全自动在线固相萃取-高效液相色谱法快速测定中药复方中小檗碱的含量

目的:建立在线固相萃取-HPLC方法测定中药复方中小檗碱的含量.方法:通过双梯度高效液相系统中的一个泵(上样泵)实现净化,再经过阀切换将目标物从固相萃取柱中切换至分析柱中进行测定.上样泵以乙腈-500 mmol· L-1醋酸铵溶液为流动相,采用硅胶基质强阳离子交换小柱(4.6 mm×50 mm,300(A))为在线固相纯化柱,采用梯度洗脱方式进行样品净化；分析泵以乙腈-30 mmo1·L-1磷酸二氢钾为流动相,采用Acclaim C18(4.6 mm×250 mm,5μm)柱,柱温35℃,流速1.0mL· min -,UV检测波长为345 nm.结果:小檗碱在1.24 ～ 124 mg·L-1线性关系较好(r =0.999),平均回收率99.5％.结论:方法快速、简便,专属性、重复性较好,测定结果准确.

Automated Online-SPE-HPLC for Rapid Determination of the Content of Berberine in the Chinese Traditional Medicine Compound

Objective:To establish the method for determining the content of berberine in traditional Chinese medicine with online solid phase extraction-HPLC.Method:Using one of the dual gradient pumps to enrich and purify the sample solution,then wash the aimed analyte into analytical column by switching the valve to another position after removing of matrix interference substance.Silica-based strong cation exchange column(300 ,4.6 mm×50 mm) was used as online-SPE column with a mobile phase consisting of acetonitrile-500 mmol·L-1 ammonium acetate in a gradient elution.Acclaim C18(4.6 mm×250 mm,5 μm) was used as analytical column with a mobile phase of acetonitrile-30 mmol·L-1 KH2PO4 at a flow rate of 1.0 mL · min-1.UV detection wavelength was set up at 345 nm.Result:There is a good linear relationship(r=0.999) within the concentration range of 1.24-124 mg · L-1.The average recovery was 99.5%.Conclusion:This method was simple,fast and has a better speciality.The results of determination were reliable and reproducible by validating for accuracy,precision and reproducibility.