DETERMINATION OF UREA KINETICS BY ISOTOPE-DILUTION WITH [C-13]UREA AND GAS-CHROMATOGRAPHY ISOTOPE RATIO MASS-SPECTROMETRY (GC-IRMS) ANALYSIS

Citation
Wd. Kloppenburg et al., DETERMINATION OF UREA KINETICS BY ISOTOPE-DILUTION WITH [C-13]UREA AND GAS-CHROMATOGRAPHY ISOTOPE RATIO MASS-SPECTROMETRY (GC-IRMS) ANALYSIS, Clinical science, 93(1), 1997, pp. 73-80
Citations number
16
Categorie Soggetti
Medicine, Research & Experimental
Journal title
ISSN journal
01435221
Volume
93
Issue
1
Year of publication
1997
Pages
73 - 80
Database
ISI
SICI code
0143-5221(1997)93:1<73:DOUKBI>2.0.ZU;2-X
Abstract
1. Stable urea isotopes can be used to study urea kinetics in humans, The use of stable urea isotopes far studying urea kinetic parameters i n humans on a large scale is hampered by the high costs of the labelle d material, We devised a urea dilution for measurement of the distribu tion volume, production rate and clearance of urea in healthy subjects and renal failure patients using the inexpensive single labelled [C-1 3]urea isotope with subsequent analysis by headspace chromatography-is otope ratio MS (GC-IRMS) of the [C-13]urea enrichment, 2. The method i nvolves measurement of the molar percentage excess of [C-13]urea in pl asma samples taken over a 4 h period after an intravenous bolus inject ion of [C-13]urea, During the sample processing procedure, the plasma samples together with calibration samples containing a known molar per centage excess of [C-13]urea are acidified with phosphoric acid to rem ove endogenous CO2, and are subsequently incubated with urease to conv ert the urea present in the plasma samples into CO2. The C-13 enrichme nt of the generated CO2 is analysed by means of GC-IRMS, This method a llows measurement of the molar percentage excess of [C-13]urea to an a ccuracy of 0.02%. 3. Reproducibility studies showed that the sample pr ocessing procedure [within-run coefficient of variation (CV) <2.8% and between-run CV <8.8%] and the GC-IRMS analysis (within-day CV <1.3% a nd between-day CV <1.3%) could be repeated with good reproducibility, 4. In clinical urea kinetic studies in a healthy subject and in a rena l failure patient without residual renal function, reproducible values of the distribution volume, production rate and clearance of urea wer e determined using minimal amounts of [C-13]urea (25-50 mg). 5. becaus e only low [C-13]urea enrichments are needed in this urea dilution met hod using GC-IRMS analysis, the costs of urea kinetic studies are redu ced considerably, especially in patients with renal failure.