HPLC法同时测定白芍、甘草药对提取物中9种有效成分

Simultaneous determination of nine constituents in combination extracts of Paeoniae Radix alba and Glycyrrhizae Radix et Rhizoma praeparata cum melle by HPLC

目的建立反向高效液相色谱法,同时测定白芍甘草药对提取物中芍药内酯苷、芍药苷、甘草苷、1,2,3,4,6-五没食子酰葡萄糖、芹糖基异甘草苷、异甘草苷、苯甲酰芍药苷、甘草酸、异甘草素9种成分。方法采用Dikma Technologies-C18(200mm×4.6mm,5μm),流动相为乙腈(A)-0.1%磷酸(B),梯度洗脱,体积流量1.0mL/min,检测波长为267nm(0～13min)、258 nm(13～17min)、230 nm(17～27min)、276 nm(27～42min)、360 nm(42～46 min)、276nm(46～50min)、230nm(50～53min)、275nm(53～55min)、250nm(55～90min),柱温30℃。结果在色谱条件下,芍药内酯苷、芍药苷、甘草苷、1,2,3,4,6-五没食子酰葡萄糖、芹糖基异甘草苷、异甘草苷、苯甲酰芍药苷、甘草酸、异甘草素的进样量分别在0.004 476～0.044 76μg,0.014 67～0.146 7μg,0.004 975～0.049 75μg,0.001 031～0.010 31μg,0.005 813～0.058 13μg,0.001 744～0.017 44μg,0.000 982～0.009 82μg,0.018 20～0.182 0μg,0.000 098～0.000 98μg范围内与色谱峰面积呈良好的线性关系;加样回收率(n=5)均在96.3%～103.4%,RSD均小于2.5%。结论本法快速、准确,重复性好,可更好地控制白芍炙甘草药对提取物的质量。

AIM To establish an HPLC method for simultaneously determining nine constituents （albiflorin, paeoniflorin, liquiritin, 1, 2, 3, 4, 6-pentgalloyl glucose, isoliquiritin apioside, isoliquiritin, benzoylpaeoniflorin, glycyrrhizic acid, isoliquiritigenin） in combination extracts of Paeoniae Radix alba and Glycyrrhizae Radix et Rhizoma cure melle. METHODS The separation was performed on a Dikma Technologies-C18 BDS （200 mm × 4. 6 mm, 5 μm） column. The mobile phase was composed of acetonitrile （A） -0. 1% phosphoric acid （B） in the gradient elution at the flow rate of 1.0 mL/min. Detection wavelengths were 267 nm （0-13 min） , 258 nm （ 13- 17min）, 230 nm （17-27min）, 276 nm （27-42 min）, 360 nm （42-46 min）, 276 nm （46-50 min）, 230 nm （50-53 min）, 275 nm （53-55min）, 250 nm （55-59 min）, 230nm （59-70 min）, 250 nm （70-95 min）, respectively. The column temperature was set at 30 ℃. RESULTS Albiflorin, peoniflorin, liquiritin, 1, 2, 3, 4, 6-pentgalloyl glucose, isoliquiritin apioside, isoliquiritin, benzoylpaeoniflorin, glycyrrhizic acid, isoliquiritigenin had good linearity in the ranges of 0.004 476-0. 044 76 μg, 0.014 67-0. 146 7 μg, 0. 004 975-0. 049 75μg, 0. 001 031-0. 010 31 μg, 0. 005 813-0. 058 13 μg, 0.001 744-0. 017 44 μg, 0. 000 982-0. 009 82 μg, 0. 018 20-0. 182 0 μg, 0. 000 098-0. 000 98 μg, respectively. The average recoveries of nine constituents were 96. 3% -103.4% with RSDs less than 2.5%. CONCLUSION The developed method is rapid, accurate with high reproducibility and stability, and is helpful to control the quality of the combination extracts of Paeoniae Radix alba and Glycyrrhizae Radix et Rhizoma cum melle.