毛细管前衍生和毛细管区带电泳分析饮料中的氨基酸

Apichai Santalad, Pattana Teerapornchaisit, Rodjana Burakham, Supalax Srijaranai
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引用次数: 7

摘要

建立了一种简便、快速的氨基酸同时分析方法。以1,2-萘醌-4-磺酸盐(NQS)为原料,在碱性介质(pH 10.0)中进行毛细管前衍生化,并在70℃下进行反应。用毛细管区带电泳(CZE)分析了它们的衍生物。考察了缓冲液(pH、类型和浓度)、有机改性剂和分离电压等参数对CZE分离的影响。最佳条件为硼酸盐(pH 10.0) 70 mmol L-1,含10%乙腈,分离电压12 kV,进样(0.5 psi, 5s), 240 nm毛细管上检测。7种氨基酸的分离在17 min内完成,所有氨基酸的检出限为1.0 mg L-1。在1.0 ~ 100.0 mg L-1的浓度范围内,标定曲线呈线性关系。迁移时间重复性≤1.0%,日间重复性≤2.0%,峰面积重复性≤5.0%,日间重复性≤6.0%。该方法已应用于几种饮料样品,在衍生化前仅对样品进行了简单的稀释和过滤处理,并通过CZE进行了分析。
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Pre-Capillary Derivatisation and Capillary Zone Electrophoresis for Amino Acids Analysis in Beverages

A simple and rapid method for the simultaneous analysis of amino acids has been developed. Amino acids were derivatised based on pre-capillary derivatisation with 1,2-naphthoquinone-4-sulfonate (NQS) in basic medium (pH 10.0) and developed reaction at 70 °C. Their derivatives were analysed by capillary zone electrophoresis (CZE). The parameters affecting CZE separation were investigated including buffer (pH, type and concentration), organic modifier and separation voltage. The optimum condition was 70 mmol L-1 borate (pH 10.0) containing 10 % acetonitrile, separation voltage of 12 kV, and sample injection (0.5 psi, 5s) and on-capillary detection at 240 nm. The separation of seven amino acids was achieved within 17 min. The detection limit was 1.0 mg L-1 for all studied amino acids. The calibration curves were linear in the concentration range of 1.0 – 100.0 mg L-1. The repeatability, intra-day and inter-day analysis were ≤ 1.0 % and ≤ 2.0 % for migration time and ≤ 5.0 % and 6.0 % for peak area. The proposed method has been applied to several beverage samples with only a simple dilution and filtration treatment of sample before derivatisation and analysed by CZE.

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