Semi-Quantitative On-Site Microfluidic Assay to Detect 11-Nor-9-carboxy-delta 9-Tetrahydrocannabinol (THC-COOH) in Urine

Abstract

The rapid detection of 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH), a primary cannabis metabolite, is critical for forensic and workplace drug testing. However, conventional immunoassays often lack sensitivity and objectivity. We developed a portable lateral flow immunoassay device with a microfluidic cartridge and fluorescent reader for the semi-quantitative detection of THC-COOH in urine. A test-to-reference fluorescence ratio was employed to mitigate matrix effects and ensure objective results. The device was validated for accuracy, repeatability, and stability using spiked urine samples and compared against validated LC-MS/MS results on 100 authentic urine samples (50 positive and 50 negative). At a cutoff of 20 ng/mL, the device achieved 100% sensitivity and specificity, with repeatability and reproducibility CVs of below 15%. The cutoff index (COI) strongly correlated with LC-MS/MS results (R2 = 0.9471). Crucially, this high correlation with hydrolyzed LC-MS/MS data demonstrates that the antibody recognizes both free and glucuronide-conjugated metabolites, validating its reliability without enzymatic pre-treatment. This microfluidic device enables rapid, sensitive on-site THC-COOH detection, featuring automated data management via Wi-Fi connectivity, enhancing its forensic applicability.