HPLC切换波长法同时测定健脾止泻宁颗粒中盐酸小檗碱和黄芩苷的含量

摘要

目的:建立同时测定健脾止泻宁颗粒中盐酸小檗碱和黄芩苷含量的方法.方法:采用高效液相色谱切换波长法.色谱柱为Hypersil BDS C18,流动相为甲醇-0.45％磷酸溶液-三乙胺(50:49:1,V/V/V),流速为1.0 mL/min,检测波长为265 nm(盐酸小檗碱)、280 nm(黄芩苷),柱温为30℃,进样量为10μL.结果:盐酸小檗碱和黄芩苷检测进样量线性范围分别为60.3～312.8 ng(r=0.9997)、81.5～368.9 ng(r=0.9999);定量限分别为0.6668、0.7740 ng,检测限分别为0.2226、0.2580 ng;中间精密度、稳定性、重复性试验的RSD均小于1.0％;加样回收率分别为96.48％～99.30％(RSD=1.06％,n=6)、95.20％～99.39％(RSD=1.66％,n=6);耐用性试验的RSD均小于2.0％.结论:该方法操作简便,精密度、稳定性、重复性、准确度、耐用性均较好,可用于健脾止泻宁颗粒中盐酸小檗碱和黄芩苷含量的同时测定.%OBJECTIVE:To establish the method for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules. METHODS:HPLC switching walvelength method was adopted. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-0.45％ phosphoric acid solution-triethylamine(50:49:1,V/V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 265 nm(berberine hydrochloride)and 280 nm(baicalin). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride and baicalin were 60.3-312.8 ng(r=0.9997)and 81.5-368.9 ng(r=0.9999). The limits of quantitation were 0.6668,0.7740 ng,andthe limits of detection were 0.2226,0.2580 ng,respectively. RSDs of intermediate precision,stability and repeatability tests were all lower than 1.0％. The recoveries were 96.48％-99.30％(RSD=1.06％,n=6) and 95.20％-99.39％(RSD=1.66％,n=6), respectively. RSDs of durability test were all lower than 2.0％. CONCLUSIONS:The method is simple, precise, stable, reproducible,accurate and durable. It can be used for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules.