Determination of zeranol, taleranol, zearalanone, α-zearalenol, β-zearalenol and zearalenone in urine by LC-MS/MS

Abstract

An LC-MS/MS method was developed for the sensitive and selective determination of zeranol, taleranol, zearalanone, α-zearalenol, β-zearalenol and zearalenone in animal urine. Analysis was performed on the free compounds after enzymatic deconjugation. Sample preparation included liquid-liquid extraction followed by solid-phase extraction (SPE) with C18 and NH2 columns. For chromatographic separation of hormones an Inertsil(®) ODS-3 analytical column (150 mm × 2.1 mm, 3 µm) was used. The analytes were determined and identified by LC-MS/MS on a QTRAP5500 instrument with a TurboIon-Spray source operating in negative electrospray ionisation mode. For confirmatory purposes at least two transitions were obtained for each analyte. According to Commission Decision 2002 /657/EC the validation parameters - recovery, repeatability, reproducibility, linearity, specificity, decision limits and detection capabilities - were determined. All parameters were in agreement with 2002/657/EC performance criteria. The apparent recovery ranged from 76.2% to 116.3% for all examined compounds. The repeatability was below 20% and reproducibility did not exceed the limit of 25% for most analytes. Linearity was good for all analytes in the whole range of tested concentrations, as proved by the correlation coefficients greater than 0.99. The decision limits (CCα) ranged from 0.04 to 0.18 μg l(-1) for all analytes whereas the detection capabilities (CCβ) ranged from 0.07 to 0.31 μg l(-1), respectively. All CCα and CCβ values were below the recommended concentration of 2 μg l(-1). This analytical method will be used in an integrated system for Polish monitoring programmes for the confirmation of violative screened samples.

Determination of Trace Zearalenone and Its Metabolites in Human Serum by a High-Throughput UPLC-MS/MS Analysis

“…The mean concentration and range of each analyte were as following: ZEN (mean 0.31 ng/mL, range ND~3.68 ng/mL), β-ZOL (mean 0.084 ng/mL, range ND~1.32 ng/mL), α-ZOL (mean 0.051 ng/mL, range ND~2.64 ng/mL) and ZAN (mean 0.019 ng/mL, range ND~0.518 ng/mL). The results were comparable with those reported for other countries, normally with the mean concentration between 0.009~1.82 ng/mL [22,23,26,[32][33][34][35][36][37].…”

“…A number of approaches to quantify ZEN and its metabolites have been proposed for biological samples in last decade, which are commonly performed with gas or liquid chromatography coupled to mass or tandem mass spectrometry [19][20][21][22][23][24][25][26][27]. Among these methods, liquid chromatography-tandem mass spectrometry (LC-MS/MS) gradually became a prior technique for mycotoxin analysis in biological samples for its high selectivity and sensitivity.…”

Simultaneous determination of resorcylic acid lactones, β and α trenbolone and stilbenes in bovine urine by UHPLC/MS/MS

“…For example, Matraszek-Zuchowska et al [15] and Dusi et al [17] report the analysis of RALs in urine, Wozniak et al [16] the analysis of trenbolones, Kaklamanos et al [18] the analysis of stilbenes and RALs (excepting ␣-zearalenol and ␤-zearalenol) and Schmidt et al [19] the analysis of stilbenes and RALs. Regarding other matrices, several studies describe the determination of several compounds included in our work in meat [21,22], serum [23] and plasma [24,25].…”

“…These compounds can be analyzed directly (e.g. without derivatization) using High Performance Liquid Chromatography/Mass Spectrometry (HPLC/MS) and in recent years most publications in this field use this technique [15][16][17][18][19]. As with most HPLC/MS based methods a potential pitfall is the existence of matrix effects which can be minimized using the higher resolution provided by Ultra High Performance Liquid Chromatography/Mass Spectrometry (UHPLC/MS) [20].…”

Comparison of the Multiple Reaction Monitoring and Enhanced Product Ion Scan Modes for Confirmation of Stilbenes in Bovine Urine Samples Using LC–MS/MS QTRAP® System

“…A number of chromatographic methods, including gas chromatography (GC) and liquid chromatography (LC) in combination with different types of mass spectrometry (MS) have been demonstrated to determine hormones residues at low concentration levels in samples of animal origin [ 7 – 12 ]. In recent years, LC tandem mass spectrometry (MS/MS) with triple quadrupole (QqQ) mass analyzers operating in MRM scan mode was the dominant and powerful technique due to its high sensitivity and selectivity, used to quantify the targeted analytes such as drugs, steroids and pharmaceuticals at trace amounts in a variety of matrix [ 13 – 16 ]. From a technical point of view, in MRM mode, the two mass filtration steps are employed on a triple quadrupole mass spectrometer.…”