Iothalamate quantification by tandem mass spectrometry to measure glomerular filtration rate

Jesse C. Seegmiller, Bradley E. Burns, Abdul H. Fauq, Naveen Mukhtar, John C Lieske, Timothy S. Larson

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

BACKGROUND: Glomerular filtration rate (GFR) can be determined by measuring renal clearance of the radio-contrast agent iothalamate. Current analytic methods for quantifying iothalamate concentrations in plasma and urine using liquid chromatography or capillary electrophoresis have limitations such as long analysis times and susceptibility to interferences. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to overcome these limitations. METHODS: Urine and plasma samples were deproteinized using acetonitrile and centrifugation. The supernatant was diluted in water and analyzed by LC-MS/MS using a water:methanol gradient. We monitored 4 multiple reaction monitoring transitions: m/z 614.8-487.0, 614.8-456.0, 614.8 -361.1, and 614.8 -177.1. We compared the results to those obtained via our standard capillary electrophoresis (CE-UV) on samples from 53 patients undergoing clinical GFR testing. RESULTS: Mean recovery was 90%-110% in both urine and plasma matrices. Imprecision was ≤15% for the m/z 614.8-487.0 and 614.8-456.0 transitions over a 10-day period at 1 mg/L. Method comparison for 159 patient samples (53 clearances) provided the following Passing-Bablok regressions: plasma iothalamate LC-MS/MS (y) vs CE-UV (x), y = 0.99x + 0.36; urine iothalamate LC-MS/MS vs CE-UV, y = 1.01x + 0.31; corrected GFR LC-MS/MS vs CE-UV, y = 1.00x + 0.00. Interfering substances prevented accurate iothalamate quantification by CE-UV in 2 patients, whereas these samples could be analyzed by LC-MS/MS. CONCLUSIONS: Iothalamate can be quantified by LCMS/MS for GFR measurement. This method circumvents potential problems with interfering substances that occasionally confound accurate GFR determinations.

Original languageEnglish (US)
Pages (from-to)568-574
Number of pages7
JournalClinical Chemistry
Volume56
Issue number4
DOIs
StatePublished - Apr 1 2010

Fingerprint

Iothalamic Acid
Tandem Mass Spectrometry
Glomerular Filtration Rate
Mass spectrometry
Urine
Plasmas
Capillary electrophoresis
Liquid chromatography
Capillary Electrophoresis
Liquid Chromatography
Centrifugation
Water
Radio
Contrast Media
Methanol
Kidney
Recovery
Monitoring
Testing

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Iothalamate quantification by tandem mass spectrometry to measure glomerular filtration rate. / Seegmiller, Jesse C.; Burns, Bradley E.; Fauq, Abdul H.; Mukhtar, Naveen; Lieske, John C; Larson, Timothy S.

In: Clinical Chemistry, Vol. 56, No. 4, 01.04.2010, p. 568-574.

Research output: Contribution to journalArticle

Seegmiller, Jesse C. ; Burns, Bradley E. ; Fauq, Abdul H. ; Mukhtar, Naveen ; Lieske, John C ; Larson, Timothy S. / Iothalamate quantification by tandem mass spectrometry to measure glomerular filtration rate. In: Clinical Chemistry. 2010 ; Vol. 56, No. 4. pp. 568-574.
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abstract = "BACKGROUND: Glomerular filtration rate (GFR) can be determined by measuring renal clearance of the radio-contrast agent iothalamate. Current analytic methods for quantifying iothalamate concentrations in plasma and urine using liquid chromatography or capillary electrophoresis have limitations such as long analysis times and susceptibility to interferences. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to overcome these limitations. METHODS: Urine and plasma samples were deproteinized using acetonitrile and centrifugation. The supernatant was diluted in water and analyzed by LC-MS/MS using a water:methanol gradient. We monitored 4 multiple reaction monitoring transitions: m/z 614.8-487.0, 614.8-456.0, 614.8 -361.1, and 614.8 -177.1. We compared the results to those obtained via our standard capillary electrophoresis (CE-UV) on samples from 53 patients undergoing clinical GFR testing. RESULTS: Mean recovery was 90{\%}-110{\%} in both urine and plasma matrices. Imprecision was ≤15{\%} for the m/z 614.8-487.0 and 614.8-456.0 transitions over a 10-day period at 1 mg/L. Method comparison for 159 patient samples (53 clearances) provided the following Passing-Bablok regressions: plasma iothalamate LC-MS/MS (y) vs CE-UV (x), y = 0.99x + 0.36; urine iothalamate LC-MS/MS vs CE-UV, y = 1.01x + 0.31; corrected GFR LC-MS/MS vs CE-UV, y = 1.00x + 0.00. Interfering substances prevented accurate iothalamate quantification by CE-UV in 2 patients, whereas these samples could be analyzed by LC-MS/MS. CONCLUSIONS: Iothalamate can be quantified by LCMS/MS for GFR measurement. This method circumvents potential problems with interfering substances that occasionally confound accurate GFR determinations.",
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AB - BACKGROUND: Glomerular filtration rate (GFR) can be determined by measuring renal clearance of the radio-contrast agent iothalamate. Current analytic methods for quantifying iothalamate concentrations in plasma and urine using liquid chromatography or capillary electrophoresis have limitations such as long analysis times and susceptibility to interferences. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to overcome these limitations. METHODS: Urine and plasma samples were deproteinized using acetonitrile and centrifugation. The supernatant was diluted in water and analyzed by LC-MS/MS using a water:methanol gradient. We monitored 4 multiple reaction monitoring transitions: m/z 614.8-487.0, 614.8-456.0, 614.8 -361.1, and 614.8 -177.1. We compared the results to those obtained via our standard capillary electrophoresis (CE-UV) on samples from 53 patients undergoing clinical GFR testing. RESULTS: Mean recovery was 90%-110% in both urine and plasma matrices. Imprecision was ≤15% for the m/z 614.8-487.0 and 614.8-456.0 transitions over a 10-day period at 1 mg/L. Method comparison for 159 patient samples (53 clearances) provided the following Passing-Bablok regressions: plasma iothalamate LC-MS/MS (y) vs CE-UV (x), y = 0.99x + 0.36; urine iothalamate LC-MS/MS vs CE-UV, y = 1.01x + 0.31; corrected GFR LC-MS/MS vs CE-UV, y = 1.00x + 0.00. Interfering substances prevented accurate iothalamate quantification by CE-UV in 2 patients, whereas these samples could be analyzed by LC-MS/MS. CONCLUSIONS: Iothalamate can be quantified by LCMS/MS for GFR measurement. This method circumvents potential problems with interfering substances that occasionally confound accurate GFR determinations.

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