目的:建立一测多评法同时测定蒙药述达格-4 总黄酮提取物中槲皮素、山柰酚、高良姜素、山柰素、高良姜素-3-甲醚5 个黄酮类成分的含量。方法:采用Agilent C18 色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇(B)-0.5% 乙酸水溶液(A),梯度洗脱,流速0.6 mL·min-1,柱温30 ℃,进样量20 μL,检测波长： 286 nm 检测山柰酚、山柰素、高良姜素-3-甲醚,256 nm 检测槲皮素。以高良姜素为内参,计算其他4 个黄酮类成分的相对校正因子,采用该相对校正因子计算其他4 个成分的含量,实现一测多评。同时采用外标法测定5 个黄酮类成分的含量,并对二者测定结果进行比较,验证一测多评法的可行性和准确性。结果:在一定线性范围内,以高良姜素为内参,槲皮素、山柰酚、山柰素、高良姜素-3-甲醚4 个黄酮类成分的平均校正因子分别为0.094 8、0.226 6、0.267 0、0.155 4,且一测多评法测定结果与外标法测定结果无显著差异。结论:建立了同时测定蒙药述达格-4 总黄酮提取物中5 个黄酮类成分含量的一测多评法,该法快速、准确、简便、可靠,可用于蒙药述达格-4 的质量控制研究。
Objective: To establish a quantitative analysis of multi-components by single marker(QAMS)method for simultaneous determination of five flavonoids(quercetin,kaempferol,galangin,kaempferide,and galangin-3-methylether)in total flavonoids extract of Mongolian medicine Shudage-4. Methods: HPLC analysis was performed on an Agilent C18 column(4.6 mm×250 mm,5 μm)with gradient elution of methanol(B)-0.5% acetic acid in water(A)at a flow rate of 0.6 mL·min-1. The column temperature was 30 ℃ and the injection volume was 20 μL. The detection wavelength was set at 254 nm for quercetin and kaempferol,286 nm for kaempferol,galangin,kaempferide and galangin-3-methylether. Galangin was selected as the internal reference substance,the relative correction factors (RCFs)between galangin and the other fourflavonoids were established,and the contents of the five constituents in samples were calculated to realize QAMS.At the same time,the external standard method was used to quantify contents of the five constituents,and the two methods were compared to validate the accuracy and feasibility of the new method. Results: Within a certain linear range,the RCFs of quercetin,kaempferol,kaempferide and galangin-3-methylether were 0.094 8,0.226 6,0.267 0,and 0.155 4 respectively. With galangin as the internal reference,there was no significant difference between the quantitative results of QAMS and external standard method(ESM). Conclusion: The AQAMS method for the simultaneous determination of five flavones in total flavonoids extract of Mongolian medicine Shudage-4 was established. This method is rapid,accurate,simple and reliable,and can be used for the quality control study of Mongolian medicine Shudage-4.
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