Rapid and simple LC–MS/MS determination of urinary ethyl glucuronide, naltrexone, 6β-naltrexol, chlordiazepoxide, and norchlordiazepoxide for monitoring alcohol abuse

Abstract In this study, a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to detect ethyl glucuronide (EtG), which is a biomarker for monitoring alcohol consumption, and naltrexone (NTX), 6β-naltrexol (6βNTX), chlordiazepoxide (CDP), and norchlordiazepoxide (norCDP), which are analyzed to confirm the presence of medications for alcohol dependence treatment. The protein precipitation method was conducted to rapidly prepare samples. LC–MS/MS analysis was performed in the multiple-reaction monitoring mode. The analytes were separated using a Scherzo SM-C18 (2.0 × 100 mm, 3 µm) column. The calibration ranges were 5–1000 ng/mL for EtG, 6βNTX, CDP, and norCDP, and 1–100 ng/mL for NTX, with the correlation coefficients (r) being ≥ 0.994, and the weighting factor being 1/x 2. The lower limit of quantification was 1–5 ng/mL. The method was also validated for precision, accuracy, selectivity, dilution integrity, recovery, matrix effect, and stability. The developed method was successfully applied for the determination of EtG, NTX, 6βNTX, CDP, and norCDP in urine samples obtained from 49 probationers who received alcohol dependence treatment orders. The method developed herein can be used to monitor the drug-based treatment of alcohol abuse and alcohol consumption during the treatment of individuals under probation.