Validation of an LC-MS/MS Method for Analysis of 58 Drugs of Abuse in Oral Fluid and Method Comparison with an Established LC-HRMS Method.

Abstract

Liquid chromatography-mass spectrometry (LC-MS) methods for detection of multiple drugs of abuse (DoA) in oral fluid (OF) samples are being implemented in many clinical routine laboratories. Therefore, there is a need to develop new multi-analyte methods with simple sample pre-treatment and short analysis times. The purpose of this work was to validate a method detecting 58 DoA to be used with two different OF sampling kits, the saliva collection system (SCS) from Greiner Bio-One and Quantisal from Immunalysis, using the same sample pretreatment and analytical method. A set of 110 samples collected with the SCS kit was further compared to an LC-HRMS (high resolution mass spectrometry) method in another laboratory. The method was successfully validated, with precision and accuracy of ≤15% and z-scores of <2 for external controls. Using a sensitive LC-MS/MS instrument, the detection limits were <1 µg/L in neat oral fluid. In the comparative study between the LC-MS/MS and LC-HRMS methods using SCS samples, a good agreement was observed. Discrepancies were limited to lower concentration ranges, attributable to differences in cut-off thresholds between the methods. This work contributes to the development of LC-MS multi-analyte methods for OF samples, which are suitable for clinical routine laboratories.