摘要:
Objective To establish a RP-HPLC method for simultaneously determining berberine hydrochloride, baicalin, emodin and chrysophanol in Sanhuang tablets. Methods The analysis was performed on an Thermo Hypersil GOLD aQ column (250 mm×4.6 mm, 5.0 μm) with gradient elution of methanol and 0.1%orthophosphoric acid at the flow rate of 1.0 ml/min. The column temperature was 30 , and the detection wave°Clength was 254 nm. Results The peaks of berberine hydrochloride, baicalin, emodin and chrysophanol were successfully separated from each other. The liner ranges of calibration curves were 0-0.6348 μg (r=0.9999), 0-1.0172 μg (r=0.9999), 0-0.8024 μg (r=0.9999), 0-0.9672 μg (r=0.9999). The average recoveries (n=9) were 101.03%, 99.81%, 99.35%, 99.54%, respectively. The limits of quantification (LOQs) were 0.0363, 0.0210, 0.0497, 0.0793 mg/g, respectively. The sample solution was stable within 24 h, the RSD (n=5) were 0.48%, 0.56%, 0.68%, 0.49%, respectively. The robustness was investigated by varying the conditions of column temperature (± 1 °C) and flow rate (± 0.1 ml/min), with the RSD s 0.57% and 0.42%, 0.44% and 0.65%, 0.74% and 0.29%, 0.46% and 0.56%, respectively. The contents of berberine hydrochloride, baicalin, emodin and chrysophanol in twelve samples were 4.6303-5.5866, 14.5514-18.8189, 0.1711-2.7984, 0.5299-2.9254 mg/tablet, respectively. Conclusion The establish method is validated to be suitable for quality control of Sanhuang tablets.%目的 建立同时测定三黄片中盐酸小檗碱、黄芩苷、大黄素、大黄酚含量的RP-HPLC法.方法 采用Thermo Hypersil GOLD aQ色谱柱(250 mm×4.6 mm,5.0μm),以甲醇-0.1%磷酸水溶液为流动相,进行梯度洗脱,流速1.0 ml/min,柱温30°C,检测波长254 nm.结果 盐酸小檗碱、黄芩苷、大黄素、大黄酚的分离度均符合要求,线性范围分别为0~0.6348μg(r=0.9999)、0~1.0172μg(r=0.9999)、0~0.8024μg(r=0.9999)、0~0.9672μg(r=0.9999);平均回收率(n=9)分别为101.03%、99.81%、99.35%、99.54%;定量限分别为0.0363、0.0210、0.0497、0.0793 mg/g;供试品溶液在24 h内稳定,RSD(n=5)分别为0.48%、0.56%、0.68%、0.49%;方法耐用性良好,在不同柱温(±1°C)条件下,RSD(n=3)分别为0.57%、0.42%、0.44%、0.65%,不同流速(±0.1 ml/min)条件下,RSD(n=3)分别为0.74%、0.29%、0.46%、0.56%.12批样品中盐酸小檗碱、黄芩苷、大黄素、大黄酚含量分别为4.6303~5.5866、14.5514~18.8189、0.1711~2.7984、0.5299~2.9254 mg/小片.结论 本法经方法学验证,适用于三黄片的质量控制.