Methods: Chromatographic separation was performed on a Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) with linear gradient elution of methanol and 0.1% formic acid at a flow rate of 800 μL/min, and the injection volume was 10 μL. Multiple-reaction monitoring (MRM) was employed with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at-4 500 V and the turbo spray temperature was maintained at 650℃. Results: All calibration curves showed good linearity within the test ranges. The average recoveries of the compounds ranged from 98.1% to 101.8%. The precisions (RSD) for the investigated components were less than 0.84%. The contents of forsythoside A, phillyrin, forsythiaside and rutin are higher than quercetin, isoquercitrin, ferulic acid and hesperidin. And the average contents of forsythoside A, phillyrin, forsythiaside and rutin are 1 609.78, 80.08, 86.64 and 373.86 μg/g, respectively. Conclusion: An efficient, rapid, and sensitive method is first established for the qualitation and quantification of eight major components in the seed of Forsythia suspense (Thunb.) Vahl. The satisfactory results demonstrate that the method could be applied as a reliable quality control method for the seed of Forsythia suspense (Thunb.) Vahl.