title: Validation of a UHPLC-MS/MS method for quantification of zearalenone, α-zearalenol, β-zearalenol, α-zearalanol, β-zearalanol and zearalanone in human urine.
abstract: Humans can be exposed to mycotoxins through the diet. Evaluation of exposure levels to mycotoxins can be performed by direct determination in urine. The present work proposes a sensitive ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of zearalenone (ZON) and its five metabolites (α-zearalenol [α-ZOL], β-zearalenol [β-ZOL], α-zearalanol [zeranol, α-ZAL], β-zearalanol [teranol, β-ZAL] and zearalanone [ZAN]) in human urine samples. The method involves the enzymatic hydrolysis of the samples, extraction of the analytes using liquid-liquid extraction (LLE) with ethyl acetate/formic acid (99:1 v/v) and a cleanup step using hexane, prior to their quantification by UHPLC-MS/MS, using an electrospray ionization (ESI) interface in the negative mode. Zearalenone-d6 (ZON-d6) was used as surrogate. The limits of detection and the limits of quantification ranged from 0.03 to 0.3ngmL(-1) and from 0.1 to 1.0ngmL(-1), respectively. The method was validated using matrix-matched calibration and a spike recovery assay. Recovery rates for spiked samples ranged from 96% to 104%, with relative standard deviations lower than 8.5%. This method was satisfactorily applied to 42 urine samples from Tunisian women for the determination of zearalenone and its five metabolites.
authors.affiliation: Center of Urgent Medical Assistance of Tunis, Laboratory of Toxicology and Environment (LR12SP07), Tunis, Tunisia.
authors.forename: H
authors.initials: H
authors.lastname: Belhassen
authors.affiliation: Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, E-18012, Spain; Laboratory of Medical Investigations, San Cecilio University Hospital, University of Granada, Granada, E-18071, Spain. Electronic address: ijdiaz@ugr.es.
authors.forename: I
authors.initials: I
authors.lastname: Jiménez-Díaz
authors.affiliation: Center of Urgent Medical Assistance of Tunis, Laboratory of Toxicology and Environment (LR12SP07), Tunis, Tunisia.
authors.forename: R
authors.initials: R
authors.lastname: Ghali
authors.affiliation: Center of Urgent Medical Assistance of Tunis, Laboratory of Toxicology and Environment (LR12SP07), Tunis, Tunisia.
authors.forename: H
authors.initials: H
authors.lastname: Ghorbel
authors.affiliation: Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, E-18012, Spain; Laboratory of Medical Investigations, San Cecilio University Hospital, University of Granada, Granada, E-18071, Spain; CIBER en Epidemiología y Salud Pública (CIBERESP), Granada, E-18071, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Molina-Molina
authors.affiliation: Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, E-18012, Spain; Laboratory of Medical Investigations, San Cecilio University Hospital, University of Granada, Granada, E-18071, Spain; CIBER en Epidemiología y Salud Pública (CIBERESP), Granada, E-18071, Spain.
authors.forename: N
authors.initials: N
authors.lastname: Olea
authors.affiliation: Center of Urgent Medical Assistance of Tunis, Laboratory of Toxicology and Environment (LR12SP07), Tunis, Tunisia.
authors.forename: A
authors.initials: A
authors.lastname: Hedili
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D009183:Mycotoxins / Q000652:urine*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D015025:Zearalenone / Q000652:urine*; D015029:Zeranol / Q000031:analogs & derivatives* / Q000652:urine
chemical_list: D009183:Mycotoxins; C029659:zearalenol; D015025:Zearalenone; D015029:Zeranol
keywords: Human urine samples; Mycotoxins; Tunisian women; UHPLC–MS/MS; Zearalenone; Zearalenone metabolites