Determination of amino acids in rat vitreous perfusates by capillary electrophoresis

Kongthong Thongkhao-On, Sumith Kottegoda, Jose S. Pulido, Scott A. Shippy

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2978-2984
Number of pages7
JournalELECTROPHORESIS
Volume25
Issue number17
DOIs
StatePublished - Sep 1 2004

Keywords

  • Amino acids
  • Chiral separation
  • Flourescence
  • Small-volume samples
  • Vitreous samples

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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