Validation of a Novel Immunoassay for the Detection of Synthetic Cannabinoids and Metabolites in Urine Specimens

Abstract

Synthetic cannabinoid drugs do not cross react on traditional marijuana immunoassay tests, preventing their use in large scale drug screening programs. This paper describes the validation and performance characteristics of two enzyme linked immunosorbent assays designed to detect the use of two common synthetic cannabinoids in urine, JWH-018 and JWH-250. The JWH-018 assay has significant cross-reactivity with several synthetic cannabinoids and their metabolites, whereas the JWH-250 assay has limited cross-reactivity. The assays are calibrated at 5 ng/mL with the 5-OH metabolite of JWH-018 and the 4-OH metabolite of JWH-250. The method was validated with 114 urine samples for JHW-018 and 84 urine samples for JWH-250 and confirmed by using liquid chromatography tandem mass spectrometry, which tests for metabolites of JWH-018, JWH-019, JWH-073, JWH-250 and AM-2201. The accuracy was determined to be 98% with greater than 95% sensitivity and specificity for both assays.

Simultaneous quantification of 37 synthetic cannabinoid metabolites in human urine by liquid chromatography-tandem mass spectrometry

“…However, synthetic cannabinoids cannot be detected by most preliminary immunoassay screening procedures, which contribute to the prevalent use of these drugs. In spite of the recent development of immunoassays that target synthetic cannabinoids, they are still limited due to insufficient specificity and the continuous emergence of newly modified synthetic cannabinoids [13,14]. Thus, some screening methods employing mass spectrometric techniques have been developed for selective and sensitive detection of synthetic cannabinoids and their metabolites in various biological matrices [15][16][17][18][19].…”

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

“…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

“…In SCs extraction, LLE is usually used because of their high hydrophobicity. It has been used for the extraction of SCs in the following matrices: blood [35,[66][67][68][69][70][71][72], serum [39,[73][74][75][76], plasma [77], urine [40,41,47,49,51,59,72,[78][79][80][81][82][83], oral fluid [84,85] and hair [86][87][88][89][90]. It involves adding an immiscible organic solvent such as tert-butyl methyl ether [41,82], chloroform [50] and diethyl ether [40,52,79] to the sample, and mixing, removing the organic solvent.…”

“…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]. …”

“…In SCs' analysis, positive polarity is commonly performed. The negative ionisation mode has been used for HU-210, CP 47,497 and its homologue due to the absence of nitrogen in the chemical structures of these compounds in contrast to almost all other SCs [82].…”