Biomaterial Database

Doping control analysis of four JWH-250 metabolites in equine urine by liquid chromatography-tandem mass spectrometry. 2019

Youwen You, and Rachel M Proctor, and Eric D Vasilko, and Mary A Robinson

School of Veterinary Medicine, Department of Clinical Studies, University of Pennsylvania, New Bolton Center Campus, Kennett Square, Pennsylvania, 19348, USA.

JWH-250 is a synthetic cannabinoid. Its use is prohibited in equine sport according to the Association of Racing Commissioners International (ARCI) and the Fédération Équestre Internationale (FEI). A doping control method to confirm the presence of four JWH-250 metabolites (JWH-250 4-OH-pentyl, JWH-250 5-OH-pentyl, JWH-250 5-OH-indole, and JWH-250 N-pentanoic acid) in equine urine was developed and validated. Urine samples were treated with acetonitrile and evaporated to concentrate the analytes prior to the analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The chromatographic separation was carried out using a Phenomenex Lux® 3 μm AMP column (150 x 3.0 mm). A triple quadrupole mass spectrometer was used for detection of the analytes in positive mode electrospray ionization using multiple reaction monitoring (MRM). The limits of detection, quantification, and confirmation for these metabolites were 25, 50, and 50 pg/mL, respectively. The linear dynamic range of quantification was 50-10000 pg/mL. Enzymatic hydrolysis indicated that JWH-250 4-OH-pentyl, JWH-250 5-OH-pentyl, and JWH-250 5-OH indole are highly conjugated whereas JWH-250 N-pentanoic acid is not conjugated. Relative retention time and product ion intensity ratios were employed as the criteria to confirm the presence of these metabolites in equine urine. The method was successfully applied to post-race urine samples collected from horses suspected of being exposed to JWH-250. All four JWH-250 metabolites were confirmed in these samples, demonstrating the method applicability for equine doping control analysis.

UI	MeSH Term	Description	Entries
D007211	Indoles	Benzopyrroles with the nitrogen at the number one carbon adjacent to the benzyl portion, in contrast to ISOINDOLES which have the nitrogen away from the six-membered ring.
D002186	Cannabinoids	Compounds having the cannabinoid structure. They were originally extracted from Cannabis sativa L. The most pharmacologically active constituents are TETRAHYDROCANNABINOL; CANNABINOL; and CANNABIDIOL.	Cannabinoid
D002853	Chromatography, Liquid	Chromatographic techniques in which the mobile phase is a liquid.	Liquid Chromatography
D006736	Horses	Large, hoofed mammals of the family EQUIDAE. Horses are active day and night with most of the day spent seeking and consuming food. Feeding peaks occur in the early morning and late afternoon, and there are several daily periods of rest.	Equus caballus,Equus przewalskii,Horse, Domestic,Domestic Horse,Domestic Horses,Horse,Horses, Domestic
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D000840	Anisoles	A group of compounds that are derivatives of methoxybenzene and contain the general formula R-C7H7O.	Methylphenyl Ethers,Ethers, Methylphenyl
D015813	Substance Abuse Detection	Detection of drugs that have been abused, overused, or misused, including legal and illegal drugs. Urine screening is the usual method of detection.	Drug Abuse Testing,Illicit Drug Testing,Street Drug Testing,Substance Abuse Testing,Drug Abuse Detection,Drug Abuse Screening,Illicit Drug Detection,Street Drug Detection,Detection, Drug Abuse,Detection, Illicit Drug,Detection, Street Drug,Detection, Substance Abuse,Detections, Drug Abuse,Detections, Illicit Drug,Detections, Street Drug,Detections, Substance Abuse,Drug Abuse Detections,Drug Abuse Screenings,Drug Abuse Testings,Drug Testing, Illicit,Drug Testings, Illicit,Illicit Drug Detections,Illicit Drug Testings,Screening, Drug Abuse,Screenings, Drug Abuse,Street Drug Detections,Street Drug Testings,Substance Abuse Detections,Substance Abuse Testings,Testing, Drug Abuse,Testing, Illicit Drug,Testing, Street Drug,Testing, Substance Abuse,Testings, Drug Abuse,Testings, Illicit Drug,Testings, Street Drug,Testings, Substance Abuse
D053719	Tandem Mass Spectrometry	A mass spectrometry technique using two (MS/MS) or more mass analyzers. With two in tandem, the precursor ions are mass-selected by a first mass analyzer, and focused into a collision region where they are then fragmented into product ions which are then characterized by a second mass analyzer. A variety of techniques are used to separate the compounds, ionize them, and introduce them to the first mass analyzer. For example, for in GC-MS/MS, GAS CHROMATOGRAPHY-MASS SPECTROMETRY is involved in separating relatively small compounds by GAS CHROMATOGRAPHY prior to injecting them into an ionization chamber for the mass selection.	Mass Spectrometry-Mass Spectrometry,Mass Spectrometry Mass Spectrometry,Mass Spectrometry, Tandem
D057230	Limit of Detection	Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure.	Limits of Detection,Detection Limit,Detection Limits