Evaluation of a Solid-Phase Extraction Method for Benzoylecgonine Urine Analysis in a High-Throughput Forensic Urine Drug-Testing Laboratory

Abstract

A novel extraction and derivatization procedure for the cocaine metabolite benzoylecgonine (BZE) was developed and evaluated for use in a high-volume forensic urine analysis laboratory. Extractions utilized a Speedisk 48 positive pressure extraction manifold and polymer-based cation-exchange extraction columns. Samples were derivatized by the addition of pentafluoropropionic anhydride and pentafluoropropanol. All analyses were performed in selected ion monitoring mode; ions included m/z 421, 300, 272, 429, and 303 with m/z 421 to 429 ratio used for quantitation. The average extraction efficiency was 80%. Seventy-five common over-the-counter products, including prescription drugs, drug metabolites, and other drugs of abuse, demonstrated no significant interference with respect to chromatography or quantitation. The limit of detection and limit of quantitation were calculated at 12.5 ng/mL, and the assay was linear from 12.5 to 20,000 ng/mL with an r2 of 0.99932. A series of 20 precision samples (100 ng/mL) produced an average response of 97.8 ng/mL and a percent coefficient of variation of 4.1%. A set of 79 archived human urine samples that had previously been found to contain BZE were analyzed by 3 separate laboratories. The results did not differ significantly from prior quantitation or between laboratories. The Speedisk has proven viable for a high-volume production facility reducing overall cost of analysis by decreasing analysis time and minimizing waste production while meeting strict forensic requirements.

Determination of cocaine, its metabolites, pyrolysis products, and ethanol adducts in postmortem fluids and tissues using Zymark® automated solid-phase extraction and gas chromatography-mass spectrometry1

“…TMS was also found to be an unstable derivative for the analysis of the COC-related analytes. The use of PFP/PFPA as a derivatizing agent provided a more stable derivative [20,31] with a significantly higher molecular weight and less background noise than TMS, which is generally used for COC-related analyses. PFPA derivatizes hydroxyl and secondary amine functional groups contained within many of the COC-related compounds.…”

Forensic Science

“…Urine analysis of laboratory personnel preparing cocaine-training aids for dogs was performed using GC/MS (313). Cocaine and its metabolites have been analyzed in biological fluids using liquidliquid extraction and GC (314), GC/MS (315,316), LC/MS/MS (317), HPLC/MS (318,319), HPLC (320), LC/APIMS (321), SPE and GC/MS (322), GC/FID and HPLC/UV (323), HPTLC/UV (324), solvent microextraction and GC (325), and HFMSME/GC (326). The detection of cocaine and metabolites in oral fluids has been accomplished by EIA and GC/MS (327) and HFMSME and GC (328).…”

Occupational Cocaine Exposure of Crime Laboratory Personnel Preparing Training Aids for a Military Working Dog Program

“…Benzoylecgonine was extracted using a Speedisk 48 TM positive-pressure extraction manifold (San Pedro, CA) as previously described (7). The limit of detection (LOD) was determined to be 12 ng/mL using a 2-mL urine aliquot.…”