Homologous ELISA for detection of 17α-methyltestosterone in serum

17 α Methyl testosterone is a synthetic steroid hormone having androgenic properties. It used in aquaculture, medical treatments, sports as doping drugs, improving food efficacy but its residue meat and their products can cause risk to the human, animals environment. In this article, we have focused on developing homologous ELISA using 17α testosterone-3-Carboxymethyloxime-Bovine serum albumin antiserum testosterone-3-Carboxymethyloxime-Horseradish peroxidase enzyme conjugate. Immunogen conjugate both were prepared by N-hydroxysuccinimide mediated carbodiimide reaction method. The sensitivity, affinity ED50 of assay found be 0.11 ng/ml, 0.02x10-8 L/mol 5.78 ng/ml respectively. cross reactivity for combination was seen with eight steroids out 59 structurally related steroids. then studied analytical variables like recovery (94.8%-111.6%), precision (Inter Intra-assay coefficient variation less than 10%), correlation (R2= 0.96) comparing commercially available kit further validated measuring levels methyltestosterone rat after injecting rats intramuscularly.