Clinical Chemistry
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Clinical Chemistry 45: 1224-1234, 1999;
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(Clinical Chemistry. 1999;45:1224-1234.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Broad Spectrum Drug Identification Directly from Urine, Using Liquid Chromatography-Tandem Mass Spectrometry

Robert L. Fitzgerald1,2,a, Jeffrey D. Rivera1 and David A. Herold1,2

1 Veterans Administration Medical Center San Diego and
2 University of California San Diego, 3350 La Jolla Village Dr., San Diego, CA 92161.
a Address correspondence to this author at: VAMC-113, 3350 La Jolla Village Dr., San Diego, CA 92161. Fax 619-552-7479; e-mail rlfitzgerald{at}vapop.ucsd.edu

Background: Currently the rate-limiting step for mass spectrometric analysis of drugs in biological samples is sample preparation. Many gas chromatography/mass spectrometry (GC/MS) methods are specific for a certain class of compounds, requiring extraction and/or derivatization before analysis. The purpose of this study was to develop a broad spectrum liquid chromatography/mass spectrometry (LC/MS) procedure that allowed for direct analysis of urine specimens with potential for quantitative analysis.

Methods: We modified a commercially available column-switching instrument, the REMEDi HS from Bio-Rad Diagnostics, to make it compatible with atmospheric pressure ionization. The system we developed was based on electrospray ionization and used three LC columns to extract, purify, and separate drugs directly from urine specimens. Drugs and metabolites were tentatively identified on the basis of retention times and (M+H)+ ions. Tandem mass spectrometry (MS/MS) was used to confirm the qualitative identification of suspected drugs, using data-dependent acquisition. For quantitative analysis, the cocaine metabolite benzoylecgonine was analyzed using isotope dilution and selected reaction monitoring.

Results: Seventeen basic drugs from a variety of classes of compounds were identified directly from urine without the need for prior sample extraction, using LC and MS/MS. Quantitative analysis was demonstrated for benzoylecgonine. When benzoylecgonine-d3 was used as the internal standard, the method was linear from 30 to 10 000 µg/L (range tested). At these concentrations, the within-run accuracy was ± 10% of the target concentration, with CVs <10%. Analytical results by LC/MS/MS compared favorably with GC/MS values for 50 benzoylecgonine-containing specimens and for 25 negative specimens.

Conclusions: The ability to directly analyze urine for a wide variety of drug classes, combined with the sensitivity and specificity of LC/MS/MS makes this technique attractive for many clinical, forensic, and biotechnology applications.© 1999 American Association for Clinical Chemistry




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