HPLC法同时测定白蒲黄片中5种成分的含量

Simultaneous Determination of 5 Components in Baipuhuang Tablets by HPLC

目的:建立同时测定白蒲黄片中白头翁皂苷B_4、咖啡酸、黄芩苷、盐酸巴马汀和盐酸小檗碱含量的方法。方法:采用高效液相色谱法。色谱柱为Agela Technologies Venusil XBP C_(18)(L),流动相为乙腈-0.05 mol/L磷酸二氢钾溶液(梯度洗脱),流速为0.8mL/min,检测波长为203 nm(白头翁皂苷B_4)和323 nm(咖啡酸、黄芩苷、盐酸巴马汀和盐酸小檗碱),柱温为30℃,进样量为10μL。结果:白头翁皂苷B_4、咖啡酸、黄芩苷、盐酸巴马汀和盐酸小檗碱检测进样量线性范围分别为0.081 41~8.141μg(r=0.999 8)、0.018 71~1.871μg(r=0.999 4)、0.037 33~3.733μg(r=0.999 2)、0.028 85~2.885μg(r=0.999 6)、0.027 58~2.758μg(r=0.999 7);定量限分别为0.009、0.006、0.008、0.011、0.013 ng,检测限分别为0.030、0.020、0.025、0.034、0.036 ng;精密度、稳定性、重复性试验的RSD<2.0%;加样回收率分别为97.39%~102.34%(RSD=1.81%,n=6)、96.77%~98.92%(RSD=0.85%,n=6)、97.38%~103.72%(RSD=2.46%,n=6)、96.73%~102.01%(RSD=2.22%,n=6)、96.47%~101.07%(RSD=1.61%,n=6)。结论:该方法操作简便、结果准确、重复性好,可用于白蒲黄片中白头翁皂苷B_4、咖啡酸、黄芩苷、盐酸巴马汀和盐酸小檗碱含量的同时测定。

OBJECTIVE： To establish a method for simultaneous determination of pulchinenoside B4, caffeic acid, baicalin, palmatine hydrochloride and berberine hydrochloride in Baipuhuang tablets. METHODS： HPLC method was adopted. The determination of Agela Technologies Venusil XBP C18（L） with mobile phase consisted of acetonitrile-0.05 mol/L monopotassium phosphate （gradient elution） at the flow rate of 0.8 mL/min. The detection wavelengths were set at 203 nm （pulchinenoside B4） and 323 nm （caffeic acid, baicalin, palmatine hydrochloride, berberine hydrochloride）. The column temperature was set at 30 ℃, and the sample size was 10 μL. RESULTS： The linear ranges were 0.081 41-8.141μg for pulchinenoside B4（r=0.999 8）, 0.018 71-1.871 μg for caffeic acid （r=0.999 4）, 0.037 33-3.733 μg for baicalin （r=0.999 2）, 0.028 85-2.885 μg for palmatine hydrochloride （r=0.999 6） and 0.027 58-2.758 μg for berberine hydrochloride （r=0.999 7）. The limits of quantitation were 0.009, 0.006, 0.008, 0.011, 0.013 ng, respectively. The limits of detection were 0.030, 0.020, 0.025, 0.034, 0.036 ng, respectively. RSDs of precision, stability and reproducibility tests were all lower than 2.0%. The average recoveries were 97.39%-102.34%（RSD=1.81%,n=6）, 96.77%-98.92%（RSD=0.85%,n=6）, 97.38%-103.72%（RSD=2.46%,n=6）, 96.73%-102.01%（RSD=2.22%,n=6）and 96.47%-101.07%（RSD=1.61%,n=6）, respectively. CONCLUSIONS： The method is simple, accurate and reproducibility, and can be used for simultaneous determination of pulchinenoside B4, caffeic acid, baicalin, palmatine hydrochloride and berberine hydrochloride in Baipuhuang tablet.