Liquid chromatography/electrospray ionization tandem mass spectrometric screening and confirmation methods for beta2-agonists in human or equine urine.
Abstract: Electrospray ionization (ESI) mass spectra of 19 common beta(2)-agonists were investigated in terms of fragmentation pattern and dissociation behavior of the analytes, proving the origin of fragment ions and indicating mechanisms of charge-driven and charge-remote fragmentation. Based on these data, liquid chromatographic/ESI tandem mass spectrometric (LC/ESI-MS/MS) screening and confirmation methods were developed for doping control purposes. These procedures employ established sample preparation steps including either acidic or enzymatic hydrolysis, alkaline extraction and, in the case of equine urine specimens, acidic re-extraction of the analytes. In addition, a degradation product of formoterol caused by acidic hydrolysis during sample preparation could be identified and utilized as target compound in screening and also confirmation methods. The screening procedures cover 18 or 19beta(2)-agonists, the estimated limits of detection of which for equine and human urine samples vary between 2 and 100 ng ml(-1) and between 2 and 50 ng ml(-1), respectively. A single LC/MS/MS analysis can be performed in 9 min.
Copyright 2003 John Wiley & Sons, Ltd.
Publication Date: 2003-12-04 PubMed ID: 14648827DOI: 10.1002/jms.542Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The researchers conducted an investigation on the electrospray ionization (ESI) mass spectra of 19 common beta2-agonists, focusing on their fragmentation and behavioral dissociation. The data obtained was used to formulate liquid chromatographic/ESI tandem mass spectrometric (LC/ESI-MS/MS) methodologies that could test for the presence of these beta2-agonists in human and equine urine for doping control purposes.
Fragmentation and Dissociation Behavior
- The experiment was undertaken to understand in detail how beta2-agonists break down and separate under the influence of ESI.
- Particular emphasis was laid on understanding the charge-driven and charge-remote fragmentation mechanisms of the analytes.
- This deep understanding of fragmentation and dissociation of beta2-agonists under ESI was used in developing a reliable detection method based on liquid chromatography and ESI tandem mass spectrometry (LC/ESI-MS/MS).
Development of LC/ESI-MS/MS Screening Method
- Using information obtained from the fragmentation and dissociation behavior of beta2-agonists, the LC/ESI-MS/MS screening and confirmation methods were developed.
- This screening method uses acidic or enzymatic hydrolysis, alkaline extraction and, in the case of equine urine samples, an acidic re-extraction of the analytes, as part of its sample preparation steps.
- An interesting finding was the identification of a degradation product of formoterol that occurs due to acidic hydrolysis during sample preparation. This compound could be utilized as a target in the screening and confirmation methodology.
Performance of the Developed Screening Method
- The method was effective in detecting between 18 or 19 beta2-agonists.
- The estimated limits of detection for equine and human urine samples were found to vary between 2 and 100 ng ml(-1) and between 2 and 50 ng ml(-1), respectively.
- In terms of duration, a single LC/ESI-MS/MS analysis using this methodology can be completed within 9 minutes, indicating the potential for efficient, high-throughput screening.
Cite This Article
APA
Thevis M, Opfermann G, Schänzer W.
(2003).
Liquid chromatography/electrospray ionization tandem mass spectrometric screening and confirmation methods for beta2-agonists in human or equine urine.
J Mass Spectrom, 38(11), 1197-1206.
https://doi.org/10.1002/jms.542 Publication
Researcher Affiliations
- Institute of Biochemistry, German Sport University, Cologne, Carl-Diem Weg 6, 50933 Cologne, Germany. m.thevis@biochem.dshs-koeln.de
MeSH Terms
- Adrenergic beta-Agonists / chemistry
- Adrenergic beta-Agonists / metabolism
- Adrenergic beta-Agonists / urine
- Animals
- Artifacts
- Chromatography, Liquid / methods
- Clenbuterol / analogs & derivatives
- Clenbuterol / chemistry
- Clenbuterol / metabolism
- Clenbuterol / urine
- Doping in Sports
- Ethanolamines / chemistry
- Ethanolamines / metabolism
- Ethanolamines / urine
- Formoterol Fumarate
- Horses / urine
- Humans
- Mass Screening / methods
- Molecular Structure
- Sensitivity and Specificity
- Spectrometry, Mass, Electrospray Ionization / methods
Citations
This article has been cited 3 times.- Hammouda MEA, Salem YA, El-Ashry SM, El-Enin MAA. Isocratic ion pair chromatography for estimation of novel combined inhalation therapy that blocks coronavirus replication in chronic asthmatic patients.. Sci Rep 2023 Jan 6;13(1):305.
- Marlier D. Doping in Racing Pigeons (Columba livia domestica): A Review and Actual Situation in Belgium, a Leading Country in This Field.. Vet Sci 2022 Jan 22;9(2).
- Kim HS, Siluk D, Wainer IW. Quantitative determination of fenoterol and fenoterol derivatives in rat plasma using on-line immunoextraction and liquid chromatography/mass spectrometry.. J Chromatogr A 2009 Apr 17;1216(16):3526-32.
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