Sonu Gandhi
Amity University, India
Title: A chemiluminescence approach for the detection of morphine in urine samples
Biography
Biography: Sonu Gandhi
Abstract
A sensitive immunoassay for morphine was developed by chemiluminescent approach. The anti-MAM antibody was developed for monoacetyl morphine (MAM) and labeled with horseradish peroxidase (HRP) for the development of competitive assay. The method is based on the competition between labeled antibodies and free drug in spiked urine samples. A hapten (MAM) was functionalized with chloroacetic acid chemistry to generate hydroxyl moiety and conjugated with carrier protein (BSA). The biophysical characterization was done by various spectroscopic methods such as UV-Vis, Fluorometer, CD, IR, MALDI-TOFF and was used as an immunogen for the generation of anti-MAM antibody. A binding assay was performed and high titer of antibody (1:64,0000) was obtained and the relative binding affinity constant (Kaff) of anti-MAM antibody was 3.1×107 l/mol. Under competitive conditions, the IC50 value for heroin, mono acetyl morphine (MAM), morphine and codeine were in the range of 0.01-0.08 ng/ml respectively. The developed chemiluminescence based competitive assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml that indicates fairly good precision with linear range, reactivity and therefore could be implement in illegal trafficking.