高效液相色谱法同时测定启脾丸中9种成分含量

Simultaneous Determination of Nine Components in Qipi Pills by HPLC

目的 建立同时测定启脾丸中9种成分含量的高效液相色谱-二极管阵列检测器(HPLC-DAD)法。方法 色谱柱为Agilent Extend C_(18)柱(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈(1∶1,V/V)-0.08%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长分别为237 nm(甘草苷、甘草酸铵)、283 nm(橙皮苷)、203 nm(人参皂苷Rg_1、人参皂苷Re)、220 nm(白术内酯Ⅲ、白术内酯Ⅱ、白术内酯Ⅰ、茯苓酸),柱温为30℃,进样量为10μL。结果 甘草苷、甘草酸铵、橙皮苷、人参皂苷Rg_1、人参皂苷Re、白术内酯Ⅲ、白术内酯Ⅱ、白术内酯Ⅰ、茯苓酸的质量浓度分别在3.66~58.48μg/mL、6.21~99.41μg/mL、9.53~152.54μg/mL、3.06~48.89μg/mL、1.51~24.11μg/mL、2.54~40.60μg/mL、1.24~19.78μg/mL、2.10~33.64μg/mL、3.57~57.08μg/mL范围内与峰面积线性关系良好(r≥0.999 5,n=6);精密度、重复性、稳定性试验结果的RSD均小于2.0%(n=6);平均加样回收率分别为100.34%,101.35%,101.18%,99.65%,98.73%,98.86%,100.86%,99.30%,100.86%,RSD分别为1.43%,1.52%,0.68%,1.43%,1.00%,1.19%,1.31%,0.75%,1.36%(n=6)。样品中9种成分的含量分别为0.526 4~1.100 7 mg/g、0.956 2~1.700 2 mg/g、1.125 2~1.876 2 mg/g、0.570 9~0.825 6 mg/g、0.101 4~0.502 6 mg/g、0.226 9~0.611 7 mg/g、0.099 4~0.370 5 mg/g、0.152 5~0.416 4 mg/g、0.395 2~0.755 6 mg/g(n=3)。结论 该方法操作简便、专属性强、稳定性好、结果准确,可用于启脾丸中9种成分含量的同时测定。

Objective To establish a high-performance liquid chromatography-diode array detector(HPLC-DAD)method for simultaneous determination of nine components in Qipi Pills.Methods The chromatographic column was Agilent Extend C18 column(250 mm×4.6 mm,5μm),the mobile phase was methanol acetonitrile(1∶1,V/V)-0.08%phosphoric acid aqueous solution(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was set at 237 nm for liquiritin and ammonium glycyrrhizinate,283 nm for hesperidin,203 nm for ginsenoside Rg1 and ginsenoside Re,220 nm for atracylenolideⅢ,atracylenolideⅡ,atracylenolideⅠ,and pachymic acid,the column temperature was 30℃,and the injection volume was 10μL.Result The linear ranges of liquiritin,ammonium glycyrrhizinate,hesperidin,ginsenoside Rg1,ginsenoside Re,atracylenolideⅢ,atracylenolideⅡ,atracylenolideⅠand pachymic acid were 3.66-58.48μg/mL,6.21-99.41μg/mL,9.53-152.54μg/mL,3.06-48.89μg/mL,1.51-24.11μg/mL,2.54-40.60μg/mL,1.24-19.78μg/mL,2.10-33.64μg/mL,and 3.57-57.08μg/mL(r≥0.9995,n=6).The RSDs of precision,repeatability,and stability test results were all lower than 2.0%(n=6).The average recoveries of the above nine components were 100.34%,101.35%,101.18%,99.65%,98.73%,98.86%,100.86%,99.30%and 100.86%,with RSDs of 1.43%,1.52%,0.68%,1.43%,1.00%,1.19%,1.31%,0.75%,and 1.36%(n=6),respectively.The contents of the above nine components in samples were in the ranges of 0.5264-1.1007 mg/g,0.9562-1.7002 mg/g,1.1252-1.8762 mg/g,0.5709-0.8256 mg/g,0.1014-0.5026 mg/g,0.2269-0.6117 mg/g,0.0994-0.3705 mg/g,0.1525-0.4164 mg/g,and 0.3952-0.7556 mg/g(n=3),respectively.Conclusion The method is simple,has strong specificity,good stability,and accurate results,which can be used for the simultaneous determination of nine components in Qipi Pills.