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.