Simultaneous Determination of Seven Components from Hawthorn Leaves Flavonoids in Rat Plasma by LC-MS/MS

In this study, a simple, sensitive, and throughout liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of seven flavonoid compounds, namely, rutin, vitexin-4 ''-O-glucoside, vitexin-2 ''-O-rhamnoside, hyperoside, vitexin, shanyenoside A and quercetin in rat plasma after intravenous administration of hawthorn leaves flavonoids (HLF) using lysionotin as an internal standard (IS). The target compounds were extracted using protein precipitation by methanol. The detection was achieved by LC-MS/MS in multiple reaction monitoring mode. The optimal mass transition ion pairs (m/z) for quantitation were 609.3/300.1 for rutin, 593.1/413.2 for vitexin-4 ''-O-glucoside, 577.3/413.2 for vitexin-2 ''-O-rhamnoside, 463.2/300.1 for hyperoside, 431.2/311.2 for vitexin, 407.2/245.1 for shanyenoside A, 301.1/151.1 for quercetin and 343.2/313.1 for the IS, respectively. The method was fully validated with respect to specificity, sensitivity, linearity, precision, accuracy, recovery and stability experiments. A sufficiently sensitive and selective LC-MS/MS method was first developed in this study to simultaneously evaluate the pharmacokinetics of seven flavonoids in rat plasma following intravenous administration of HLF.