SPE-HPLC法检测苍耳子复方制剂中苍术苷的含量

目的:建立苍耳子复方制剂中毒性成分苍术苷的含量测定方法。方法:应用正、反相硅胶柱层析分离方法自制苍术苷对照品;复方样品经大孔树脂柱和固相萃取小柱前处理,再应用离子对色谱测定苍术苷的含量。采用Agilent Eclipse XDB-C₁₈(250 mm×4.6 mm,5.0 μm)色谱柱,以乙腈-0.15%磷酸二氢钠溶液(含0.12% 四丁基氢氧化铵,磷酸调pH至3.5)(39:61)为流动相,流速1.0 mL·min^-1,柱温30 ℃,紫外检测波长203 nm。结果:苍术苷对照品经HPLC归一化法分析纯度大于98.0%;苍术苷进样量在0.10~2.59 μg范围内线性关系良好(r=1.000),平均回收率为95.7%(RSD=1.2%,n=6)。测定不同产地、不同剂型样品5批,苍术苷含量分别为226.3、464.6、386.1、66.0、1 044.7 μg·g^-1。结论:所建立的方法经方法学验证,可用于苍耳子复方制剂中毒性成分苍术苷的质量控制。

Objective:To establish a method for determination of atractyloside,a toxic component,in compound preparations containing Xanthii Fructus.Methods:Atractyloside reference was isolated and purified by normal,reversed phase silica gel column chromatography.Macroporous resin and solid phase extraction were used for sample pre-treatment,the atractyloside was detected with ion-pair chromatography.The chromatographic separation was carried out with an Agilent Eclipse XDB-C₁₈ column(250 mm×4.6 mm,5.0 μm);the mobile phase was composed of acetonitrile-0.12% tetrabutylammonium hydroxide in 0.15% sodium dihydrogen phosphate solution(pH 3.5 adjusted with phosphoric acid)at the proportin of 39:61,the flow rate of 1.0 mL·min^-1,the column temperature was 30 ℃,and the detection wavelength was 203 nm.Results:The purity of atractyloside reference substance was detected by HPLC,which was over 98.0%.Atractyloside had good linear relationship in the range of 0.10-2.59 μg and the correlation coefficient was 1.000,and the average recovery was 95.7% with the RSD being 1.2%(n=6);the content results of 5 samples from different origins and varies of formulations were 226.3,464.6,386.1,66.0 and 1 044.7 μg·g^-1.Conclusion:According to methodology validation,the developed method can be used for quality control of compound preparations containing Xanthii Fructus.