摘要
目的 建立同时测定五味香连丸中盐酸小檗碱、黄连碱、巴马汀、延胡索乙素、木香烃内酯、去氢木香内酯、吴茱萸碱、吴茱萸次碱、柠檬苦素、芍药苷 10 种成分含量的反相高效液相色谱法.方法 色谱柱为Waters SunFire C18 柱(250 mm×4.6 mm,5 μm),流动相为含 0.02%三乙胺的 0.02 mol/L磷酸二氢钾溶液(用磷酸调pH至 3.5)-乙腈(梯度洗脱),流速为 1.0 mL/min,柱温为 35℃,检测波长为 225 nm,进样量为 10 μL.结果 盐酸小檗碱、黄连碱、巴马汀、延胡索乙素、木香烃内酯、去氢木香内酯、吴茱萸碱、吴茱萸次碱、柠檬苦素、芍药苷的质量浓度分别在 6.852～51.39 μg/mL、2.956～22.17 μg/mL、2.918～21.88 μg/mL、4.643～34.82 μg/mL、4.871～36.53 μg/mL、4.007～30.05 μg/mL、4.519～33.89 μg/mL、4.482～33.62 μg/mL、2.724～20.43 μg/mL、2.809～21.07 μg/mL范围内与峰面积线性关系良好(r≥0.999 2,n = 6);定量限分别为 0.66,0.31,0.29,0.45,0.48,0.41,0.42,0.46,0.27,0.28 μg/mL;精密度、稳定性、重复性试验结果的RSD均小于 2.0%(n = 6);回收率分别为100.36%,98.74%,98.71%,99.67%,99.86%,99.59%,98.70%,98.38%,98.25%,98.72%,RSD 分别为0.89%,1.02%,1.04%,1.17%,0.84%,1.15%,1.33%,1.20%,1.07%,1.23%(n = 6).3 批样品中上述成分的平均含量分别为 1.165～1.192 mg/g、0.523 1～0.527 6 mg/g、0.499 7～0.501 1 mg/g、0.681 4～0.688 9 mg/g、0.697 7～0.700 2 mg/g、0.606 8～0.613 4 mg/g、0.580 1～0.583 7 mg/g、0.427 7～0.442 2 mg/g、0.410 1～0.416 5 mg/g、0.400 2～0.401 3 mg/g(n = 2).结论 所建立的方法简便可行、结果准确可靠、稳定性好,可用于同时测定五味香连丸中 10 种成分的含量.
Abstract
Objective To establish a reverse-phase high-performance liquid chromatography(RP-HPLC)method for the simultaneous determination of 10 components in Wuwei Xianglian Pills,including berberine ydrochloride,coptisine,palmatine,tetrahydropalmatine,costunolide,dehydrocostuslactone,evodiamine,rutaecarpine,limonin,and paeoniflorin.Methods The chromatography column was Waters SunFire C18 column(250 mm×4 6 mm,5 μm),the mobile phase was 0.02 mol/L potassium dihydrogen phosphate solution containing 0.02％triethylamine(the pH was ajusted to 3.5 with phophoric acid)-acetonitrile(gradient elution),the flow rate was 1.0 mL/min,the column temperature was 35℃,the detection wavelength was 225 nm,and the injection volume was 10 μL.Results The linear ranges of berberine hydrochloride,coptisine,palmatine,tetrahydropalmatine,costunolide,dehydrocostuslactone,evodiamine,rutaecarpine,limonin and paeoniflorin were 6.852-51.39 μg/mL,2.956-22.17 μg/mL,2.918-21.88 μg/mL,4.643-34.82 μg/mL,4.871-36.53 μg/mL,4.007-30.05 μg/mL,4.519-33.89 μg/mL,4.482-33.62 μg/mL,2.724-20.43 μg/mL,and 2.809-21.07 μg/mL(r≥0.999 2,n = 6),respectively.The limits of quantification(LOQ)were 0.66,0.31,0.29,0.45,0.48,0.41,0.42,0.46,0.27,0.28 μg/mL,respectively.The RSDs of precision,stability,and repeatability test results were all lower than 2.0％(n = 6).The average recoveries of the above 10 components were 100.36％,98.74％,98.71％,99.67％,99.86％,99.59％,98.70％,98.38％,98.25％,98.72％,with RSDs of 0.89％,1.02％,1.04％,1.17％,0.84％,1.15％,1.33％,1.20％,1.07％,and 1.23％(n = 6),respectively.The average contents of the above 10 components in three batches of samples were 1.165-1.192 mg/g,0.523 1-0.527 6 mg/g,0.499 7-0.501 1 mg/g,0.681 4-0.688 9 mg/g,0.697 7-0.700 2 mg/g,0.606 8-0.613 4 mg/g,0.580 1-0.583 7 mg/g,0.427 7-0.442 2 mg/g,0.410 1-0.416 5 mg/g,and 0.400 2-0.401 3 mg/g(n = 2),respectively.Conclusion The established method is convenient and feasible,with accurate and reliable results and good stability,which can be used for the simultaneous determination of 10 components in Wuwei Xianglian Pills.
维普
万方数据