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A novel UPLC–MS–MS method for simultaneous determination of seven uremic retention toxins with cardiovascular relevance in chronic kidney disease patients

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Abstract

Chronic kidney disease (CKD) is a devastating illness characterized by accumulation of uremic retention solutes in the body. The objective of this study was to develop and validate a simple, rapid, and robust UPLC–MS–MS method for simultaneous determination, in serum, of seven organic acid uremic retention toxins, namely uric acid (UA), hippuric acid (HA), indoxylsulfate (IS), p-cresylglucuronide (pCG), p-cresylsulfate (pCS), indole-3-acetic acid (IAA), and 3-carboxy-4-methyl-5-propyl-2-furanpropionic acid (CMPF). Isotopically labeled internal standards (d5-HA; 1,3-15N2-UA, and d5-IAA) were used to correct for variations in sample preparation and system performance. Separation on a C18 column was followed by negative electrospray ionization and tandem mass spectrometric detection. Accuracy was below the 15 % threshold. Within-day precision varied from 0.60 to 4.54 % and between-day precision was below 13.33 % for all compounds. The applicability of the method was evaluated by analyzing 78 serum samples originating both from healthy controls and from patients at different stages of CKD. These results were compared with those obtained by use of conventional HPLC–PDA–FLD methods. A good correlation was obtained between both methods for all compounds.

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Acknowledgments

Jente Boelaert gratefully acknowledges financial support from the Agency for Innovation in Science and Technology in Flanders (IWT).

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Authors and Affiliations

  1. Separation Science Group, Department of Organic Chemistry, Ghent University, Krijgslaan 281-S4bis, 9000, Ghent, Belgium

    Jente Boelaert, Frédéric Lynen & Pat Sandra

  2. Renal Division, Department of Internal Medicine, University Hospital Ghent, De Pintelaan 185, 9000, Ghent, Belgium

    Griet Glorieux, Sunny Eloot, Maria Van Landschoot, Marie-Anne Waterloos & Raymond Vanholder

  3. Research Institute for Chromatography, Kennedypark 26, 8500, Kortrijk, Belgium

    Pat Sandra

Authors
  1. Jente Boelaert
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  2. Frédéric Lynen
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  3. Griet Glorieux
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  4. Sunny Eloot
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  6. Marie-Anne Waterloos
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  7. Pat Sandra
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  8. Raymond Vanholder
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Correspondence to Frédéric Lynen.

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Boelaert, J., Lynen, F., Glorieux, G. et al. A novel UPLC–MS–MS method for simultaneous determination of seven uremic retention toxins with cardiovascular relevance in chronic kidney disease patients. Anal Bioanal Chem 405, 1937–1947 (2013). https://doi.org/10.1007/s00216-012-6636-9

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  • DOI: https://doi.org/10.1007/s00216-012-6636-9

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