On-capillary derivatization and analysis of amino acids in human plasma by capillary electrophoresis with laser-induced fluorescence detection: application to diagnosis of aminoacidopathies.

Full Abstract

A capillary electrophoresis with laser-induced fluorescence detection method for the analysis of free amino acids (AA) in human plasma was developed. A mixture of 16 AA was on-capillary derivatized with 3-(2-furoyl)quinoline-2-carboxaldehyde (FQ) and separated inside the capillary in less than 30 min using 70 mM borax-3.5 mM SDS pH 9.3 as running buffer. Four plasma samples from a healthy donor and patients suffering from phenylketonuria, propionic acidemia, and tyrosinemia type II were studied. Repeatabilities calculated as intra-day RSD (n = 3) values for the AA involved in these aminoacidopathies (glycine, phenylalanine, and tyrosine) were in the range of 0.3 to 1.2% for migration time and 3.7 to 8.2% for peak height. Reproducibilities calculated as inter-day RSD (n = 4) values for the same AA were between 0.7 and 1.4% for migration time and 4.7 and 9.1% for peak height. A fast qualitative analysis allowed the identification of the corresponding disease by comparing the electrophoretic profiles from the patient and the healthy donor and noting the increased level of the specific AA accompanying each individual disease. The results of the quantitative analysis for glycine, phenylalanine, and tyrosine in the plasma samples studied using the developed method showed a good agreement with those provided by the Center of Diagnosis of Molecular Diseases using a standard method for AA analysis.

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Author information

Author/s: Veledo, Maria Teresa (MT); de Frutos, Mercedes (M); Diez-Masa, Jose Carlos (JC);

Affiliation: Institute of Organic Chemistry (C.S.I.C.), Juan de la Cierva 3, E-28006 Madrid, Spain.

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't

Journal: Electrophoresis (Electrophoresis), published in Germany. (Language: eng)

Reference: 2006-Aug; vol 27 (issue 15) : pp 3101-7

Dates: Created 2006/08/28; Completed 2006/12/12;

PMID: 16800027, status: MEDLINE (last retrieval date: 12/26/2008)

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Amino Acid Metabolism, Inborn Errors - blood; diagnosis
Amino Acids - blood; isolation & purification
Case-Control Studies
Electrophoresis, Capillary - methods

Lasers
Reference Standards
Reproducibility of Results
Spectrometry, Fluorescence - methods

Associated Chemicals: Amino Acids (0)

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