RP-HPLC法同时测定加味五子衍宗方汤剂中3种活性成分的含量

RP-HPLC simultaneous determination of 3 active components in the traditional Chinese medicinal preparation modified Wuzi Yanzong prescription decoction

目的：建立RP-HPLC法同时测定加味五子衍宗方汤剂中绿原酸、金丝桃苷、淫羊藿苷的含量。方法：采用HypersilGOLD-C18（250 mm×4.6 mm,5μm）色谱柱,柱温25℃,以0.1%甲酸溶液（A）-含0.1%甲酸的甲醇（B）为流动相,线性梯度洗脱,流速1.0 mL.m in-1,程序检测波长为326 nm（0~17 m in,检测绿原酸）和350 nm（17.01~60 m in,检测金丝桃苷、淫羊藿苷）。结果：在58 m in内加味五子衍宗方汤剂中绿原酸、金丝桃苷、淫羊藿苷分离良好;依次在1.75~112μg.mL-1（r=0.9999）,1.69~108μg.mL-1（r=0.9999）,2.5~160μg.mL-1（r=0.9999）浓度范围内呈良好的线性关系;加样回收率（n=6）依次为101.9%,98.9%,99.3%。结论：本方法简便、可靠,重复性好,适用于测定加味五子衍宗方汤剂中绿原酸、金丝桃苷、淫羊藿苷的含量。

Objective：To develop an HPLC method for simultaneous determination of chlorogenic acid,hyperin and icariin in modified Wuzi Yanzong prescription decoction.Methods：The samples were separated by a Hypersil GOLD-C18（250 mm×4.6 mm,5 μm） column with a linear gradient elution system using 0.1%（v/v） formic acid solution（A） and methanol[containing 0.1%（v/v） formic acid]（B） as mobile phase at a flow rate of 1.0 mL·min-1 and the eluate was detected by programmed wavelength（0-17 min,326 nm for chlorogenic acid;17.01-60 min,350 nm for hyperin and icariin）.Results：Chlorogenic acid,hyperin and icariin were separated excellently in less than 58 min with the linear range of 1.75-112 μg·mL-1（r=0.9999）,1.69-108 μg·mL-1（r=0.9999）,2.5-160 μg·mL-1（r=0.9999）,with the recoveries（n=6） of 101.9%,98.9% and 99.3%,respectively.Conclusion：The established method is simple,and relible with good reproducibility for the quantitative analysis of active components of chlorogenic acid,hyperin and icariin in modified Wuzi Yanzong prescription decoction.