Confirmation of Marijuana, Cocaine, Morphine, Codeine, Amphetamine, Methamphetamine, Phencyclidine by GC/MS in Urine Following Immunoassay Screening

Abstract

Rapid, reliable, sensitive, qualitative, and quantitative methods using small urine volumes (0.2–0.5 mL) were developed primarily for confirmation of marijuana, cocaine, benzoylecgonine, ecgonine methyl ester, morphine, codeine, amphetamine, methamphetamine, and phencyclidine. Using capillary gas chromatography/mass spectrometry (GC/MS) and selected ion monitoring (SIM), mass spectra were obtained for each analyte. Samples were prepared by hydrolysis where applicable, organic solvent extraction, and derivatization where necessary. Confirmation was achieved by comparing abundance of major ions and retention time of the total ion current (TIC) of an analyte with those of the appropriate analytical standard. Quantitation was achieved and calibration curves derived by obtaining the molecular ion ratios of that analyte/internal standard (IS) over a concentration range of 10–300 ng/mL (0.16–4.0 ng total injected into GC/MS). The overall extraction efficiency for these analytes ranged from 53% to 96%. Statistically significant cut-off values (p < 0.01) were obtained for each analyte. The slope, y-intercept, and coefficient of determination (r²) were calculated for each analyte. All of the GC/MS methods were extensively tested against urine samples determined positive or negative by immunoassay (IA) and are now used in our laboratory.