Masár M, Kaniansky D, Bodor R, Jöhnck M, Stanislawski B
Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic.
J Chromatogr A. 2001 May 4;916(1-2):167-74. doi: 10.1016/s0021-9673(00)01094-3.
Isotachophoretic (ITP) separation and determination of a group of 13 organic and inorganic acids, currently present in wines, on a poly(methyl methacrylate) chip provided with on-column conductivity detection was a subject of a detailed study performed in this work. Experiments with the ITP electrolyte systems proposed to the separation of anionic constituents present in wine revealed that their separation at a low pH (2.9) provides the best results in terms of the resolution. Using a 94 mm long separation channel of the chip, the acids could be resolved within 10-15 min also in instances when their concentrations corresponded to those at which they typically occur in wines. A procedure suitable to the ITP determination of organic acids responsible for some important organoleptic characteristics of wines (tartaric, lactic, malic and citric acids) was developed. Concentrations of 2-10 mg/l of these acids represented their limits of quantitation for a 0.9 microl volume sample loop on the chip. A maximum sample load on the chip, under the preferred separating conditions, was set by the resolution of malate and citrate. A complete resolution of these constituents in wine samples was reached when their molar concentration ratio was 20:1 or less. ITP analyses of a large series of model and wine samples on the chip showed that qualitative indices [RSH (relative step height) values] of the acids, based on the response of the conductivity detector, reproduced with RSD better than 2% while reproducibilities of the determination of the acids of our interest characterized RSD values better than 3.5%.
采用配有柱上电导检测的聚甲基丙烯酸甲酯芯片,通过等速电泳(ITP)分离并测定葡萄酒中目前存在的一组13种有机和无机酸,是本工作中详细研究的主题。对用于分离葡萄酒中阴离子成分的ITP电解质系统进行的实验表明,在低pH值(2.9)下进行分离,在分离度方面能获得最佳结果。使用芯片94毫米长的分离通道,当酸的浓度与它们在葡萄酒中通常出现的浓度相对应时,这些酸也能在10 - 15分钟内得到分离。开发了一种适用于ITP测定对葡萄酒某些重要感官特性有贡献的有机酸(酒石酸、乳酸、苹果酸和柠檬酸)的方法。对于芯片上0.9微升体积的进样环,这些酸2 - 10毫克/升的浓度代表了它们的定量限。在优选的分离条件下,芯片上的最大进样量由苹果酸和柠檬酸的分离度决定。当葡萄酒样品中这些成分的摩尔浓度比为20:1或更低时,能实现它们的完全分离。在芯片上对大量模型样品和葡萄酒样品进行的ITP分析表明,基于电导检测器响应的酸的定性指标[相对步高(RSH)值]的重现性,相对标准偏差(RSD)优于2%,而我们感兴趣的酸的测定重现性的RSD值优于3.5%。