HPLC-UV-ELSD同时测定知母黄柏药对提取物中盐酸小檗碱、黄柏碱、知母皂苷BⅡ

Quantitative Determination of Berberine Hydrochloride,Phellodendrine and Timosaponin BⅡ in Common Anemarrhena Rhizome and Amur Corktree Bark Extraction

目的:建立知母黄柏药对提取物中盐酸小檗碱、黄柏碱、知母皂苷BⅡ的HPLC-UV-ELSD含量测定方法。方法:采用HPLC-UV-ELSD方法,色谱柱:Dikma C18(4.6 mm×250 mm,5μm),流动相乙腈-0.2%冰醋酸水溶液,梯度洗脱,流速1.0mL·min-1,柱温30℃,UV检测波长280 nm,蒸发光散射检测器检测,漂移管温度46℃,氮气压力1.5 Bar。结果:盐酸小檗碱、黄柏碱及知母皂苷BⅡ的线性范围分别是1.008～3.528μg(r=0.999 5),0.322～1.127μg(r=0.999 6),0.702～2.457μg(r=0.999 8),平均加样回收率分别为100.11%,98.97%,99.56%;其RSD分别为1.08%,0.90%,1.37%。结论:该法简便快速,重复性良好,结果准确可靠,可用于知母黄柏药对提取物中主要有效成分的含量测定。

Objective：To establish a method for determination of berberine hydrochloride,phellodendrine and timosaponin B Ⅱ in common anemarrhena rhizome and amur corktree bark extraction.Method：HPLC-UV-ELSD was used in quantitative analysis with condition as follows：Dikma C18（4.6 mm × 250 mm,5 μm） was used in analysis;mobile phase was acetonitrile-0.2% acetic acid,gradient elution;flow rate was 1.0 mL.min-1;column temperature was at 30 ℃.Both UV and ELSD were used as detector.UV wavelength of 280 nm;evaporative light scattering detector drift tube temperature was at 46 ℃;nitrogen pressure：1.5 Bar.Result：The linear ranges of berberine hydrochloride,phellodendrine and timosaponin BⅡ were 1.008-3.528 μg（r=0.999 5）,0.322-1.127 μg（r=0.999 6）,0.702-2.457 μg（r=0.999 8）,the average recoveries were 100.11%,98.97%,99.56% with the RSD 1.08%,0.90%,1.37%.Conclusion：This quantitative determination method is simple,well reproducible,accurate and reliable.