Detection of Synthetic Cannabinoids in Oral Fluid Using ELISA and LC-MS-MS

Abstract

Synthetic cannabinoids are often referred to as 'Spice' or K2 compounds. Detection of these compounds in oral fluid has, to date, been limited to chromatographic procedures such as liquid chromatography with tandem mass spectrometry detection. We report the first analytical immunoassay for the screening of some synthetic cannabinoids in oral fluid specimens collected with the Quantisal™ device. JWH-200 was chosen as the calibration standard, because parent compounds, not metabolites, are predominantly detected in oral fluid. The immunoassay is capable of detecting JWH-200, JWH-018, JWH-073, JWH-022, AM-2201, AM-2232 and AM-1220. The assay was validated according to accepted laboratory protocols and applied to 32 authentic oral fluid specimens previously analyzed using LC-MS-MS at an accredited laboratory. The assay is sensitive, with a cutoff concentration of 0.25 ng/mL, and has a wide working range from 0.1 to 5 ng/mL. Intra- and interday precision were determined to be <10%. The screening method was completely validated and characterized; critical aspects of the screening included the incorporation of a preincubation step that improves the sensitivity of the assay to allow relevant concentrations of synthetic compounds in oral fluid to be detected.

Method Validation of The Biochip Array Technology for Synthetic Cannabinoids Detection in Urine

“…Cutoff optimization improved performance to 87.6% sensitivity, 85.2% specificity, and 85.4% efficiency. Conclusion: This highthroughput urine SC assay has good sensitivity and improved specificity and efficiency at modified cutoff concentrations.Bioanalysis (2014) 6(21) future science group Research Article Castaneto, Desrosiers, Ellefsen et al currently no regulated SC screening and confirmatory cutoffs, but SC markers were documented in hair [16], ante- [17,18] and postmortem blood [19], oral fluid [20][21][22], and urine [23][24][25][26][27] primarily by LC-MS/MS. Commercially available immunoassays also can provide rapid screening identification and semi-quantitative results for SC and metabolites, but critical assay evaluations are limited [22,28].…”

“…Commercially available immunoassays also can provide rapid screening identification and semi-quantitative results for SC and metabolites, but critical assay evaluations are limited [22,28].…”

Bioanalytical methods for the determination of synthetic cannabinoids and metabolites in biological specimens

“…Intercept [85] and Quantisal TM [97,101,124]. Kneisel et al [84] They also observed some degradation for JWH-073 and JWH-018 when stored at room temperature for one week, while when refrigerated, samples were stable.…”

“…Moreover, cross-reactivity and false positives can occur, and as such, toxicologists recommend that all positive immunoassay findings must be retested and confirmed by chromatographic analysis. Immunoassay tests have been used for the screening of SCs in oral fluid [101] and their metabolites in urine samples [82,93,[102][103][104]. …”

“…It is easy and rapid to obtain and is less prone to adulteration [84]. In addition, oral fluid containing parent SCs make the sample a good choice for testing in cases where metabolite references may be lacking, or if the target metabolites are still unknown [101]. However, there are some disadvantages; the sample volume must be sufficient for the analysis, and this amount could be difficult to obtain from some SCs abuser.…”