HPLC-DAD法同时测定柴黄片中6种成分的含量

目的:建立同时测定柴黄片中6种成分黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、柴胡皂苷a和柴胡皂苷d含量的方法。方法:采用高效液相色谱-二极管阵列检测器(HPLC-DAD)法检测同一厂家的3批柴黄片。色谱柱为Agilent Eclipse XDB-C18;流动相为乙腈-磷酸三乙胺水溶液(pH调至7.0),梯度洗脱;流速为1.0mL/min;检测波长为210nm(柴胡皂苷a、柴胡皂苷d)和277nm(黄芩苷、汉黄芩苷、黄芩素、汉黄芩素);柱温为30℃;进样量为5μL。结果:黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、柴胡皂苷a和柴胡皂苷d检测进样量线性范围分别为0.3795~7.5904、0.08296~1.6592、0.03939~0.7878、0.04072~0.8144、0.04045~0.8090、0.03863~0.7726μg(r均≥0.9993),检测限分别为0.008、0.007、0.005、0.005、0.020、0.018μg/mL,定量限分别为0.025、0.022、0.015、0.015、0.060、0.054μg/mL,精密度、重复性和稳定性试验(48h)中的RSD<1.5%(n=6),平均加样回收率分别为98.46%、97.06%、100.90%、96.13%、96.91%、96.57%(RSD<2.0%,n=6)。结论:建立的同时测定柴黄片中6种成分含量的方法操作简便、结果准确、重复性好,可用于该制剂的质量控制。

Simultaneous Determination of 6 Components in Chaihuang Tablets by HPLC-DAD

OBJECTIVE:To establish a method for simultaneous determination of 6 components in Chaihuang tablets,such as baicalin,wogonoside,baicalein,wogonin,saikosaponin a and saikosaponin d in Chaihuang tablets. METHODS:HPLC-DAD method was used to detect 3 batches of Chaihuang tablets from same manufacturers. The determination was performed on Agilent Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-triethylamine phosphate aqueous solution(pH adjusted to 7.0, gradient elution)at flow rate of 1.0 mL/min. The detection wavelengths were set at 210 nm(saikosaponin a,saikosaponin d)and 277 nm(baicalin,wogonoside,baicalein,wogonin). The column temperature was 30 ℃,and sample size was 5 μL. RESULTS: The linear ranges of baicalin,wogonoside,baicalein,wogonin,saikosaponin a and saikosaponin d were 0.379 5-7.590 4 μg, 0.082 96-1.659 2 μg,0.039 39-0.787 8 μg,0.040 72-0.814 4 μg,0.040 45-0.809 0 μg,0.038 63-0.772 6 μg(all r≥0.999 3), respectively. The limits of detection were 0.008,0.007,0.005,0.005,0.020 and 0.018 μg/mL. The limits of quantitation were 0.025,0.022,0.015,0.015,0.060,0.054 μg/mL. RSDs of precision,reproducibility and stability tests(48 h)were all lower than 1.5%(n=6). Average recoveries were 98.46%,97.06%,100.90%,96.13%,96.91%,96.57%(RSD<2.0%,n=6). CONCLUSIONS:Established method is simple,accurate and reproducible for 6 components in Chaihuang tablets,and can be used for quality control of the tablet.