Determination of amino acids in rat vitreous perfusates by capillary electrophoresis
Determination of amino acids in rat vitreous perfusates by capillary electrophoresis
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DOI：
10.1002/elps.200405941
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发表时间：
2004-09-01
期刊：
影响因子：
2.9
通讯作者：
Shippy, SA
中科院分区：
文献类型：
--
作者：
Thongkhao-On, K;Kottegoda, S;Shippy, SA
氨基酸的体内测定对于提高我们对生物组织功能和功能障碍的生理状态的理解是重要的。然而，不同生物流体的化学复杂基质使这类重要分子的测定复杂化。我们介绍了一种方法，用于表征亚微升体积的玻璃体液灌注液的氨基酸组成。低流量推挽式灌注采样与在复杂基质中收集小体积样本兼容，这些样本可能难以分离。采用3-（4-羧基苯甲酰基）-2-喹啉-甲醛（CBQCA）衍生化和毛细管电泳（CE）分离-激光诱导荧光检测（LIF）技术，建立了一种高效、灵敏、快速分析玻璃体灌注液中氨基酸的方法。衍生与CBQCA长达2小时提供了高灵敏度和低检测限在nm级。包括D-丝氨酸（D-Ser）和D-天冬氨酸（D-Asp）在内的17种氨基酸在不到10分钟的时间内被解析。用这种测定技术表征玻璃体氨基酸将有助于理解眼部疾病和视觉生理机制。
In vivo determinations of amino acids are important for improving our understanding of physiological states of biological tissue function and dysfunction. However, the chemically complex matrix of different biological fluids complicates the assay of this important class of molecules. We introduce a method for characterizing the amino acid composition of submicroliter volumes of vitreous humor perfusates. Low-flow push-pull perfusion sampling is compatible with collecting small volume samples in a complicated matrix that are potentially difficult to separate. An efficient, sensitive, and rapid analysis of amino acids from in vivo perfusates of the vitreous is presented with 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) derivatitation and capillary electrophoresis (CE) separation with laser-induced fluorescence detection (LIF). Derivatization with CBQCA for up to 2 h provided high sensitivity and low detection limits at the nm level. Seventeen amino acids including D-serine (D-Ser) and D-aspartate (D-Asp) were resolved in less than 10 min. Importantly, D-Ser is separated from its enantiomeric pair. Characterization of vitreal amino acids with this assay technique will be useful for understanding ocular diseases and physiological mechanisms in vision.