Fully automated assay for total homocysteine, cysteine, cysteinylglycine, glutathione, cysteamine, and 2-mercaptopropionylglycine in plasma and urine

We describe a 6-min HPLC method to measure the total concentrations of the most important thiols in plasma and urine--cysteine, homocysteine, cysteinylglycine, and glutathione--as well as the concentrations in plasma and urine, respectively, of cysteamine and 2-mercaptopropionylglycine, two compound...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 1998-04, Vol.44 (4), p.825-832
Main Authors: Pastore, Anna, Massoud, Renato, Motti, Corradino, Russo, Anna Lo, Fucci, Giorgio, Cortese, Claudio, Federici, Giorgio
Format: Article
Language:English
Subjects:
Adolescent
Adult
Amino Acids, Sulfur - blood
Amino Acids, Sulfur - urine
Analytical biochemistry: general aspects, technics, instrumentation
Analytical, structural and metabolic biochemistry
Autoanalysis - methods
Biological and medical sciences
Child
Child, Preschool
Chromatography, High Pressure Liquid - methods
Cysteamine - blood
Cysteamine - urine
Cysteine - blood
Cysteine - urine
Dipeptides - blood
Dipeptides - urine
Female
Fundamental and applied biological sciences. Psychology
Glutathione - blood
Glutathione - urine
Homocysteine - blood
Homocysteine - urine
Humans
Infant
Infant, Newborn
Investigative techniques, diagnostic techniques (general aspects)
Male
Medical sciences
Miscellaneous. Technology
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Reference Values
Sensitivity and Specificity
Sulfhydryl Compounds - blood
Sulfhydryl Compounds - urine
Tiopronin - blood
Tiopronin - urine
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Summary:We describe a 6-min HPLC method to measure the total concentrations of the most important thiols in plasma and urine--cysteine, homocysteine, cysteinylglycine, and glutathione--as well as the concentrations in plasma and urine, respectively, of cysteamine and 2-mercaptopropionylglycine, two compounds used to treat disorders of cysteine metabolism. Precolumn derivatization with bromobimane and reversed-phase HPLC were performed automatically by a sample processor. Throughput was up to 100 samples in 24 h. The within-run CV ranged from 0.9% to 3.4% and the between-run CV ranged from 1.5% to 6.1%. Analytical recovery was 97-107%, with little difference between plasma and urine samples. The detection limit was approximately 50 nmol/L for all the analytes studied. Thiol concentrations were determined in the plasma of 206 healthy donors and in the urine of 318 healthy donors distributed for age and sex. Mean values of plasma cysteine and homocysteine were significantly lower in infants (ages,
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/44.4.825