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Characterization of a reversed-phase high-performance liquid chromatographic system for the determination of blood amino acids
- Source :
- Journal of chromatography. 507
- Publication Year :
- 1990
-
Abstract
- High-performance liquid chromatography was used to separate physiological amino acids in perchloric acid supernatants of blood samples. Precolumn derivatization with phenyl isothiocyanate was carried out, starting with 20 microliters of supernatant; 2-10 microliters were injected into a 30-cm Pico Tag column, which was eluted with a gradient of two eluents in 64 min. Stock amino acid solutions prepared in water, hydrochloric acid or perchloric acid showed comparable recoveries on serial dilution (parallelism test). The recovery of crystalline amino acids added to blood in amounts ranging from normal to six times normal was generally satisfactory. The within-assay relative standard deviations were less than 5% for many amino acids. The performance of the system was less than satisfactory for cysteine and methionine. Glutamine and asparagine are interconverted into glutamate and aspartate, respectively, in a time-dependent fashion; a separate measurement of one member of the pair is therefore required in order to assay the other starting from the sum of both chromatographic peaks. The method is suitable for the relatively rapid, sensitive and accurate measurement of blood amino acids in perchloric acid supernatants (in which other relevant metabolites are customarily assayed) over a wide range of physiological concentrations, on very small amounts of sample.
- Subjects :
- chemistry.chemical_classification
Chromatography
Perchlorates
Serial dilution
Phenyl isothiocyanate
Elution
Organic Chemistry
Hydrochloric acid
General Medicine
Blood Proteins
Biochemistry
Analytical Chemistry
Amino acid
chemistry.chemical_compound
chemistry
Isothiocyanates
Humans
Perchloric acid
Amino Acids
Chromatography, High Pressure Liquid
Thiocyanates
Precolumn derivatization
Subjects
Details
- Volume :
- 507
- Database :
- OpenAIRE
- Journal :
- Journal of chromatography
- Accession number :
- edsair.doi.dedup.....4b8ca35d6991256ebb12fdd773cc01f2