Comprehensive screening of anabolic steroids, corticosteroids, and acidic drugs in horse urine by solid-phase extraction and liquid chromatography-mass spectrometry.
Abstract: This paper reports two highly efficient liquid chromatography-mass spectrometry (LC-MS) methods for the screening of anabolic steroids, corticosteroids, and acidic drugs for the purpose of doping control in equine sports. Sample extraction was performed using a mixed-mode C8-SCX solid-phase extraction (SPE) cartridge. The first eluted fraction (acidic/neutral fraction) was base-washed and the resulting organic extract was used for the screening of anabolic steroids and corticosteroids by LC-MS using multiple reaction monitoring (MRM) in the positive electrospray ionisation (ESI) mode. The remaining aqueous extract was re-adjusted to pH 6 and acidic drugs were recovered by liquid/liquid extraction. Detection was again achieved using LC-MRM but in the negative ESI mode. A total of 40 anabolic steroids and corticosteroids, and over 50 acidic drugs, including some cyclooxygenase-2 (COX-2) inhibitors, oxicams, anti-diabetics, sedatives, diuretics and Delta(9)-tetrahydro-11-norcannabinol-9-carboxylic acid, could be covered by the two LC-MS methods. Both methods utilized a high efficiency reversed-phase column (3.3 cm L x 2.1 mm I.D. with 3 microm particles) coupled with a fast-scanning triple-quadrupole mass spectrometer to achieve fast turnaround times. The overall turnaround times for both methods were 10 min, inclusive of post-run and equilibration times.
Publication Date: 2006-05-02 PubMed ID: 16631183DOI: 10.1016/j.chroma.2006.03.089Google Scholar: Lookup
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- Journal Article
Summary
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The study is on two innovative liquid chromatography-mass spectrometry methods for screening doping substances such as anabolic steroids, corticosteroids and acidic drugs in horse urine. These methods ensure efficient and fast detection with a total turnaround time of 10 minutes.
Research Methods
- The researchers have used two effective liquid chromatography-mass spectrometry (LC-MS) techniques for testing doping substances in horseracing.
- For sample extraction, they utilized a mixed-mode C8-SCX solid-phase extraction (SPE) cartridge. This tool aids in segregating and concentrating particular components from the samples.
- The study relied on multiple reaction monitoring (MRM) in positive electrospray ionisation (ESI) mode for the initial fraction screening of anabolic steroids and corticosteroids.
- The continuing aqueous extract was fine-tuned to pH 6 and acidic drugs were recuperated using liquid/liquid extraction. They used LC-MRM again but in negative ESI mode for detection.
Detection Span
- A broad spectrum of over 90 substances was detected using these methods. This includes corticosteroids and anabolic steroids, acidic drugs, anti-diabetics, sedatives, diuretics, COX-2 inhibitors, oxicams and Delta(9)-tetrahydro-11-norcannabinol-9-carboxylic acid.
Novelty and Efficiency of the Methods
- Placing particular emphasis on the unprecedented speed of these procedures, the researchers were able to achieve notably quick turnaround times. The total time required for both methods inclusive of post-run and equilibration times was 10 minutes.
- The methods utilize a high-efficiency reversed-phase column (3.3 cm L x 2.1 mm I.D. with 3 microm particles) which is paired with a fast-scanning triple-quadrupole mass spectrometer. The combination of these instruments results in fast and efficient screening.
Cite This Article
APA
Ho EN, Leung DK, Wan TS, Yu NH.
(2006).
Comprehensive screening of anabolic steroids, corticosteroids, and acidic drugs in horse urine by solid-phase extraction and liquid chromatography-mass spectrometry.
J Chromatogr A, 1120(1-2), 38-53.
https://doi.org/10.1016/j.chroma.2006.03.089 Publication
Researcher Affiliations
- Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, N.T., Hong Kong, PR China. emmie.nm.ho@hkjc.org.hk
MeSH Terms
- Adrenal Cortex Hormones / administration & dosage
- Adrenal Cortex Hormones / chemistry
- Adrenal Cortex Hormones / urine
- Anabolic Agents / administration & dosage
- Anabolic Agents / chemistry
- Anabolic Agents / urine
- Animals
- Chromatography, High Pressure Liquid / methods
- Doping in Sports
- Horses
- Molecular Structure
- Reproducibility of Results
- Spectrometry, Mass, Electrospray Ionization / methods
- Steroids / administration & dosage
- Steroids / chemistry
- Steroids / urine
- Substance Abuse Detection / methods
Citations
This article has been cited 9 times.- Yamini Y, Safari M, Shamsayei M. Simultaneous determination of steroid drugs in the ointment via magnetic solid phase extraction followed by HPLC-UV. J Pharm Anal 2018 Aug;8(4):250-257.
- Haneef J, Shaharyar M, Husain A, Rashid M, Mishra R, Parveen S, Ahmed N, Pal M, Kumar D. Application of LC-MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids. J Pharm Anal 2013 Oct;3(5):341-348.
- Ma Y, Gao S, Hu M. Quantitation of celecoxib and four of its metabolites in rat blood by UPLC-MS/MS clarifies their blood distribution patterns and provides more accurate pharmacokinetics profiles. J Chromatogr B Analyt Technol Biomed Life Sci 2015 Sep 15;1001:202-11.
- Rannulu NS, Cole RB. Novel fragmentation pathways of anionic adducts of steroids formed by electrospray anion attachment involving regioselective attachment, regiospecific decompositions, charge-induced pathways, and ion-dipole complex intermediates. J Am Soc Mass Spectrom 2012 Sep;23(9):1558-68.
- Goel R, Muthusamy B, Pandey A, Prasad TS. Human protein reference database and human proteinpedia as discovery resources for molecular biotechnology. Mol Biotechnol 2011 May;48(1):87-95.
- Wolf-Yadlin A, Hautaniemi S, Lauffenburger DA, White FM. Multiple reaction monitoring for robust quantitative proteomic analysis of cellular signaling networks. Proc Natl Acad Sci U S A 2007 Apr 3;104(14):5860-5.
- Karačonji IB, Jelača T, Jurič A, Vrdoljak AL. Morphine and codeine in racing horse feed: is there reason for concern?. Arh Hig Rada Toksikol 2025 Mar 1;76(1):60-66.
- Hajrulai-Musliu Z, Uzunov R, Jovanov S, Musliu D, Dimitrieska-Stojkovikj E, Stojanovska-Dimzoska B, Angeleska A, Stojkovski V, Sasanya JJ. Multi-class/residue method for determination of veterinary drug residues, mycotoxins and pesticide in urine using LC-MS/MS technique. BMC Vet Res 2023 Sep 14;19(1):156.
- Chiou SH, Wu CY. Clinical proteomics: current status, challenges, and future perspectives. Kaohsiung J Med Sci 2011 Jan;27(1):1-14.
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