摘要
In this study, a novel and simple high performance liquid chromatography with diode array detection （HPLC-DAD） method for the simultaneous qualitative and quantitative determination of 12 bioactive components in Gegen Qinlian pill was developed. The separation was performed on a Kromasil C18 column （250 mm×4.6 ram, 5.0 μm） by gradient elution with acetonitrile and 0.02 mol/L ammonium acetate （containing 0.3% triethylamine, and adjusted pH to 4.3 using 1% glacial acetic acid） as the mobile phase at a flow rate of 0.7 mL/min. Three different detection wavelengths （250, 280, 346 nm） were set at the maximum UV absorption wavelengths of these components. Twelve components （puerarin, daidzin, baicalin, wogonoside, liquiritin, berberine, palmatine, jatrorrhizine, glycyrrhizin, baicalein, wogonin and daidzein） were identified and determined using the developed method. All calibration curves showed good linear regression （r〉0.9995） within tested ranges. The injection precision, intra-day precisions and analysis repeatability were evaluated with the RSD values, which were no more than 0.97%, 1.69% and 1.71%, respectively. The recoveries were ranged from 97.4% to 100.2% with RSD values less than 1.87%. This readily available, low-cost and reliable HPLC-DAD method would improve the quality control of Gegen Qinlian pill.
本文首次建立了一种新的同时定性定量分析葛根芩连微丸中12种活性成分的高效液相色谱方法。采用Kromasil C_(18)色谱柱(250mm×4.6mm,5.0μm),以乙腈-0.02mol/L醋酸铵溶液(含0.3%三乙胺,1%冰醋酸调pH 4.3)为流动相,流速0.7mL/min,检测波长为250、280、346nm。本方法可同时定性和定量分析葛根素、大豆苷、大豆苷元、黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、小檗碱、巴马汀、药根碱、甘草苷、甘草酸等12种成分。各种成分在线性范围内线性均良好(r>0.9995);精密度、重复性、稳定性RSD值分别小于0.97%,1.69%和1.71%;回收率的范围为97.4%-100.2%,RSD值小于1.87%。所建方法简单、可行,可提高葛根芩连微丸的质量控制水平。
基金
National Natural Science Foundation of China (Grant No.30572376)
