title: A liquid chromatography - tandem mass spectrometry method to measure a selected panel of uremic retention solutes derived from endogenous and colonic microbial metabolism.
abstract: Chronic kidney disease (CKD) is associated with an increased risk of mortality and cardiovascular disease, which is, at least partly, mediated by the accumulation of so-called uremic retention solutes. Although there has been an increasing interest in the behavior of these solutes, derived from both the endogenous and colonic microbial metabolism, methods to simultaneously and accurately measure a broad panel of relevant uremic retention solutes remain scarce. We developed a highly sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. A high throughput sample preparation was used with extraction of analytes from 50 μl serum using Ostro plate technology. For most solutes, stable isotopes labelled metabolites were used as internal standards. Chromatography was achieved using an Acquity UPLC CSH Fluoro Phenyl column. The total run time was 8 min, the mobile phase was a gradient of 0.1% formic acid in Milli-Q water and pure methanol at a flow rate of 0.5 ml min(-1). Detection was performed using a tandem mass spectrometer with alternated positive and negative electrospray ionization. Calibration curves were linear for all solutes. Precision was assessed according to the NCCLS EP5-T guideline, being below 15% for all metabolites. Mean recoveries were between 83 and 104% for all metabolites. The validated method was successfully applied in a cohort of 488 patients with CKD. We developed and validated a sensitive and robust UPLC-MS/MS method for quantification of 15 uremic retention solutes derived from endogenous and colonic microbial metabolism. This method allows for studying the behavior and relevance of these solutes in patients with CKD.
authors: KU Leuven - University of Leuven, Department of Microbiology and Immunology, Laboratory of Nephrology, B-3000 Leuven, Belgium.
authors: Henriette
authors: H
authors: de Loor
authors: KU Leuven - University of Leuven, Department of Microbiology and Immunology, Laboratory of Nephrology, B-3000 Leuven, Belgium.
authors: Ruben
authors: R
authors: Poesen
authors: KU Leuven - University of Leuven, Department of Chemistry, Division of Molecular Design and Synthesis, B-3000 Leuven, Belgium.
authors: Wout
authors: W
authors: De Leger
authors: KU Leuven - University of Leuven, Department of Chemistry, Division of Molecular Design and Synthesis, B-3000 Leuven, Belgium.
authors: Wim
authors: W
authors: Dehaen
authors: KU Leuven - University of Leuven, Department of Pharmaceutical and Pharmacological Sciences, Drug Delivery and Disposition, B-3000 Leuven, Belgium.
authors: Patrick
authors: P
authors: Augustijns
authors: KU Leuven - University of Leuven, Department of Microbiology and Immunology, Laboratory of Nephrology, B-3000 Leuven, Belgium; University Hospitals Leuven, Department of Nephrology and Renal Transplantation, B-3000 Leuven, Belgium.
authors: Pieter
authors: P
authors: Evenepoel
authors: KU Leuven - University of Leuven, Department of Microbiology and Immunology, Laboratory of Nephrology, B-3000 Leuven, Belgium; University Hospitals Leuven, Department of Nephrology and Renal Transplantation, B-3000 Leuven, Belgium. Electronic address: bjorn.meijers@uzleuven.be.
authors: Björn
authors: B
authors: Meijers
mesh_terms: D001774:Blood Chemical Analysis; D002851:Chromatography, High Pressure Liquid; D003106:Colon / Q000378:metabolism* / Q000382:microbiology; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood / Q000378:metabolism* / Q000382:microbiology; D006435:Renal Dialysis; D053719:Tandem Mass Spectrometry
keywords: Indoxyl sulfate; Phenylacetylglutamine; Tandem mass spectrometry; Trimethylamine-N-oxide; Ultra-performance liquid chromatography; Uremic retention solutes; p-Cresyl sulfate