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Analysis of free and bound O-phosphoamino acids in urine by gas chromatography with flame photometric detection.
- Source :
-
Biomedical chromatography : BMC [Biomed Chromatogr] 1993 Jul-Aug; Vol. 7 (4), pp. 184-8. - Publication Year :
- 1993
-
Abstract
- A selective and sensitive method has been developed for the analysis of free and bound forms of O-phosphoamino acids, such as O-phosphoserine, O-phosphothreonine and O-phosphotyrosine, in urine samples by gas chromatography (GC). For free O-phosphoamino acid analysis, the urine sample was extracted with trichloroacetic acid and run through an ion exchange column. For total (free plus bound) O-phosphoamino acid analysis, the urine sample was hydrolysed in acid and base in order to release the O-phosphoamino acid from peptides or proteins. O-Phosphoamino acids in these prepared samples were converted into their N-isobutoxycarbonyl trimethyl ester derivatives and then measured by GC with flame photometric detection (FPD-GC). The calibration curve was linear in the range of 10-500 ng for each O-phosphoamino acid, and the detection limits were 30-80 pg as injection amounts. O-Phosphoamino acids in the urine samples could be selectively determined by the FPD-GC method without any influence from coexisting substances. The recoveries of O-phosphoamino acids added to urines and urine hydrolysates were 90-98% and relative standard deviations were 1.5-8.0%. By using this method, we investigated an age dependence of O-phosphoamino acid excretion in normal subjects.
- Subjects :
- Animals
Chromatography, Gas
Chromatography, Ion Exchange
Diet
Gas Chromatography-Mass Spectrometry
Humans
Hydrolysis
Milk
Phosphoserine urine
Phosphothreonine urine
Phosphotyrosine
Photometry
Protein Binding
Trichloroacetic Acid
Tyrosine analogs & derivatives
Tyrosine urine
Amino Acids urine
Organophosphorus Compounds urine
Subjects
Details
- Language :
- English
- ISSN :
- 0269-3879
- Volume :
- 7
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Biomedical chromatography : BMC
- Publication Type :
- Academic Journal
- Accession number :
- 7693088
- Full Text :
- https://doi.org/10.1002/bmc.1130070403