In recent years, the undeclared presence of various anabolic androgenic steroids (AAS) in commercial supplements has been confirmed. This fact can be a potential threat to all athletes using these supplements, and therefore, there is of increased interest in the implementation of rapid methods for the detection of AAS. The presented study describes the development of an immunostrip test for the detection of multiple 17α-methylated AAS based on direct and indirect competitive principle using gold nanoparticles as a label. As a capture reagent on test lines conjugated stanazolol to rabbit serum albumin (RSA/ST-3) was used, the intensity of color formed in the test line of the AAS-positive sample was visually distinguishable from that of negative sample within 10 min. The optimized closed direct and indirect format of the test provided a similar visual detection limit (0.7 and 0.9 ng/mL, respectively). The most commonly orally abused AAS (17α-methyltestosterone, methandienone, methyldihydrotestosterone, oxandrolone and oxymetholone) showed a strong cross-reaction. Developed immunostrips were successfully applied to analysis of artificially contaminated dietary supplements with 17α-methylated AASs. The developed immunostrips offer potential as a useful user-friendly method for capturing suspicious dietary supplement samples with different contents of AAS at levels far below the usually used concentrations of AAS.

• Keywords
• 17α-methylated AAS, anabolic steroids (AAS), dietary supplements, immunochromatography, stanazolol,
• Publication type
• Journal Article MeSH

In recent years, the use of synthetic cannabinoids (SCs) as drugs of abuse has greatly increased. SCs are associated with a risk of severe poisoning or even death. Therefore, more rapid, cost effective and reliable methods are needed, especially for the screening of drivers after traffic accidents and for detailed toxicological analysis in forensic laboratories. In this study, we developed a lateral flow immunoassay (LFIA) and an enzyme linked immunosorbent assay (ELISA) for the detection of JWH-200 in oral fluids. For this purpose a new hapten was prepared using a ten-step synthetic route. The developed immuno methods are based on antibodies obtained from rabbit immunized with synthesized hapten conjugated to carrier protein. The proposed methods are highly sensitive (LODLFIA = 0.08 ± 0.04 ng mL-1; LODELISA = 0.04 ± 0.02 ng mL-1). They were applied to the quantification of JHW-200 in spiked oral fluids. The recoveries ranged from 82 to 134% for both methods. The results correlated excellently with results obtained using UHPLC-MS/MS (R2LFIA = 0.99; R2ELISA = 0.99). Our developed methods could be an important tool for analyses of JWH-200 in human oral fluids. The one-step LFIA is particularly suitable for roadside and on-site monitoring due to the rapid qualitative results it delivers, while the ELISA is especially useful for laboratory quantitative analyses of positive samples captured by LFIA.

• Keywords
• BSA, bovine serum albumin, DCC, N,N’-dicyclohexylcarbodiimide, DIBAH, diisobutylaluminium hydride, DMF, N,N-dimethylformamide, ELISA, ELISA, enzyme-linked immunosorbent assay, GAR, goat anti-rabbit antibody, GAR-Po, peroxidase labelled goat anti-rabbit antibody, Hapten synthesis, Immunomethods, JWH-200, LFIA, LFIA, lateral flow immunoassay, LOD, limit of detection, NBS, N-bromosuccinimide, NHS, N-hydroxysuccinimide, NPS, new psychoactive substances, PEG, polyethylene glycol, RSA, rabbit serum albumin, RSD, relative standard deviation, SCs, synthetic cannabinoids, Synthetic cannabinoid, THC, thin layer chromatography,
• Publication type
• Journal Article MeSH