高效液相色谱法测定紫苏中5种有机酸和黄酮的含量

Determination of Five Organic Acids and Flavonoids in Perilla frutescens by High Performance Liquid Chromatography

目的:建立以甲醇溶液为萃取剂,结合超声辅助萃取,利用高效液相色谱法同时分离测定紫苏中的咖啡酸、阿魏酸、迷迭香酸、木犀草素和芹菜素5种有效成分的方法。方法:采用Phenomenex C18色谱柱(250 mm×4.6 mm,5μm);流动相为甲醇-0.1%冰醋酸溶液,采用梯度洗脱程序;流速1 mL/min;紫外检测波长320 nm;柱温35℃。结果:该方法对咖啡酸、阿魏酸、迷迭香酸、木犀草素和芹菜素分别在0.001 22~61.00(r=0.999 99)、0.001 06~53.00(r=0.999 99)、0.001 28~64.00(r=0.999 91)、0.001 20~60.00(r=0.999 94)、0.001 12~56.00μg/mL(r=0.999 95)范围内线性关系良好,相关系数(r)均大于0.999 91,检出限(RSN=3)依次是:0.101 9、0.392 6、0.355 9、0.286 2、0.202 5 ng/mL;样品回收率为95.79%~101.41%。结论:本法操作简单快速、定量准确、灵敏度高、成本低、且对环境友好,为紫苏中咖啡酸、阿魏酸、迷迭香酸、木犀草素和芹菜素的分离检测提供了一个有效的方法。

Objective: A simple high performance liquid chromatography(HPLC) method has been developed for the simultaneous quantification of caffeic acid,ferulic acid,rosmarinic acid,luteolin and apigenin from Perilla frutescens using methanol as extractant with ultrasonic-assisted extraction. Methods: The five compounds were isolated by using a Phenomenex C18 column by gradient elution,and the mobile phase was methanol-0.1% acetic acid solution with a flow rate of 1.0 mL/min. The UV detection wavelength was performed at 320 nm. Results: The linear ranges for caffeic acid,ferulic acid,rosmarinic acid,luteolin and apigenin were determined to be 0.001 22–61.00,0.001 06–53.00,0.001 28–64.00,0.001 20–60.00 and 0.001 12–56.00 μg/mL,respectively,and the correlation coefficients were 0.999 99,0.999 99,0.999 91,0.999 94 and 0.999 95,respectively. The limits of detection(LOD) were 0.101 9,0.392 6,0.355 9,0.286 2 and 0.202 5 ng/mL,respectively. The average recoveries of these compounds in spiked real samples were in the range of 95.79%–101.41%. Conclusion: The proposed method is simple,precise,specific,sensitive and accurate. It provides a new and scientific means for routine quality control of Perilla frutescens.