Detection of anti-diabetics in equine plasma and urine by liquid chromatography-tandem mass spectrometry.
OBJECTIVE
Anti-diabetics such as sulfonylurea and thiazolidinedione derivatives are hypoglycemic drugs used for the treatment of diabetes. However, they can also be used as a stopper in horseracing. This paper describes a convenient method for the separation and simultaneous detection of 10 anti-diabetic drugs (namely glipizide, glibenclamide, glimepiride, gliclazide, tolazamide, tolbutamide, nateglinide, repaglinide, rosiglitazone and pioglitazone) in equine plasma and urine by LC-MS-MS.


METHODS
The anti-diabetics were isolated from equine plasma and urine by liquid-liquid extraction with 1,2-dichloroethane at acidic pH, and analysed by LC-MS-MS in the positive electrospray ionisation mode. Separation of 10 anti-diabetic drugs was achieved with a reversed phase C8 column using a mixture of aqueous ammonium formate (pH 3.0, 10 mM) and methanol as the mobile phase.


RESULTS
Detection and confirmation of the 10 anti-diabetic drugs at 10 ng/mL each in equine plasma and equine urine were achieved by full-scan MS-MS. All of these drugs were detected consistently at this concentration in spiked samples of different plasma and urine (n = 15 each). No significant matrix interferences were observed at the expected retention times of the targeted ions in blank urine samples (n = 30). This method has been used successfully in the analysis of drug-administration samples as well as official racing samples.


CONCLUSIONS
An LC-MS-MS method has been developed for the simultaneous detection of 10 anti-diabetics in equine plasma and urine. This method can be used to detect the abuse of anti-diabetic drugs in racehorses.
Racing Laboratory, The Hong Kong Jockey Club, Sha Tin Racecourse, Sha Tin, NT, Hong Kong, China. emmie.nm.ho@hkjc.org.hk
Emmie N M
EN
Ho
Kenneth C H
KC
Yiu
Terence S M
TS
Wan
Brian D
BD
Stewart
Keith L
KL
Watkins
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007004:Hypoglycemic Agents / Q000097:blood* / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013552:Swine
D007004:Hypoglycemic Agents