Cocaine Metabolite ELISA

Category Name

Drugs of Abuse

Method

ELISA

Principle

ELISA

Controls Standards

0,10,25,50ng/ml

Sensitivity

1ng/ml

Sample

10 µl

Runtime

90 min

Wavelength

450 nm

Safety

Please refer to MSDS

Shelflife

18 Months

CO084D-R5

Cocaine Metabolite MSDS

INTENDED USE

The Calbiotech, Inc. (CBI) Cocaine Metabolite Direct ELISA Kit is a specific and sensitive in-vitro test to detect the presence of Benzoylecgonine (BE) in forensic samples such as whole blood, serum, plasma and urine.

Summary AND EXPLANTION

Cocaine abuse is widespread and its prevalence may be increasing in all social and age strata (1). The drug is generally inhaled or smoked (1,2). Several methods for measurement of Cocaine Metabolite in urine exist (3-6). Benzoylecgonine, a major metabolite appears within minutes in urine (3). Since the number and proportion of metabolites vary in subjects, results are expressed in benzoylecgonine equivalents per ml. The Cocaine Metabolite Direct ELISA Kit is a single incubation assay providing results similar to those obtained by existing methods (4-6). Native (unaltered) cocaine urine concentration is far lower than that of its major metabolite benzoylecgonine. After intra- venous administration of l00mg cocaine urine concentrations ranged from 1.2 - 2.4 ug/ml compared with concentrations ranging from 5 - 55 ug/ml for benzoylecgonine (3). Cocaine was undetectable (at a 50 ng/ml cut-off) 12 hours after administration in comparison with benzoylecgonine which persists hours to days after administration (7). It has been suggested that a benzoylecgonine/cocaine ratio of less than 100 is indicative of use within the past 10 hours (7).

PRINCIPLE OF THE TEST

The Cocaine Metabolite 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 µl. aliquot of a diluted unknown specimen is incubated with a 100 µl. dilution of enzyme (Horseradish peroxidase) labeled Benzoylecgonine 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 Cocaine Metabolite Direct ELISA Kit avoids extraction of urine or blood sample for measurement. It employs an Benzoylecgonine 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 macromolecules