High-performance liquid chromatography of the renal blood flow marker p-aminohippuric acid (PAH) and its metabolite N-acetyl PAH improves PAH clearance measurements.

Details

Serval ID
serval:BIB_FDA84FF15C25
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
High-performance liquid chromatography of the renal blood flow marker p-aminohippuric acid (PAH) and its metabolite N-acetyl PAH improves PAH clearance measurements.
Journal
Journal of chromatography. B, Biomedical sciences and applications
Author(s)
Decosterd L.A., Karagiannis A., Roulet J.M., Bélaz N., Appenzeller M., Buclin T., Vogel P., Biollaz J.
ISSN
1387-2273
Publication state
Published
Issued date
1997
Volume
703
Number
1-2
Pages
25-36
Language
english
Notes
Publication types: Comparative Study ; Journal Article - Publication Status: ppublish
Abstract
PAH (N-(4-aminobenzoyl)glycin) clearance measurements have been used for 50 years in clinical research for the determination of renal plasma flow. The quantitation of PAH in plasma or urine is generally performed by colorimetric method after diazotation reaction but the measurements must be corrected for the unspecific residual response observed in blank plasma. We have developed a HPLC method to specifically determine PAH and its metabolite NAc-PAH using a gradient elution ion-pair reversed-phase chromatography with UV detection at 273 and 265 nm, respectively. The separations were performed at room temperature on a ChromCart (125 mmx4 mm I.D.) Nucleosil 100-5 microm C18AB cartridge column, using a gradient elution of MeOH-buffer pH 3.9 1:99-->15:85 over 15 min. The pH 3.9 buffered aqueous solution consisted in a mixture of 375 ml sodium citrate-citric acid solution (21.01 g citric acid and 8.0 g NaOH per liter), added up with 2.7 ml H3PO4 85%, 1.0 g of sodium heptanesulfonate and completed ad 1000 ml with ultrapure water. The N-acetyltransferase activity does not seem to notably affect PAH clearances, although NAc-PAH represents 10.2+/-2.7% of PAH excreted unchanged in 12 healthy subjects. The performance of the HPLC and the colorimetric method have been compared using urine and plasma samples collected from healthy volunteers. Good correlations (r=0.94 and 0.97, for plasma and urine, respectively) are found between the results obtained with both techniques. However, the colorimetric method gives higher concentrations of PAH in urine and lower concentrations in plasma than those determined by HPLC. Hence, both renal (ClR) and systemic (Cls) clearances are systematically higher (35.1 and 17.8%, respectively) with the colorimetric method. The fraction of PAH excreted by the kidney ClR/ClS calculated from HPLC data (n=143) is, as expected, always <1 (mean=0.73+/-0.11), whereas the colorimetric method gives a mean extraction ratio of 0.87+/-0.13 implying some unphysiological values (>1). In conclusion, HPLC not only enables the simultaneous quantitation of PAH and NAc-PAH, but may also provide more accurate and precise PAH clearance measurements.
Keywords
Biological Markers, Calibration, Chromatography, High Pressure Liquid, Circadian Rhythm, Colorimetry, Humans, Linear Models, Male, Renal Circulation, Reproducibility of Results, Sensitivity and Specificity, Spectrophotometry, Ultraviolet, p-Aminohippuric Acid
Pubmed
Web of science
Create date
25/01/2008 10:41
Last modification date
20/08/2019 16:28
Usage data