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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 HPLC-UV-ELSD method for the determination of berberine hydrochloride, cortex phellodendri, and timosaponin B Ⅱ in the extracts of Anemarrhena asphodeloides. Methods: The HPLC-UV-ELSD method was used. The chromatographic column was Dikma C18 (4.6 mm × 250 mm, 5 μm). The mobile phase was acetonitrile-0.2% glacial acetic acid and eluted with a gradient of 1.0 mL · min- ℃, UV detection wavelength 280 nm, evaporative light scattering detector detection, drift tube temperature 46 ℃, nitrogen pressure 1.5 Bar. Results: The linear ranges of berberine hydrochloride, cypermethrin and timosaponin B Ⅱ were 1.008-3.528μg (r = 0.999 5), 0.322-1.27μg (r = 0.999 6), 0.702-2.457μg ). The average recoveries were 100.11%, 98.97% and 99.56%, respectively. The RSDs were 1.08%, 0.90% and 1.37% respectively. Conclusion: The method is simple and rapid with good reproducibility and accurate and reliable results. It can be used for the determination of the main active ingredients in the extracts of Anemarrhena asperribate.