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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 establish a RP-HPLC method for simultaneous determination of chlorogenic acid, hyperoside and icariin in Jiawei Wuzi Yanzong Decoction. METHODS: A HypersilGOLD-C18 (250 mm × 4.6 mm, 5 μm) column was used. The column temperature was 25 ℃. The mobile phase consisted of 0.1% formic acid solution (A) The flow rate was 1.0 mL.m in-1, and the detection wavelength was 326 nm (0-17 m in, chlorogenic acid detection) and 350 nm (17.01-60 nm). The detection of hyperoside and icariin was carried out. Results: The contents of chlorogenic acid, hyperoside and icariin in Wuzi Yanzong Decoction were well separated within 58 min, followed by 1.75 ~ 112μg.mL-1 (r = 0.9999), 1.69-108μg.mL (R = 0.9999) and 2.5 ~ 160μg.mL-1 (r = 0.9999). The recoveries were nominally 101.9%, 98.9% and 99.3%, respectively. Conclusion: The method is simple, reliable and reproducible. It is suitable for the determination of chlorogenic acid, hyperoside and icariin in Jiawei Wuzi Yanzong Decoction.