Rapid immunoassays for detection of anabolic nortestosterone in dietary supplements

Abstract

An enzyme immunoassay (ELISA) and an immunochromatographic strip were designed for a rapid detection of nortestosterone in dietary supplements. Two polyclonal antibodies and two types of nortestosterone-protein coating conjugates were tested to develop the most appropriate method. Under optimal experimental conditions, the most sensitive ELISA achieved the IC50 and the limit of detection values of 6.41 and 0.09 ng/ml, respectively. The assay specificity was tested measuring cross-reactivity of several steroids. The interference with the assay was negligible (< 0.1%), except for cross-reactivity with another frequently abused steroid testosterone (23%). The optimised gold particle-based immunochromatographic strip provided in semi-quantitative test a visual detection limit of 1 ng/ml. None of these methods showed the interference using a filtrate of the suspension of non-contaminated sample. After the validation for particular matrices, the ELISA and the strip test could be useful tools for a rapid analysis of nortesto-sterone in crude extracts of dietary supplements.