Detection of 4 Benzodiazepines in Oral Fluid as Biomarker for Presence in Blood

Abstract

Analysis of samples of oral fluid (mixed saliva) is increasingly being used to detect recent drug use. The aim of this investigation was to assess the suitability of testing oral fluid as a biomarker for the presence of 4 benzodiazepines in blood and its possible application in clinical settings and in research on drug use. Paired samples of oral fluid and blood from 4080 individuals in 4 European countries were collected and analyzed for benzodiazepines using gas or liquid chromatography with mass spectroscopic detection. Concentration data for the 4 most commonly detected benzodiazepines were studied: alprazolam, clonazepam, diazepam, and nordiazepam. Large variations in oral fluid to blood concentration ratios were observed for the studied benzodiazepines. The interquartile ranges for the oral fluid to blood concentrations ratios corresponded to 88%-197% of the median values. Selecting cutoff concentrations in oral fluid that gave the best accuracy in identifying individuals with benzodiazepine concentrations in blood above chosen thresholds produced accuracies of 74%-85% and the fraction of false negatives was 9%-23%. The concentration of the 4 studied benzodiazepines in oral fluid can neither be used to accurately estimate the concentrations in blood nor to correctly identify patients with blood drug concentrations below or above recommended therapeutic levels. When using analytical methods with limits of quantitation corresponding to concentrations less than 0.5 ng/mL in undiluted oral fluid, it may be used to confirm a recent intake of benzodiazepines. However, it is likely that some false negatives may occur.