title: Determination of uremic solutes in biological fluids of chronic kidney disease patients by HPLC assay.
abstract: During chronic kidney disease (CKD), solutes called uremic solutes, accumulate in blood and tissues of patients. We developed an HPLC method for the simultaneous determination of several uremic solutes of clinical interest in biological fluids: phenol (Pol), indole-3-acetic acid (3-IAA), p-cresol (p-C), indoxyl sulfate (3-INDS) and p-cresol sulfate (p-CS). These solutes were separated by ion-pairing HPLC using an isocratic flow and quantified with a fluorescence detection. The mean serum concentrations of 3-IAA, 3-INDS and p-CS were 2.12, 1.03 and 13.03 μM respectively in healthy subjects, 3.21, 17.45 and 73.47 μM in non hemodialyzed stage 3-5 CKD patients and 5.9, 81.04 and 120.54 μM in hemodialyzed patients (stage 5D). We found no Pol and no p-C in any population. The limits of quantification for 3-IAA, 3-INDS, and p-CS were 0.83, 0.72, and 3.2 μM respectively. The within-day CVs were between 1.23 and 3.12% for 3-IAA, 0.98 and 2% for 3-INDS, and 1.25 and 3.01% for p-CS. The between-day CVs were between 1.78 and 5.48% for 3-IAA, 1.45 and 4.54% for 3-INDS, and 1.19 and 6.36% for p-CS. This HPLC method permits the simultaneous and quick quantification of several uremic solutes for daily analysis of large numbers of samples.
authors: Laboratoire de Biochimie Moléculaire Fondamentale et Clinique, UFR de Pharmacie, Université de la Méditerranée, 27 boulevard Jean Moulin, 13 385 Marseille cedex 5, France. raymond.calaf@univmed.fr
authors: Raymond
authors: R
authors: Calaf
authors: Claire
authors: C
authors: Cerini
authors: Cécile
authors: C
authors: Génovésio
authors: Pierre
authors: P
authors: Verhaeghe
authors: Noémie
authors: N
authors: Jourde-Chiche
authors: David
authors: D
authors: Bergé-Lefranc
authors: Bertrand
authors: B
authors: Gondouin
authors: Laetitia
authors: L
authors: Dou
authors: Sophie
authors: S
authors: Morange
authors: Angel
authors: A
authors: Argilés
authors: Pascal
authors: P
authors: Rathelot
authors: Françoise
authors: F
authors: Dignat-George
authors: Philippe
authors: P
authors: Brunet
authors: Philippe
authors: P
authors: Charpiot
mesh_terms: D000368:Aged; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003408:Cresols / Q000097:blood*; D005260:Female; D006801:Humans; D007200:Indican / Q000097:blood*; D007210:Indoleacetic Acids / Q000097:blood*; D007676:Kidney Failure, Chronic / Q000097:blood*; D008297:Male; D008875:Middle Aged; D019800:Phenol / Q000097:blood*; D010636:Phenols / Q000097:blood*; D013463:Sulfuric Acid Esters / Q000097:blood*; D014511:Uremia / Q000097:blood*
chemical_list: D003408:Cresols; D007210:Indoleacetic Acids; D010636:Phenols; D013463:Sulfuric Acid Esters; D019800:Phenol; C408690:4-cresol sulfate; C030737:indoleacetic acid; D007200:Indican