Description
Intended Use: The Flunitrazepam ELISA Kit is intended for the detection of Flunitrazepam in human serum.
Summary and Explaination: The Flunitrazepam Direct ELISA Kit is a sensitive in-vitro test to detect the presence of Flunitrazepam and 7-amino flunitrazepam in forensic samples such as whole blood, serum, plasma and urine.
Flunitrazepam (Rohypnol) is available in a number of western European countries and Mexico for use as a hypnotic and anesthetic induction agent. It is administered orally or by intravenous injection in does of 2 mg. Flunitrazepam is metabolized via N-demethylation, 3-hydroxylation and glucoronidation and the reduction of the nitro group to an amine with subsequent acetylation. Over a seven day period an average of 84% of a labeled dose is eliminated in urine and 11% in feces. Major metabolites include 7-aminoflunitrazepam, 3-hydroxyflunitrazepam, 7-acetamidonorflunitrazepam and 3-hydroxy-7acetamido-flunitrazepam.
Principle of the test: The Flunitrazepam Direct ELISA Kit is based upon the competitive binding to antibody of enzyme labeled antigen and unlabeled antigen, in proportion to their concentration in the reaction mixture.
A 20 ml. aliquot of a diluted unknown specimen is incubated with a 100 ml. dilution of enzyme (Horseradish peroxidase) labeled Flunitrazepam derivative in micro-plate wells, coated with fixed amounts of high affinity purified polyclonal anti-Flunitrazepam. The wells are washed thoroughly and a chromogenic substrate added. The color produced is stopped using a dilute acid stop solution and the wells read at 450 nm. The intensity of the color developed is inversely proportional to the concentration of drug in the sample. The technique is sensitive to 0.5 ng/ml.
The Flunitrazepam Direct ELISA Kit avoids extraction of urine or blood sample for measure--ment. It employs a Flunitrazepam directed antiserum. Due to the proprietary method of orienting the antibody on the polystyrene micro-plate much higher sensitivity is achieved compared to passive adsorption. This allows an extremely small sample size reducing matrix effects and interference with binding proteins(s) or other macro-molecules