HPLC法测定秦皮4种有效成分的含量

HPLC method for simultaneous determination of four component contents in Fraxini Cortex

目的采用高效液相色谱法建立秦皮药材中秦皮甲素、秦皮苷、秦皮乙素、秦皮素的含量测定方法,为建立秦皮饮片质量评价体系提供含量测定方法学依据。方法制备秦皮供试品溶液,采用Inertsil C18色谱柱(250 mm×4.6 mm,5μm),以0.1%甲酸水为流动相A,甲醇为流动相B,进行梯度洗脱。检测波长:254 nm测定秦皮甲素、秦皮苷和秦皮乙素,340 nm测定秦皮素。进样量:10μL,柱温:30℃,流速:0.8 mL·min^-1,依次进样分析。结果秦皮甲素、秦皮苷、秦皮乙素和秦皮素质量浓度分别在78.60~1572、23.90~487.0、6.900~138.5、1.240~24.80 mg·L^-1内与峰面积呈良好的线性关系,相关系数分别为0.9996、0.9996、0.9995、0.9996;秦皮供试品中,四者的平均回收率分别为98.1%、98.9%、98.1%和98.5%;RSD分别为1.5%、1.6%、1.2%和0.90%(n=6)。结论建立的方法适用于研究秦皮药材中秦皮甲素、秦皮苷、秦皮乙素和秦皮素的含量测定。

Objective To establish a method for the determination of the contents of aesculin,aesculetin,fraxin and fraxetin in fraxini cortex by HPLC,and provide a quantitative method for quality evaluation of Fraxini Cortex.Methods Fraxini Cortex sample solution were prepared.The separation was performed on anInertsil C18(250 mm×4.6 mm,5μm)column,0.1%formic acid as mobile phase A,methanol as mobile phase B,gradient elution.Detection wavelengthwas set at 254 nm foraesculin,aesculetin and fraxin,340 nm for fraxetin.The injection volume was 10μL,the column temperature was 30℃,the flow rate was 0.8 mL·min-1.Results The calibration curve was linear within the range of 78.60-1572 mg·L-1(r=0.9996),23.90-487.0 mg·L^-1(r=0.9996),6.900-138.5 mg·L^-1(r=0.9995)and 1.240-24.80 mg·L-1(r=0.9996)for aesculin,aesculetin,fraxin and fraxetin,respectively.In Fraxini Cortex sample solution,the average recoveries were 98.1%(RSD=1.5%),98.9%(RSD=1.6%),98.1%(RSD=1.2%)and 98.5%(RSD=0.9%)for aesculin,aesculetin,fraxin and fraxetin,respectively.Conclusion The established method is suitable for study the constents of aesculin,aesculetin,fraxin and fraxetin in fraxini cortex.