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Determination of Acepromazine and its Major Metabolite in Equine Serum by LC-MS/MS using the Finnigan LCQ Deca XP Plus Ion Trap Mass Spectrometer

Chromatography and Mass Spectrometry Application Report

Key Words Sensitivity Quantitation Finnigan LCQ Deca XP Plus Equine serum Acepromazine

Gargi Choudhary, Thermo Electron Corporation, San Jose, CA USA
Wayne Skinner and Scott Stanley, University of California, Davis

Introduction

Acepromazine is widely used as a sedative in horses. It serves as a tranquilizer to better facilitate handling of these large mammals before transportation or surgical procedures. Acepromazine has been classified by the Association of Racing Commissioners International, Inc. as a Class 3 drug in horses. Abuse of the drugs in the Class 3 category can result in 2-6 months of suspension, up to $1500 fine, and loss of purse. Acepromazine has a slow elimination rate and stays in the body for a long time. This can be problematic if the drug is administered before a race event since its calming effect can lead to a better performance by the horse.

Goal

Develop a method for identifying and quantitating acepromazine (ACE) and its major metabolite 2-(1-hydroxyethyl) promazine sulfoxide (HEPS) in horse serum.
Determine limits of detection (LOD) and quantitation (LOQ) of these two compounds in serum.
Demonstrate low level detection of acepromazine in a complex biological matrix.

Experimental

HPLC
HPLC system: Surveyor MS pump with Surveyor autosampler
Column: 50 2.1 mm Hypersil BDS C18 (Thermo Electron)
Injection volume: 20 μL
Flow rate: 300 μL/min
Mobile phase A: Water containing 10 mM ammonium acetate and 0.1% formic acid
B: Acetonitrile containing 0.1% formic acid
Gradient: 5-40% B in 4 min, 40-65% B in 1
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