目的:建立白花蛇舌草有效部位中对香豆酸和反式6-O-对香豆酰鸡屎藤苷甲酯含量的测定方法。方法:采用高效液相色谱法,色谱柱采用InertsilODS-3(250 mm×4.6 mm,5μm);检测波长为310 nm;进样量为10μL;流速为1.0 mL.min-1;柱温为30℃;以乙腈为流动相A,以0.05%甲酸水溶液为流动相B,进行梯度洗脱。结果:对香豆酸在0.003 33～0.099 9μg范围内成线性关系(r=0.999 9,n=5),平均回收率为99.21%(n=6);反式6-O-对香豆酰鸡屎藤苷甲酯在0.020 9～0.627μg范围内成线性关系(r=0.999 9,n=5),平均回收率为99.54%(n=6)。结论:本实验方法可靠,操作简便,重现性好,专属性强,可用于白花蛇舌草有效部位中对香豆酸和反式6-O-对香豆酰鸡屎藤苷甲酯的含量测定,以控制白花蛇舌草有效部位的质量。 OBJECTIVE: To establish a method for the determination of p-coumaric acid and trans-6-O-p-coumaroyl fetidin methyl ester in the effective part of Hedyotis diffusa. Methods: InertsilODS-3 (250 mm × 4.6 mm, 5 μm) was used as the chromatographic column. The detection wavelength was 310 nm. The injection volume was 10 μL. The flow rate was 1.0 mL.min-1. The column temperature Was 30 ℃; acetonitrile as mobile phase A, 0.05% formic acid aqueous solution as mobile phase B, gradient elution. The results showed that the average recovery of p-coumaric acid was 99.21% (n = 6) in the range of 0.003 33 ~ 0.099 9 μg (r = 0.999 9, n = 5) The results showed that there was a linear relationship (r = 0.999 9, n = 5) in the range of 0.020 9-0.627 μg with an average recovery of 99.54% (n = 6). Conclusion: The experimental method is reliable, easy to operate, reproducible, and specific, and can be used for p-coumaric acid and trans-6-O- Content determination, to control the quality of the effective parts of Hedyotis diffusa.