A screening and determinative method for the analysis of natural and synthetic steroids, stilbenes and resorcyclic acid lactones in bovine urine

Abstract

Our laboratory has four separate methods for the analysis of trenbolone, stilbenes, resorcyclic acid lactones, and estradiol in bovine urine. The method described in this paper was in response to a client request to consolidate the methods preferably into one method. A multiresidue semi-quantitative method was developed and any suspect positive samples detected by the method were subjected to the method of standard addition to accurately quantify the concentration of the positive analyte. Samples were enzymatically hydrolyzed using β-glucuronidase after which, supported liquid extraction on HM-N cartridges was performed, followed by solvent exchange into methyl tert-butyl ether (MTBE). The samples were evaporated and reconstituted into 10% methanol in water and loaded onto a SampliQ OPT SPE. The cleaned-up extract was further cleaned up on a SampliQ NH 2 cartridge. The SPE eluate was split into two for analysis by gas chromatography-mass spectrometry (GC-MS) using electron ionization (EI) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) using both positive and negative electrospray. It was found that with the exception of estradiol and trenbolone all compounds could be analyzed by both GC-MS and LC-MS/MS, providing a semi-quantitative method. It is recommended that quantification is achieved using standard addition. Of the 13 compounds successfully monitored, the limits of detection (LODs), and the limits of quantification (LOQs) obtained were within the Codex limits for the target concentrations. As far as the authors are aware, the use of supported liquid extraction has not been reported for bovine urine analysis.

A Quantitative and Confirmatory Method Employing Liquid Chromatography Coupled to Hybrid High-Resolution Mass Spectrometry and QuEChERS for the Determination of Thirty-Seven Growth Promoter Residues in Bovine Urine

“…Liquid chromatography (LC) coupled with mass spectrometry (MS) is frequently employed due to the vast working concentration ranges and to the wide variety of compounds to be investigated. − Gas chromatography (GC) coupled with MS has also been employed; ,, however, an additional derivatization step for non-volatile compounds is frequently required. Low-resolution mass spectrometry (LRMS) has accounted for most of the works regarding multiresidue methods due to the higher sensitivity and instrumental robustness. − ,− The use of high-resolution mass spectrometry (HRMS), especially the hybrid spectrometers with a combination between LRMS and HRMS, has been increasingly studied, e.g. Q-Orbitrap, a quadrupole-orbitrap hybrid mass spectrometer. ,, …”

“…As for sample preparation, solid phase extraction (SPE) ,− , and a combination between SPE and liquid–liquid extraction (LLE) ,,,,, were among the most frequently used procedures. Other works used successive LLE, LLE along with a derivatization step, and one work applied the QuEChERS approach…”

Preface to the proceedings of the SASKVAL III international workshop on validation and regulatory analysis

“…In the second paper submitted under this theme, Akre and Mizuno demonstrated how difficult it is to develop a single method for the analysis and detection of natural and synthetic steroids, stilbenes, and resorcylic acid lactones in bovine urine despite recent advances made in the detection capabilities of current platform technologies such as gas chromatography‐tandem mass spectrometry (GC‐MS/MS) and liquid chromatography‐tandem mass spectrometry (LC‐MS/MS).…”

Proceedings SaskVal 2015

“…Following publication of Proceedings SaskVal 2015 in Drug Testing and Analysis Volume 8 Issues 5‐6 May‐June 2016, the following corrections and updates have been applied:

Tourism Saskatoon/Gordon Taylor has been given credit for the photo used as the Cover image of the issue.The Accreditation and Disclaimer statements: The Workshop was sponsored by the OECD Co‐operative Research Programme: Biological Resource Management for Sustainable Agricultural Systems, whose financial support made it possible for some of the invited speakers to participate in the Workshop. Disclaimer: The opinions expressed and arguments employed in this publication are the sole responsibility of the authors and do not necessarily reflect those of the OECD or of the governments of its Member countries. have been inserted in the Table of Contents and Editorial

Funding statement: Travel by the Lead Author to present this manuscript at SASKVAL III was funded by the OECD Co‐operative Research Programme: Biological Resource Management for Sustainable Agricultural Programmes. has been updated/added and Disclaimer has been included in papers by Jack F. Kay, Christine Akre, Bjorn J.A. Berendsen, Anton Kaufmann, John D.G.

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“…has been updated/added and Disclaimer has been included in papers by Jack F. Kay, Christine Akre, Bjorn J.A. Berendsen, Anton Kaufmann, John D.G.…”