Description
Intended Use: The Barbiturate Direct ELISA Kit provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/ mass spectrometry (GS-MS) is the preferred confirmatory method. Professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
Summary and Explaination: The Barbiturate Direct ELISA Kit is a specific and sensitive in-vitro test to detect the presence of barbiturates in samples such as whole blood, serum, plasma and urine. Barbiturates - derivatives of Barbituric acid - are sedative drugs which at low doses induces relaxation and at high doses induce coma and even death (2). Barbiturates are usually administered orally but may also be taken intravenously or intramuscularly and are absorbed rapidly. The metabolism of Barbiturates is mainly in the liver, a number of metabolic pathways have been described which include oxidation, desulfuration and ring cleavage. Because the number and the proportion of the various Barbiturate metabolites varies with each individual the results are expressed in terms of the equivalents of the standard, secobarbital/ml.
Principle of the test: The Barbiturate 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 10 ƒÝl. aliquot of a diluted unknown specimen is incubated with a 100 ƒÝl. dilution of enzyme (Horseradish peroxidase) labeled barbiturate derivative in micro-plate wells, coated with fixed amounts of oriented high affinity purified polyclonal antibody. 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 1 ng/ml. The Barbiturate Direct ELISA Kit avoids extraction of urine or blood sample for measure¬¬ment. It employs a polyclonal high affinity, purified Barbiturate antibody. Due to the proprietary method of orienting the antibody on the polystyrene micro-plate much higher sensitivity is achieved compared to passive adsorption. This results in extremely small sample size reducing matrix effects and interference with binding proteins(s) or other macro¬molecules