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目的:建立双波长反相高效液相色谱(RP-HPLC)同时测定解毒祛瘀滋阴方阿魏酸、异阿魏酸和芍药苷的含量。方法:色谱柱为Dionex C18柱(4.6 mm×250 mm,5.0μm),流动相A乙腈,流动相B为0.1%甲酸溶液(体积比),梯度洗脱,流速为0.8 mL.min-1,检测波长:阿魏酸和异阿魏酸325 nm,芍药苷为275 nm,柱温25℃。结果:阿魏酸、异阿魏酸和芍药苷的质量浓度分别在0.15～15.0μg.mL-1(r=0.9999)、0.25～25.0μg.mL-1(r=0.9994)和1.50～60.0μg.mL-1(r=0.9999)内与峰面积呈良好的线性关系(n=5),平均回收率(n=9)分别为99.1%,98.3%,100.1%。结论:该法操作简单,灵敏度高,可用于解毒祛瘀滋阴方阿魏酸、异阿魏酸和芍药苷的同时测定。
Objective: To establish a dual-wavelength reversed-phase high performance liquid chromatography (RP-HPLC) for the simultaneous determination of ferulic acid, isoferulic acid and paeoniflorin in Jiedu Quyu Ziyin Recipe. METHODS: The column was Dionex C18 (4.6 mm × 250 mm, 5.0 μm). The mobile phase A was acetonitrile and the mobile phase B was 0.1% formic acid (volume ratio). The gradient elution was performed with a flow rate of 0.8 mL · min- Detection wavelength: ferulic acid and isoferulic acid 325 nm, paeoniflorin 275 nm, column temperature 25 ℃. Results: The concentrations of ferulic acid, isoferulic acid and paeoniflorin were 0.15-15.0μg.mL-1 (r = 0.9999), 0.25-25.0μg.mL-1 (r = 0.9994) and 1.50-60.0μg (n = 5). The average recoveries (n = 9) were 99.1%, 98.3% and 100.1%, respectively. Conclusion: The method is simple, high sensitivity, can be used for simultaneous determination of detoxification Quyu Ziyin Fang ferulic acid, isoferulic acid and paeoniflorin.