Simultaneous determination of ibuprofen and its metabolites in complex equine urine matrices by GC-EI-MS in excretion study in view of doping control.
Abstract: A novel assay for the simultaneous determination of ibuprofen (IBU) and its four probable metabolites, 1-hydroxyibuprofen (1-OH IBU), 2-hydroxyibuprofen (2-OH IBU), 3-hydroxyibuprofen (3-OH IBU) and carboxyibuprofen (CBX IBU) in equine urine samples with the application of Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) has been developed and elaborated. The new approach for sample preparation including minimizing matrix effects by the application of weak cation exchange solid-phase extraction together with strong cation exchange solid-phase extraction has been applied. The GC-EI-MS method was validated to demonstrate specificity, matrix effect, linearity, limit of detection (LOD) and quantification (LOQ), precision, trueness, carry-over and stability by using the matrix-matched quality control samples. Additionally, extraction yield was evaluated. The assay achieved the LOQ of 1.75 μg mL, 0.62 μg mL, 4.15 μg mL, 0.58 μg mL and 4.04 μg mL for IBU, 1-OH IBU, 2-OH IBU, 3-OH IBU and CBX IBU, respectively. The developed method has been successfully applied to the excretion study in horses, in which a single oral IBU dose was administered to twelve horses (mares and geldings) and equine urine samples were collected for 5 or 6 days after the drug administration. Data on the detection and determination of three IBU metabolites, 2-OH IBU, 3-OH IBU and CBX IBU in equine urine samples has been presented for the first time. The obtained results indicated the rapid excretion of IBU and its metabolites that were detectable only in the first day after the drug administration. IBU was mainly the most abundant compound detected in equine urine samples (with two exceptions in the case of samples collected from two horses, for which the highest instrumental responses were obtained for CBX IBU). The received results have indicated that two major IBU metabolites, CBX IBU and 2-OH IBU can be important markers for the IBU abuse in view of doping control in equestrian sports.
Copyright © 2018 Elsevier B.V. All rights reserved.
Publication Date: 2018-02-05 PubMed ID: 29453024DOI: 10.1016/j.jpba.2018.02.004Google Scholar: Lookup
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Summary
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The research focused on creating a new testing method for detecting ibuprofen and its metabolites in horse urine, which can be an important tool for preventing doping in equestrian sports.
Development of the Assay
- The novel assay was developed for the simultaneous detection of ibuprofen (IBU) and its four potential metabolites, including 1-hydroxyibuprofen (1-OH IBU), 2-hydroxyibuprofen (2-OH IBU), 3-hydroxyibuprofen (3-OH IBU), and carboxyibuprofen (CBX IBU) in equine urine samples.
- Highly sensitive Gas Chromatography-Electron Ionization-Mass Spectrometry (GC-EI-MS) was used to measure the presence and quantity of these substances.
- The team implemented a new approach for sample preparation to minimize matrix effects, which includes weak cation exchange solid-phase extraction together with strong cation exchange solid-phase extraction.
Assay Validation
- The GC-EI-MS method underwent rigorous validation to prove its specificity, matrix effect, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, trueness, carry-over, and stability.
- Validation was done using matrix-matched quality control samples, which are known quantities of the analytes embedded in an identical matrix to the sample.
- The extraction yield, or the amount of IBU and its metabolites extracted from the sample, was also evaluated.
Application of Assay in Excretion Study
- The method was then employed in a study that observed the excretion of ibuprofen and its metabolites by twelve horses over five to six days following a single dose of the drug.
- All metabolites were found to be rapidly excreted, and were only detectable the day after the drug administration, indicating a short window of detection.
- Ibuprofen was, with only two exceptions, the most abundant compound found in the urine samples.
- The study also showed that the metabolites CBX IBU and 2-OH IBU could be important markers of ibuprofen usage, and therefore useful in the detection of doping in equestrian sports.
Cite This Article
APA
Waraksa E, Wójtowicz-Zawadka M, Kwiatkowska D, Jarek A, Małkowska A, Wrzesień R, Namieśnik J.
(2018).
Simultaneous determination of ibuprofen and its metabolites in complex equine urine matrices by GC-EI-MS in excretion study in view of doping control.
J Pharm Biomed Anal, 152, 279-288.
https://doi.org/10.1016/j.jpba.2018.02.004 Publication
Researcher Affiliations
- Gdańsk University of Technology, Faculty of Chemistry, Department of Analytical Chemistry, G. Narutowicz 11/12 Street, 80-233 Gdansk, Poland; Institute of Sport - National Research Institute, Trylogii 2/16 Street, 01-982 Warsaw, Poland.
- Institute of Sport - National Research Institute, Department of Anti-Doping Research, Trylogii 2/16 Street, 01-982 Warsaw, Poland. Electronic address: marzena.wojtowicz@insp.waw.pl.
- Institute of Sport - National Research Institute, Department of Anti-Doping Research, Trylogii 2/16 Street, 01-982 Warsaw, Poland.
- Institute of Sport - National Research Institute, Department of Anti-Doping Research, Trylogii 2/16 Street, 01-982 Warsaw, Poland.
- Medical University of Warsaw, Department of Toxicology, Banacha 1 Street, 02-097 Warsaw, Poland.
- Medical University of Warsaw, Central Laboratory of Experimental Animal, Banacha 1B Street, 02-097 Warsaw, Poland.
- Gdańsk University of Technology, Faculty of Chemistry, Department of Analytical Chemistry, G. Narutowicz 11/12 Street, 80-233 Gdansk, Poland.
MeSH Terms
- Animals
- Doping in Sports / methods
- Gas Chromatography-Mass Spectrometry / methods
- Horses
- Ibuprofen / analogs & derivatives
- Ibuprofen / metabolism
- Ibuprofen / urine
- Limit of Detection
- Solid Phase Extraction / methods
Citations
This article has been cited 2 times.- Shiri S, Alizadeh K, Abbasi N. A novel technique for simultaneous determination of drugs using magnetic nanoparticles based dispersive micro-solid-phase extraction in biological fluids and wastewaters. MethodsX 2020;7:100952.
- Piechocka J, Matwiej N, Gaweł M, Matyjaszczyk M, Głowacki R, Chwatko G. Application of the HPLC-ELSD technique for the determination of major metabolites of ibuprofen and creatinine in human urine. Sci Rep 2023 Nov 20;13(1):20268.
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