Abstract
BACKGROUND: Oral fluid is a widely studied matrix able to isolate the primary Cannabis constituent THC, facilitating its detection via mass spectrometry, and in most cases link these findings to recent drug use. As an alternative to liquid oral fluid, dried oral fluid spots (DOFS) is a simple and a low-cost sampling technique. It has shown improved stability compared to liquid samples, allowing for the possibility to preserve the specimens under various temperature and humidity conditions. The sampling strategy is straightforward and involves the application of a small quantity of oral fluid aliquot to a paper substrate that is set to air dry allowing for on-site collection at a large-scale demand. The goal of this study is to study THC and CBD extraction from DOFS, applying a previous established protocol for a LC-MS/MS qualitative method validation. Although other drugs of abuse have been included in DOFS methods, this is the first method validation including cannabinoids. An alternative oral fluid extraction method (WAX-S tips) is demonstrated to improve the recovery of the analytes. METHODS: A pool of blank oral fluid was used to prepare THC and CBD spiked DOFS samples for method validation and application. Spiked oral fluid was used to demonstrate WAX-S tips THC and CBD extraction. All samples were analyzed on a LC-MS/MS instrument. RESULTS: The qualitative method validation for THC and CBD confirmation in DOFS included method selectivity, matrix effects (< 20%), recovery (average of 25%), process efficiency (average of 21%), LOD (2 ng/mL for THC and 4 ng/mL for CBD), absence of carryover, and DOFS stability (70% in 35 days) as figures of merit. The method application in blindly prepared samples demonstrated the method capability to identify THC and CBD. WAX-S tips extraction showed an average of 91% recovery of THC and CBD from liquid oral fluid. CONCLUSIONS: THC and CBD extraction from DOFS showed low recoveries. However, the LC-MS/MS qualitative confirmation of THC and CBD in DOFS could improve cannabinoids screening in oral fluid, as it shows adequate LOD and stability over time. This method has potential for assisting the screening of drivers under possible drug influence by facilitating sample transportation and temporary storage in dried spot form. Additional research is suggested for WAX-S tips extraction and quantitative method validation.