Rapid Analysis of Cocaine and Metabolites in Urine Using a Completely Automated Solid-Phase Extraction-High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method

Abstract

A rapid, simple, and completely automated method for the analysis of cocaine and its metabolites in urine has been developed. The method utilizes online solid-phase extraction (SPE) with liquid chromatographic separation and tandem mass spectrometric detection (MS-MS). An efficient online SPE procedure was developed using Hyspheretrade mark MM anion sorbent. A gradient chromatography method with a Gemini C6-Phenyl (50 x 3.00-mm i.d., 5 microm) column was used for the separation of all compounds. Detection was by positive ion mode electrospray ionization MS-MS. Multiple reaction monitoring (MRM) was used to enhance the selectivity and sensitivity of the method. Two MRM transitions were monitored for each analyte and one transition for each internal standard. Linearity was analyte-dependent but generally ranged from 7 to 1000 ng/mL. The limits of detection for the method ranged from 3 to 23 ng/mL and the limits of quantitation ranged from 7 to 69 ng/mL. Quality control (QC) samples were analyzed for each analyte in triplicate at 50, 200, and 800 ng/mL. The bias and precision were determined using a simple analysis of variance (ANOVA: single factor). The results demonstrated bias as < 5% and % precision as < 9% for all components at each QC level.

Assessing Cocaine Abuse Using LC–MS/MS Measurements in Biological Specimens

“…LC-MS/MS (ESI) 23 ng/ml; 69 ng/ml (ECG) [63] 11 ng/ml; 25 ng/ml (EME) 4 ng/ml; 13 ng/ml (BE) 2 ng/ml; 7 ng/ml (COC) 7 ng/ml; 21 ng/ml (COET) -tetrahydrocannabinol-9-carboxylic acid.…”

“…Using only 0.1 ml of sample, quantitation limits of 15 ng/ml for cocaine and metabolites were obtained, and recoveries were higher than 85% for these compounds. According to the authors, this fully automated procedure allowed reducing the sam- LC-MS/MS (ESI) 10 ng/ml; 5 ng/ml (EME) [52] 2 ng/ml; 8 ng/ml (BE) 10 ng/ml; 8 ng/ml (COC) 2 ng/ml; 8 ng/ml (NCOC) 2 ng/ml; 7 ng/ml (COET) Assessing cocaine abuse using LC-MS/MS measurements in biological specimens Review [73] 0.9 pg/mg; 3.0 pg/mg (NCOC) [63], yet using higher amounts of sample and comparable detection limits. Another interesting approach is the use of molecularly imprinted polymers for the selective analysis of cocaine and metabolites from urine samples [36].…”

Optimization and validation of simultaneous analyses of ecgonine, cocaine, and seven metabolites in human urine by gas chromatography–mass spectrometry using a one‐step solid‐phase extraction

“…Only a few methods, based on gas chromatography–mass spectrometry (GC–MS) or liquid chromatography–mass spectrometry tandem (LC–MS/MS), have been published for the determination of ECG, COC, and other metabolites in biological fluids.…”

Comprehensive automation of the solid phase extraction gas chromatographic mass spectrometric analysis (SPE-GC/MS) of opioids, cocaine, and metabolites from serum and other matrices

“…The compounds were analyzed from different matrices like urine [9, 28, 29], whole blood, serum, plasma [5, 9, 7], saliva [6], hair [8], or post-mortem samples [17]. In this study, a completely automated SPE-GC/MS method was developed from a validated, partly automated (SPE) analysis method.…”