Chromatographic estimation of iothalamate and p-aminohippuric acid to measure glomerular filtration rate and effective renal plasma flow in humans

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Abstract

Iothalamate (IOT) clearance and p-aminohippuric acid (PAH) clearance are used for estimation of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). A simple and rapid method is described for simultaneous determination of IOT and PAH in the same chromatogram in the serum and urine of humans. The mobile phase consisted of methanol-50 mM sodium monobasic phosphate with 0.5 mM tetrabutyl ammonium hydrogen sulfate (18:82, v/v), pumped at a rate of 0.8 ml/min on a C18 reversed-phase column. Samples of serum and urine were deproteinized with two volumes of acetonitrile containing the internal standard, p-aminobenzoic acid (PABA). The UV detector was set at 254 nm and peak height ratios of PAH or IOT to PABA were calculated with an integrator. Precision and accuracy were within 15% for both PAH and IOT. The recovery of PAH in urine and serum were 94% and 91%, respectively. For IOT the corresponding recoveries were 93% and 92%, respectively. This method clearly distinguishes acetyl-PAH from PAH and has been validated in healthy volunteers.

Original languageEnglish (US)
Pages (from-to)3-9
Number of pages7
JournalJournal of Chromatography B: Biomedical Applications
Volume705
Issue number1
StatePublished - Jan 23 1998
Externally publishedYes

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Iothalamic Acid
p-Aminohippuric Acid
Plasma flow
4-Aminobenzoic Acid
Ultraviolet detectors
Recovery
Ammonium Compounds
Sulfates
Methanol
Hydrogen

Keywords

  • Iothalamate
  • p-Aminohippuric acid

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

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title = "Chromatographic estimation of iothalamate and p-aminohippuric acid to measure glomerular filtration rate and effective renal plasma flow in humans",
abstract = "Iothalamate (IOT) clearance and p-aminohippuric acid (PAH) clearance are used for estimation of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). A simple and rapid method is described for simultaneous determination of IOT and PAH in the same chromatogram in the serum and urine of humans. The mobile phase consisted of methanol-50 mM sodium monobasic phosphate with 0.5 mM tetrabutyl ammonium hydrogen sulfate (18:82, v/v), pumped at a rate of 0.8 ml/min on a C18 reversed-phase column. Samples of serum and urine were deproteinized with two volumes of acetonitrile containing the internal standard, p-aminobenzoic acid (PABA). The UV detector was set at 254 nm and peak height ratios of PAH or IOT to PABA were calculated with an integrator. Precision and accuracy were within 15{\%} for both PAH and IOT. The recovery of PAH in urine and serum were 94{\%} and 91{\%}, respectively. For IOT the corresponding recoveries were 93{\%} and 92{\%}, respectively. This method clearly distinguishes acetyl-PAH from PAH and has been validated in healthy volunteers.",
keywords = "Iothalamate, p-Aminohippuric acid",
author = "Rajiv Agarwal",
year = "1998",
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language = "English (US)",
volume = "705",
pages = "3--9",
journal = "Journal of Chromatography B: Biomedical Sciences and Applications",
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T1 - Chromatographic estimation of iothalamate and p-aminohippuric acid to measure glomerular filtration rate and effective renal plasma flow in humans

AU - Agarwal, Rajiv

PY - 1998/1/23

Y1 - 1998/1/23

N2 - Iothalamate (IOT) clearance and p-aminohippuric acid (PAH) clearance are used for estimation of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). A simple and rapid method is described for simultaneous determination of IOT and PAH in the same chromatogram in the serum and urine of humans. The mobile phase consisted of methanol-50 mM sodium monobasic phosphate with 0.5 mM tetrabutyl ammonium hydrogen sulfate (18:82, v/v), pumped at a rate of 0.8 ml/min on a C18 reversed-phase column. Samples of serum and urine were deproteinized with two volumes of acetonitrile containing the internal standard, p-aminobenzoic acid (PABA). The UV detector was set at 254 nm and peak height ratios of PAH or IOT to PABA were calculated with an integrator. Precision and accuracy were within 15% for both PAH and IOT. The recovery of PAH in urine and serum were 94% and 91%, respectively. For IOT the corresponding recoveries were 93% and 92%, respectively. This method clearly distinguishes acetyl-PAH from PAH and has been validated in healthy volunteers.

AB - Iothalamate (IOT) clearance and p-aminohippuric acid (PAH) clearance are used for estimation of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). A simple and rapid method is described for simultaneous determination of IOT and PAH in the same chromatogram in the serum and urine of humans. The mobile phase consisted of methanol-50 mM sodium monobasic phosphate with 0.5 mM tetrabutyl ammonium hydrogen sulfate (18:82, v/v), pumped at a rate of 0.8 ml/min on a C18 reversed-phase column. Samples of serum and urine were deproteinized with two volumes of acetonitrile containing the internal standard, p-aminobenzoic acid (PABA). The UV detector was set at 254 nm and peak height ratios of PAH or IOT to PABA were calculated with an integrator. Precision and accuracy were within 15% for both PAH and IOT. The recovery of PAH in urine and serum were 94% and 91%, respectively. For IOT the corresponding recoveries were 93% and 92%, respectively. This method clearly distinguishes acetyl-PAH from PAH and has been validated in healthy volunteers.

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