Depletion of urinary zilpaterol residues in horses as measured by ELISA and UPLC-MS/MS.
Abstract: Three horses were dosed with dietary zilpaterol and the urine concentrations measured from withdrawal day 0 to withdrawal day 21. The analyses were carried out using both enzyme-linked immunosorbent assay (ELISA) and an ultraperformance liquid chromatography with triple-quadrupole-tandem mass spectrometric detection (UPLC-MS/MS). The UPLC-MS/MS method was developed to provide rapid analysis with positive analyte identification by following three product ions and computing the two independent ion ratios. When urinary zilpaterol concentrations were between 0.2 and 2 ng/mL, the ELISA had interday recoveries of 114-120% with coefficients of variation (CV) of <22%; intraday recoveries were 79-111% with CVs of <13%. For urinary zilpaterol concentrations of 0.4-40 ng/mL the UPLC-MS/MS method had interday recoveries of 94-104% with CVs of <8%; intraday recoveries were 97-102% with CVs of < or = 7.5%. Correlation analysis demonstrated that the ELISA and UPLC-MS/MS methods returned essentially the same results, especially at urinary zilpaterol concentrations below 2000 ng/mL. Urinary excretion peaked rapidly after dosing between 5300 and 10800 ng/mL (UPLC-MS/MS) or between 5900 and 17900 ng/mL (ELISA) for the different horses, much higher than observed in other species. Urinary zilpaterol concentrations declined rapidly to below 3000 ng/mL within 24 h of study day 1. After about 5 days, zilpaterol elimination slowed markedly, taking nearly 10 days for an order of magnitude decrease. The analytical methods were able to detect zilpaterol in the urine even at withdrawal day 21, demonstrating the sensitivity of each analytical method and the slow rate of zilpaterol depuration from horses.
Publication Date: 2010-03-12 PubMed ID: 20218607DOI: 10.1021/jf904253tGoogle Scholar: Lookup
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Summary
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This research tracked the residue of dietary zilpaterol in horse urine over a period of 21 days using two types of measurement methods: ELISA and UPLC-MS/MS. It found that the residues decline slowly over time, indicating the slow rate of zilpaterol expulsion from the body of horses.
Methods Used
- The study was conducted on three horses that were fed dietary zilpaterol. Urine samples were collected and tested from withdrawal day 0 until day 21.
- The two methods used for urine analysis were ELISA (Enzyme-Linked Immunosorbent Assay) and UPLC-MS/MS (Ultra Performance Liquid Chromatography with Triple Quadrupole Tandem Mass Spectrometric Detection). Both the methods were developed for rapid analysis and positive identification of the analyte.
Findings of the Study
- The study found that both the ELISA and UPLC-MS/MS methods returned similar results, especially when the urinary zilpaterol concentrations were below 2000 ng/mL.
- The urinary excretion of zilpaterol reached its peak quickly after dosing. The peak values varied between 5300 and 10800 ng/mL (UPLC-MS/MS) or 5900 and 17900 ng/mL (ELISA) depending on the horse. These values were much higher than those observed in other species.
- Within 24 hours from the start of the study, urinary zilpaterol concentrations rapidly decreased to below 3000 ng/mL. After about 5 days, the elimination of zilpaterol slowed down considerably, taking approximately 10 days for a tenfold decrease.
- Both the analytical methods used in the study were able to detect zilpaterol in the horse urine even on withdrawal day 21, demonstrating the high sensitivity of both methods and the slow rate of zilpaterol depuration from horses.
Significance of the Study
- This study provides valuable information on the elimination rate of zilpaterol in horses and the sensitivity of both ELISA and UPLC-MS/MS methods in tracking zilpaterol concentrations in urine. The findings can be useful for doping tests and drug regulation in horse racing.
Cite This Article
APA
Shelver WL, Thorson JF, Hammer CJ, Smith DJ.
(2010).
Depletion of urinary zilpaterol residues in horses as measured by ELISA and UPLC-MS/MS.
J Agric Food Chem, 58(7), 4077-4083.
https://doi.org/10.1021/jf904253t Publication
Researcher Affiliations
- USDA-ARS, Biosciences Research Laboratory, Fargo, North Dakota 58105, USA. weilin.shelver@ars.usda.gov
MeSH Terms
- Animals
- Drug Residues / analysis
- Enzyme-Linked Immunosorbent Assay / methods
- Horses
- Tandem Mass Spectrometry / methods
- Trimethylsilyl Compounds / urine
- Veterinary Drugs / urine
Citations
This article has been cited 2 times.- Machin J, Brewer K, Morales-Briceno A, Fenger C, Maylin G, Tobin T. Sporadic worldwide "clusters" of feed driven Zilpaterol identifications in racing horses: a review and analysis.. Ir Vet J 2022 May 14;75(1):11.
- Cheng TD, Shelver WL, Hong CC, McCann SE, Davis W, Zhang Y, Ambrosone CB, Smith DJ. Urinary Excretion of the β-Adrenergic Feed Additives Ractopamine and Zilpaterol in Breast and Lung Cancer Patients.. J Agric Food Chem 2016 Oct 12;64(40):7632-7639.
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