Document Detail

Automated simultaneous isolation and quantitation of labeled amino acid fractions from plasma and tissue by ion-exchange chromatography.
MedLine Citation:
PMID:  7866514     Owner:  NLM     Status:  MEDLINE    
In order to trace metabolic pathways of amino acids in the body, a known labeled amount of an amino acid is infused. Dilution in the body pool is measured, using the specific activity and calculated by dividing the labeled amount of an amino acid (tracer) by its total pool (tracer + tracee). This paper describes a method, which combines fractionation and quantitation of multiple amino acids in one chromatographic run. To achieve this, we performed a classical amino acid ion-exchange separation on standard HPLC equipment. The column effluent was divided continuously into two solvent streams using a rapidly switching, pump controlled "split-valve". The main part (90%) was directed to a computer controlled fraction collector, while the remaining 10% was mixed with o-phthaldialdehyde reagent after which fluorescence was measured. Using this system, 10-1000 microliters of deproteinized plasma, representing a maximum of 50 nmol of each amino acid, could be fractionated and quantitated in the same chromatographic run. In addition to optimal counting efficiency of an off-line radioactivity counter, it enabled easy measurement of the specific activity of multiple amino acid tracers.
H M van Eijk; M P Huinck; D R Rooyakkers; N E Deutz
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of chromatography. B, Biomedical applications     Volume:  660     ISSN:  1572-6495     ISO Abbreviation:  J. Chromatogr. B, Biomed. Appl.     Publication Date:  1994 Oct 
Date Detail:
Created Date:  1995-03-29     Completed Date:  1995-03-29     Revised Date:  2007-10-16    
Medline Journal Info:
Nlm Unique ID:  9421796     Medline TA:  J Chromatogr B Biomed Appl     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  251-7     Citation Subset:  IM    
Department of Surgery, University of Limburg, Maastricht, Netherlands.
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MeSH Terms
Amino Acids / blood*
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Indicators and Reagents
Spectrometry, Fluorescence
Reg. No./Substance:
0/Amino Acids; 0/Indicators and Reagents; 643-79-8/o-Phthalaldehyde

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