HPLC-ELSD法测定胃康颗粒中人参皂苷Rb1和黄芪甲苷的含量

Detectione of the contents of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules by HPLC-ELSD

目的优化胃康颗粒中人参皂苷Rb1和黄芪甲苷的提取方法,并建立其含量测定方法。方法以人参皂苷Rb1和黄芪甲苷的含量作为指标,采用单因素考察法对提取工艺进行优化;采用高效液相色谱-蒸发光散射法(HPLC-ELSD),XBridge?Shield RP18(4.6 mm×250 mm,5μm)为色谱柱;乙腈-水(32:68)为流动相;柱温为30℃;漂移管温度为60℃,载气流量为1.7 SLM,建立测定胃康颗粒中人参皂苷Rb1和黄芪甲苷含量的方法。结果当采用甲醇回流提取1.5 h,正丁醇提取5次,氨水洗涤2次时,人参皂苷Rb1和黄芪甲苷的提取含量较高;建立的HPLC-ELSD法测定人参皂苷Rb1和黄芪甲苷含量,线性关系良好(r> 0.999 7),日内日间精密度均小于1%,加样回收率分别为95.65%和100.57%,稳定性和重复性的RSD均小于3%,含量分别为2.863 0 mg/g和0.257 6 mg/g,RSD分别为0.62%和1.51%。结论优化了胃康颗粒中人参皂苷Rb1和黄芪甲苷的提取方法,建立了可靠、准确、重现性好的测定胃康颗粒中人参皂苷Rb1和黄芪甲苷含量的HPLCELSD方法。

Objective To optimize the extraction method and develop the detection method of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules. Methods The extraction process of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules were optimized by single factor investigation, with the contents of ginsenoside Rb1 and astragaloside Ⅳ as optimization indicators. The HPLC-ELSD method was developed for the detection of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules.Separation was carried out on an XBridge?Shield RP18 column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-water(32:68)at the flow rate of 1 ml/min. The column temperature was maintained at 30 ℃. The drift tube temperature was set at 60 ℃, and the carrier gas flow rate was 1.7 SLM. Results The optimized extraction methods of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules were as the following: methanol reflux extraction for 1.5 h, and n-butanol extraction and ammonia washed for 5 and 2 times, respectively. The HPLC-ELSD method was established to detect the contents of ginsenoside Rb1 and astragaloside Ⅳ. The linear relationship was good(r > 0.999 7). The intra-day and inter-day precision was less than 1%.The recovery rates were 95.65% and 100.57%. The stability and repeatability RSD were less than 3%. The contents were2.863 0 mg/g and 0.257 6 mg/g. The RSDs were 0.62% and 1.51%, respectively. Conclusion The extraction method of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules is optimized, and a reliable, accurate and reproducible HPLC-ELSD method for the detection of the contents of ginsenoside Rb1 and astragaloside Ⅳ in Weikang granules is established.