UPLC快速测定葛根芩连汤肠外翻囊样品中6个黄酮类成分含量

目的: 建立UPLC法同时测定葛根芩连汤肠外翻囊样品中6个黄酮类成分(葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷)的含量。 方法: 样品用甲醇提取,采用Agilent Proshell 120 EC-C₁₈(4.6 mm×50 mm,2.7 μm)色谱柱,以0.2%醋酸水溶液(A)-甲醇(B)为流动相进行梯度洗脱,流速1 mL·min^-1,紫外检测波长270 nm,柱温30 ℃。 结果: 葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷质量浓度分别在0.1870～93.50 μg·mL^-1(r=0.9999),0.02730～13.65 μg·mL^-1(r=0.9999),0.02550～12.75 μg·mL^-1(r=0.9999),0.05580～27.90 μg·mL^-1(r=0.9998),0.2124～106.2 μg·mL^-1(r=0.9999),0.09140～45.70 μg·mL^-1(r=0.9999)范围内有良好的线性关系;日间、日内精密度变异均小于2%;平均回收率(n=9)分别为101.4%,97.88%,99.15%,99.72%,98.50%,101.4%;稳定性良好。 结论: 所建立的分析方法操作简便、快速、灵敏,结果准确,重复性好,所需样品少,并已成功应用于葛根芩连汤大鼠肠吸收动力学的研究。

Objective: To develop a sensitive and selective ultra performance liquid chromatography(UPLC) method for identification and quantification of six flavonoids components(puerarin, daidzin, liquiritin, scutellarin, baicalin, wogonoside) in rat everted gut sacs. Methods: The method was based on simple liquid-liquid extraction. The separation was performed on a Agilent Proshell 120 EC-C₁₈(4.6 mm×50 mm, 2.7 μm) column.The mobile consisted of 0.2% acetic acid aqueous(A) and methanol(B) with a gradient elution at a flow-rate of 1.0 mL·min^-1. The detection wavelength was 270 nm and the column temperature was 30 ℃. Results: The linear ranges of puerarin, daidzin, liquiritin, scutellarin, baicalin, wogonoside were 0.1870-93.50 μg·mL^-1(r=0.9999),0.02730-13.65 μg·mL^-1(r=0.9999),0.02550-12.75 μg·mL^-1(r=0.9999),0.05580-27.90 μg·mL^-1(r=0.9998),0.2124-106.2 μg·mL^-1(r=0.9999),0.09140-45.70 μg·mL^-1(r=0.9999), respectively. The intra-and inter-day precisions(RSD) were within 2% for the analytes. The mean recoveries of puerarin, daidzin, liquiritin, scutellarin, baicalin, wogonoside were 101.4%, 97.88%, 99.15%, 99.72%, 98.50%, 101.4%. Conclusion: The proposed method enables unambiguous identification and quantification for the preliminary pharmacokinetic studies of the compounds(puerarin, daidzin, liquiritin, scutellarin, baicalin, wogonoside), and has been applied to a intestinal absorption kinetics of Gegenqinlian decoction in rats successfully.