目的 建立HPLC同时测定康复春口服液中毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪甲苷和人参皂苷Rb₁的方法。方法 采用Diamonsil C₁₈色谱柱（250 mm×4.6 mm，5 μm）；流动相：乙腈–0.2%甲酸溶液，梯度洗脱；检测波长254 nm（0～30 min，检测毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮和芒柄花素）、210 nm（30～36 min，检测黄芪甲苷）和203 nm（36～50 min，检测人参皂苷Rb₁）；体积流量：0.8 mL/min；柱温：30℃；进样量为20 μL。结果 毛蕊异黄酮葡萄糖苷、芒柄花苷、毛蕊异黄酮、芒柄花素、黄芪甲苷、人参皂苷Rb₁分别在3.29～82.25μg/mL（r＝0.999 1）、4.77～119.25μg/mL（r＝0.999 8）、4.16～104.00μg/mL（r＝0.999 2）、11.33～283.25μg/mL（r＝0.999 7）、7.39～184.75μg/mL（r＝0.999 9）、2.05～51.25μg/mL（r＝0.999 5）线性关系良好；平均加样回收率分别为97.98%、98.45%、97.38%、99.72%、98.67%、96.99%，RSD值分别0.91%、1.41%、0.78%、1.09%、1.22%、0.85%。结论 方法专属性强，结果准确，重复性好，为更好地评价康复春口服液的质量提供参考。

Objective To develop an HPLC method for simultaneous determination of calycosin7-O-β-D-glucopyranoside, ononin, calycosin, formononetin, astragaloside IV, and ginsenoside Rb₁ in Kangfuchun Oral Liquid. Methods The determination was carried out on Diamonsil C₁₈ chromatographic column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.2% formic acid solution with gradient elution. The detection wavelengths were set at 254 nm in 0-30 min to determine calycosin7-O-β-D-glucopyranoside, ononin, calycosin, and formononetin, 210 nm in 30-36 min to determine astragaloside IV, and 203 nm in 36-50 min to determine ginsenoside Rb₁. The flow rate was 0.8 mL/min, temperature of column was set at 30℃, and volume of injection was 20 μL. Results The linear range of calycosin7-O-β-D-glucopyranoside, ononin, calycosin, formononetin, astragaloside IV, and ginsenoside Rb₁ were 3.29-82.25 μg/mL(r=0.999 1), 4.77-119.25 μg/mL (r=0.999 8), 4.16-104.00 μg/mL(r=0.999 2), 11.33-283.25 μg/mL (r=0.999 7), 7.39 -184.75 μg/mL (r=0.999 9), 2.05 -51.25 μg/mL (r=0.999 5), respectively. The average recoveries were 97.98%), 98.45% 97.38%, 99.72%, 98.67%, and 96.99%, respectively. The corresponding RSD values were 0.91%, 1.41%, 0.78%, 1.09%, 1.22%, and 0.85%, respectively. Conclusion The method has strong specificity, accurate results, and good reproducibility which can provide a reference for quality of Kangfuchun Oral Liquid.

河南省医学科技攻关计划项目（201403190）