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目的优化红车轴草提取物的定量测定方法,以客观有效地评价提取物的质量。方法以鹰嘴豆芽素A、染料木素、刺芒柄花素和大豆苷元4种异黄酮苷元作为定量测定的指标成分,采用HPLC法分别测定提取物酸水解前后异黄酮的量。结果通过系统的研究与优化,确定的色谱条件为:Capcell pak C18柱(150 mm×4.6 mm,5μm),柱温30℃,检测波长254 nm,体积流量1.0 mL/min,以乙腈(含0.05%TFA)-水(含0.05%TFA)梯度洗脱;酸水解条件为:提取物以1 mol/L盐酸甲醇溶液(每毫克提取物样品加2.0 mL)于85℃水浴回流1.5 h,冷却至室温后定容待测。结论本方法为红车轴草提取物提供了一种简便快速、有效可靠的定量测定方法。
Objective To optimize the method of quantitative determination of extract of Trifolium repens to evaluate the quality of extract objectively and effectively. Methods Four kinds of isoflavone aglycones, such as chickpea sp. A, genistein, thorn formonon and daidzein, were used as index components. The content of isoflavones before and after acid hydrolysis was determined by HPLC. Results The chromatographic conditions were as follows: capcell pak C18 column (150 mm × 4.6 mm, 5 μm), column temperature 30 ℃, detection wavelength 254 nm, volume flow rate 1.0 mL / min, % TFA) -water (containing 0.05% TFA). The acid hydrolysis conditions were as follows: the extract was refluxed in 85 ℃ water bath for 1.5 h in 1 mol / L methanolic hydrochloric acid solution After room temperature to be measured. Conclusion This method provides a simple, rapid, effective and reliable method for the quantitative determination of red clover extract.