高效液相色谱法测定黄连上清片中3种生物碱

Determination of three kinds of alkaloids in Huanglian Shangqing Tablet

目的建立黄连上清片(黄柏、黄连、大黄、连翘、薄荷、黄芩等)中盐酸小檗碱、盐酸巴马汀和盐酸药根碱的测定方法。方法采用甲醇超声提取,反相离子对高效液相色谱法测定。C18色谱柱(250 mm×4.6 mm,5μm);柱温40℃;流动相为0.01 mol/L庚烷磺酸钠与0.01 mol/L磷酸二氢钾等量混合(用磷酸调pH为3.1)-乙腈(70∶30);体积流量为1.0mL/min;检测波长为345 nm,进样量为10μL。结果 3种生物碱成分色谱分离行为良好,盐酸小檗碱、盐酸巴马汀和盐酸药根碱的线性范围分别为10.671 1～64.026 6μg(r=0.999 99)、1.123 2～6.739 2μg(r=0.999 16)和1.198 8～7.192 8μg(r=0.999 66);平均回收率分别为99.66%、98.25%和97.40%,RSD分别为1.2%、0.6%和0.8%(n=6)。结论该方法简单易行,专属性强,准确性好,重复性好,可用于黄连上清片的质量控制。

AIM To develop a method for determining berberine hydrochloride,palmatine hydrochloride and jatrorrhizine hydrochloride in Huanglian Shangqing Tablet（Phellodendri chinensis Cortex,Coptidis Rhizoma,Rhei Radix et Rhizoma,Forsythiae Fructus,Menthae Herba,Scutellariae Radix,etc.）.METHODS The samples were extracted by ultrasonication with methanol,and an ion pair RP-HPLC method was applied to determining three kinds of alkaloids.The column of C18（250 mm×4.6 mm,5 μm） at the temperature of 40 ℃ was used with the mobile phase consisting of the mixed solution of 0.01 mol/L sodium-heptanesulfonate solution and equal volume of 0.01 mol/L potassium dihydrogen phosphate solution（adjusting pH to 3.1 with phosphoric acid） combined with acetonitrile（70∶30）.The flow rate was 1.0 mL/min,and the detection wavelength was set at 345 nm,and the sample size was 10 μL.RESULTS Three kinds of alkaloids were seperated perfectly and good linear relationships were obtained in the following ranges,10.671 1-64.026 6 μg（r=0.999 99） for berberine,1.123 2-6.739 2 μg（r=0.999 16） for palmatine,and 1.198 8-7.192 8 μg（r=0.999 66） for jatrorrhizine.The recoveries were 99.66%,98.25% and 97.40%,respectively,and RSDs were 1.2%,0.6% and 0.8%（n=6）,respectively.CONCLUSION The experiment shows that the developed method is simple,specific,accurate and reproducible,and can be used for the quality control of Huanglian Shangqing Tablet.