高效液相色谱法测定木瓜水溶性有机酸含量的样品处理方法研究

Sample processing method for determination of water soluble organic acids in Chaenomelis Fructus by HPLC

目的优化木瓜水溶性有机酸含量测定的样品处理方法。方法采用经验证的高效液相色谱法,以苹果酸、莽草酸、绿原酸、原儿茶酸、奎尼酸、咖啡酸含量为评判指标,用单因素试验法考察提取方式(回流和超声提取),乙醇浓度和料液比对有机酸提取量的影响;提取后经净化、富集;用流动相中的水相溶解、进样分析。结果样品最佳处理方法为50%乙醇、料液比为1∶20、超声提取60 min;在此条件下,木瓜有机酸提取量较高,非酸性成分干扰较少。色谱图基线平直,6种水溶性有机酸的色谱峰对称性和分离度良好。结论该方法能有效提取样品中水溶性有机酸,降低HPLC分析中的基质效应,可为木瓜有机酸含量测定提供稳定可靠的供试品溶液制备方法。

Objective To optimize the sample processing method for determination of water soluble organic acids in Chaenomelis Fructus by a validated HPLC -UV method.Methods Quinic acid,malic acid,shikimic acid,protocatechuic acid,chlorogenic acid and caffeic acid were determined and used as an evaluation parameter.Single factor tests method was applied for evaluating the extrac-tion（reflux and ultrasonic -assisted extraction）,ethanol concentration and the effect of ratio of material to liquid on extraction of or-ganic acids.The extracting solution with further purification and enrichment in mobile phase （A）employed were analyzed.Results The optimum method was as follows：ratio of material to liquid 1：20 and ethanol concentration 50% with ultrasonic -assisted extraction for 60 min.Under optimal conditions,the high extraction efficiency of the organic acids in Chaenomelis Fructus was obtained with less interference from non -acidic components of the sample.A straight baseline,chromatographic peak symmetry and good resolution for the six kinds of water soluble organic acids were conducted in the chromatogram.Conclusions This sample processing method is ef-fective,stable and reliable for extraction of water soluble organic acids in Chaenomelis Fructus,which can reduce the matrix effect sig-nificantly in the HPLC analysis.