Published: Journal of Analytical Toxicology, ISSN 0146-4760, Volume 27, Number 6, September, pp. 325-331

Large-Volume Injection Gas Chromatography– Mass Spectrometry for Automated Broad-Spectrum Drug Screening in Horse Urine
Scott D. Stanley, Daniel McKemie, and Wayne Skinner
University of California, Davis, California Animal Health and Food Safety Laboratory, Kenneth L. Maddy Equine Analytical Chemistry Laboratory, West Health Science Drive, Davis, California 95616

A rapid, sensitive, and rugged method for detecting drugs and drug metabolites in extracts of horse urine is described. The use of large-volume injection (LVI) gas chromatography–mass spectrometry (GC–MS) for analysis of horse urine extracts allowed automation of the derivatization procedure and reduction of the sample volume from 5 mL to 1 mL of urine. An autosampler and temperature-programmable inlet were used to automatically dissolve the sample extract and form trimethylsilyl derivatives of over 200 analytes. The suitability of this procedure for routine GC–MS detection of approximately 80 basic analytes in extracts of racehorse urine was investigated. The formation of derivatives using LVI with in-liner derivatization was compared to a manual procedure involving the dissolution of sample extracts in N,O-bis(trimethylsily)trifluoroacetamide, heating the resulting mixture, and injecting 1 or 2 µL of the mixture through a splitless injector into the GC–MS instrument. In all cases, the in-liner derivatization reactions were found to be as complete as conventional heating block procedures. Ruggedness testing of the method demonstrated that peak resolution, shape, and area were maintained through 40 consecutive injections of sample extracts. No evidence of the accumulation of interfering substances was observed. The limits of detection using LVI GC–MS for routine screening of basic drugs in urine were generally in the range of 5–25 ng/mL. The method is currently being used to detect basic analytes in horse urine extracts with a throughput of approximately 50 urine sample extracts per instrument per day.

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