高效液相色谱法测定小儿咽扁颗粒中胆红素含量

Content Determination of Bilirubin in Xiao′er Yanbian Granules by HPLC

目的 建立测定小儿咽扁颗粒中胆红素含量的高效液相色谱(HPLC)法。方法 色谱柱为Scientific Acclaim^(TM)120 C_(18)柱(250 mm×4.6 mm,5μm),流动相为甲醇-三氯甲烷-0.1%磷酸溶液(88∶6∶6,V/V/V),流速为1.0 mL/min,检测波长为450 nm,柱温为30℃,进样量为10μL。结果 胆红素进样量在33.47~133.89 ng范围内与峰面积积分值线性关系良好(r=1.000);精密度、稳定性、重复性试验结果的RSD均小于6%;平均提取回收率为98.77%,RSD为7.79%(n=9)。结论 该方法操作简便,结果准确,专属性强,可用于制剂中体外培育牛黄的含量测定。

ObjectiveTo establish a high-performance liquid chromatography (HPLC) method for the content determination of bilirubin in Xiao′er Yanbian Granules.MethodsThe chromatographic column was Scientific Acclaim^(TM)120 C_(18)column (250 mm×4.6 mm,5μm),the mobile phase was methanol-trichloromethane-0.1%phosphoric acid solution (88∶6∶6,V/V/V),the flow rate was 1.0 mL/min,the detection wavelength was 450 nm,the column temperature was 30℃,and the injection volume was 10μL.ResultsThe linear range of bilirubin was 33.47-133.89 ng(r=1.000).The RSDs of precision,stability and repeatability tests were lower than 6%.The average recovery of bilirubin was 98.77%with an RSD of 7.79%(n=9).ConclusionThe method is simple,accurate and specific,which can be used for the content determaination of in-vitro-cultured calculus bovis (ICCB) in the preparation.