Document Detail


Direct extract derivatization for determination of amino acids in human urine by gas chromatography and mass spectrometry.
MedLine Citation:
PMID:  12127324     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The purpose of this study was to develop a simple and accurate analytical method to determine amino acids in urine samples. The developed method involves the employment of an extract derivatization technique together with gas chromatography-mass spectrometry (GC-MS). Urine samples (300 microl) and an internal standard (10 microl) were placed in a screw tube. Ethylchloroformate (50 microl), methanol-pyridine (500 microl, 4:1, v/v) and chloroform (1 ml) were added to the tube. The organic layer (1 microl) was injected to a GC-MS system. In this proposed method, the amino acids in urine were derivatized during an extraction, and the analytes were then injected to GC-MS without an evaporation of the organic solvent extracted. Sample preparation was only required for ca. 5 min. The 15 amino acids (alanine, aspartic acid, cysteine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, tyrosine, tryptophan, valine) quantitatively determined in this proposed method. However, threonine, serine, asparagine, glutamine, arginine were not derivatized using any tested derivatizing reagent. The calibration curves showed linearity in the range of 1.0-300 microg/ml for each amino acid in urine. The correlation coefficients of the calibration curves of the tested amino acids were from 0.966 to 0.998. The limit of detection in urine was 0.5 microg/ml except for aspartic acid. This proposed method demonstrated substantial accuracy for detection of normal levels. This proposed method was limited for the determination of 15 amino acids in urine. However, the sample preparation was simple and rapid, and this method is suitable for a routine analysis of amino acids in urine.
Authors:
Akira Namera; Mikio Yashiki; Manami Nishida; Tohru Kojima
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences     Volume:  776     ISSN:  1570-0232     ISO Abbreviation:  J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.     Publication Date:  2002 Aug 
Date Detail:
Created Date:  2002-07-19     Completed Date:  2002-10-01     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101139554     Medline TA:  J Chromatogr B Analyt Technol Biomed Life Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  49-55     Citation Subset:  IM    
Copyright Information:
Copyright 2002 Elsevier Science B.V.
Affiliation:
Department of Legal Medicine, Hiroshima University Faculty of Medicine, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan. namera@hiroshima-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / urine*
Calibration
Gas Chromatography-Mass Spectrometry / methods*
Humans
Reference Standards
Reproducibility of Results
Chemical
Reg. No./Substance:
0/Amino Acids

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