School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, Liaoning Province, P. R. China.
Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, Liaoning Province, P. R. China.
J Sep Sci. 2017 May;40(9):1896-1905. doi: 10.1002/jssc.201601420. Epub 2017 Apr 19.
An accurate, rapid, and reliable ultra high performance liquid chromatography with tandem mass spectrometry method was developed and validated for the simultaneous determination of baicalin, wogonoside, baicalein, wogonin, and oroxylin A in rat plasma. Then, the stability of baicalin and baicalein in the preparation of plasma sample was systematically investigated. The Waters BEH C column was used with a gradient mobile phase system of acetonitrile and water containing 0.1% formic acid. The analytes were detected in the multiple reaction monitoring mode with positive electrospray ionization. 100 μL fresh plasma was added with 50 μL antioxidant reagent (1 mol/L HCl containing 0.5% Vitamin C), and liquid-liquid extraction with ethyl acetate was used to extract the analytes from plasma. Lower limits of quantification of baicalin, wogonoside, baicalein, wogonin, and oroxylin A were 21.9, 4.80, 1.20, 0.848, and 0.800 ng/mL, respectively. The mean extract recoveries of five flavonoids were 69.1∼89.2%, and the precision and accuracy were within the acceptable limits. This method was further successfully applied to the comparative pharmacokinetic study of these five flavonoids in rats after oral administration of Xiaochaihutang and three compatibilities. The obtained results may be helpful to reveal the mechanism of Xiaochaihutang formula compatibility.
建立并验证了一种用于同时测定大鼠血浆中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素和木蝴蝶苷 A 的准确、快速和可靠的超高效液相色谱-串联质谱法。然后，系统研究了黄芩苷和黄芩素在血浆样品制备过程中的稳定性。采用 Waters BEH C 柱，以含 0.1%甲酸的乙腈和水为梯度流动相系统。采用正电喷雾电离多反应监测模式检测分析物。取 100 μL 新鲜血浆，加入 50 μL 抗氧化剂（含 0.5%维生素 C 的 1 mol/L HCl），用乙酸乙酯进行液-液萃取，从血浆中提取分析物。黄芩苷、黄芩素、汉黄芩苷、汉黄芩素和木蝴蝶苷 A 的定量下限分别为 21.9、4.80、1.20、0.848 和 0.800 ng/mL。五种黄酮类化合物的平均提取回收率为 69.1∼89.2%，精密度和准确度均在可接受范围内。该方法进一步成功应用于大鼠口服小柴胡汤及其三种配伍后五种黄酮类化合物的比较药代动力学研究。所得结果可能有助于揭示小柴胡汤配方配伍的机制。
