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Biomarkers of alcohol abuse in racehorses by liquid chromatography/tandem mass spectrometry.
Rapid Commun Mass Spectrom. 2007; 21(23):3785-94.RC

Abstract

A rapid and sensitive method was developed for the screening, quantification and confirmation of ethyl glucuronide (EG) and ethyl sulfate (ES) as biomarkers for alcohol administration to racehorses using liquid chromatography coupled on-line with triple quadrupole tandem mass spectrometry. Urine sample aliquots (0.1 mL) were pre-treated by protein precipitation. Separation of EG and ES was achieved on an Ultra PFP column. Isocratic elution with a flush step was performed using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). Analysis was performed by negative electrospray ionization in multiple reaction monitoring mode. The retention times for EG and ES were 1.7 +/- 0.30 and 3.4 +/- 0.30 min, respectively. The internal standard used was d(5)-ethyl glucuronide with a retention time of 1.7 +/- 0.30 min. The entire separation was completed in <5 min. The limit of detection (LOD) and of quantification (LOQ) for both analytes were 100 ng/mL (S/N > or =3) and 500 ng/mL, respectively. The limit of confirmations (LOC) for EG and ES were 500 ng/mL and 1.0 microg/mL, respectively. The assay was linear over a concentration range of 0.5-100 microg/mL (r(2) > 0.995). Intra- and inter-day accuracy and precision were less than 15%. The analytes were stable in urine for 24 h at room temperature, 10 days at 4 degrees C and 21 days at -20 degrees C and -70 degrees C. Ion suppression or enhancement due to matrix effect was negligible. The measurement uncertainty was <14% for EG and <25% for ES. This method was successfully used for the quantification of EG and ES in urine samples following alcohol administration to research horses and for screening and confirmation of EG and ES in urine samples obtained from racehorses post-competition. The method is simple, rapid, inexpensive, and reliably reproducible.

Authors+Show Affiliations

You Y
University of Pennsylvania, School of Veterinary Medicine, Department of Clinical Studies, New Bolton Center Campus, Kennett Square, PA 19348, USA.
Uboh CE
No affiliation info available
Soma LR
No affiliation info available
Guan F
No affiliation info available
Li X
No affiliation info available
Rudy JA
No affiliation info available
Chen J
No affiliation info available

MeSH

AlcoholismAnimalsBiomarkersChromatography, High Pressure LiquidGlucuronatesHorsesReproducibility of ResultsSensitivity and SpecificitySpectrometry, Mass, Electrospray IonizationSubstance Abuse DetectionSulfuric Acid EstersUrinalysis

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17973234

Citation

You, Youwen, et al. "Biomarkers of Alcohol Abuse in Racehorses By Liquid Chromatography/tandem Mass Spectrometry." Rapid Communications in Mass Spectrometry : RCM, vol. 21, no. 23, 2007, pp. 3785-94.
You Y, Uboh CE, Soma LR, et al. Biomarkers of alcohol abuse in racehorses by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom. 2007;21(23):3785-94.
You, Y., Uboh, C. E., Soma, L. R., Guan, F., Li, X., Rudy, J. A., & Chen, J. (2007). Biomarkers of alcohol abuse in racehorses by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry : RCM, 21(23), 3785-94.
You Y, et al. Biomarkers of Alcohol Abuse in Racehorses By Liquid Chromatography/tandem Mass Spectrometry. Rapid Commun Mass Spectrom. 2007;21(23):3785-94. PubMed PMID: 17973234.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Biomarkers of alcohol abuse in racehorses by liquid chromatography/tandem mass spectrometry. AU - You,Youwen, AU - Uboh,Cornelius E, AU - Soma,Lawrence R, AU - Guan,Fuyu, AU - Li,Xiaoqing, AU - Rudy,Jeffrey A, AU - Chen,Jinwen, PY - 2007/11/2/pubmed PY - 2008/1/9/medline PY - 2007/11/2/entrez SP - 3785 EP - 94 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 21 IS - 23 N2 - A rapid and sensitive method was developed for the screening, quantification and confirmation of ethyl glucuronide (EG) and ethyl sulfate (ES) as biomarkers for alcohol administration to racehorses using liquid chromatography coupled on-line with triple quadrupole tandem mass spectrometry. Urine sample aliquots (0.1 mL) were pre-treated by protein precipitation. Separation of EG and ES was achieved on an Ultra PFP column. Isocratic elution with a flush step was performed using 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). Analysis was performed by negative electrospray ionization in multiple reaction monitoring mode. The retention times for EG and ES were 1.7 +/- 0.30 and 3.4 +/- 0.30 min, respectively. The internal standard used was d(5)-ethyl glucuronide with a retention time of 1.7 +/- 0.30 min. The entire separation was completed in <5 min. The limit of detection (LOD) and of quantification (LOQ) for both analytes were 100 ng/mL (S/N > or =3) and 500 ng/mL, respectively. The limit of confirmations (LOC) for EG and ES were 500 ng/mL and 1.0 microg/mL, respectively. The assay was linear over a concentration range of 0.5-100 microg/mL (r(2) > 0.995). Intra- and inter-day accuracy and precision were less than 15%. The analytes were stable in urine for 24 h at room temperature, 10 days at 4 degrees C and 21 days at -20 degrees C and -70 degrees C. Ion suppression or enhancement due to matrix effect was negligible. The measurement uncertainty was <14% for EG and <25% for ES. This method was successfully used for the quantification of EG and ES in urine samples following alcohol administration to research horses and for screening and confirmation of EG and ES in urine samples obtained from racehorses post-competition. The method is simple, rapid, inexpensive, and reliably reproducible. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/17973234/Biomarkers_of_alcohol_abuse_in_racehorses_by_liquid_chromatography/tandem_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -

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