Scientific article
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Multitargeted Internal Calibration for the Quantification of Chronic Kidney Disease-Related Endogenous Metabolites Using Liquid Chromatography–Mass Spectrometry

Published inAnalytical chemistry, vol. 95, no. 36, p. 13546-13554
Publication date2023-09-01
First online date2023-09-01
Abstract

Accurate quantitative analysis in liquid chromatography–mass spectrometry (LC–MS) benefits from calibration curves generated in the same matrix as the study sample. In the case of endogenous compound quantification, as no blank matrix exists, the multitargeted internal calibration (MTIC) is an attractive and straightforward approach to avoid the need for extensive matrix similarity evaluation. Its principle is to take advantage of stable isotope labeled (SIL) standards as internal calibrants to simultaneously quantify authentic analytes using a within sample calibration. An MTIC workflow was developed for the simultaneous quantification of metabolites related to chronic kidney disease (CKD) using a volumetric microsampling device to collect 20 μL of serum or plasma, followed by a single-step extraction with acetonitrile/water and liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. Since a single concentration of internal calibrant is necessary to calculate the study sample concentration, the instrument response function was investigated to determine the best SIL concentration. After validation, the trueness of 16 endogenous analytes in authentic human serum ranged from 72.2 to 116.0%, the repeatability from 1.9 to 11.3%, and the intermediate precision ranged overall from 2.1 to 15.4%. The proposed approach was applied to plasma samples collected from healthy control participants and two patient groups diagnosed with CKD. Results confirmed substantial concentration differences between groups for several analytes, including indoxyl sulfate and cortisone, as well as metabolite enrichment in the kynurenine and indole pathways. Multitargeted methodologies represent a major step toward rapid and straightforward LC–MS/MS absolute quantification of endogenous biomarkers, which could change the paradigm of MS use in clinical laboratories.

Keywords
  • Calibration
  • Chromatography, Liquid
  • Humans
  • Renal Insufficiency, Chronic / diagnosis
  • Tandem Mass Spectrometry
Citation (ISO format)
VISCONTI, Gioele et al. Multitargeted Internal Calibration for the Quantification of Chronic Kidney Disease-Related Endogenous Metabolites Using Liquid Chromatography–Mass Spectrometry. In: Analytical chemistry, 2023, vol. 95, n° 36, p. 13546–13554. doi: 10.1021/acs.analchem.3c02069
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ISSN of the journal0003-2700
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Creation03/10/2023 20:14:13
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