Amphetamine 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

Amphetamine ELISA MSDS

Amphetamine ELISA Package Insert

INTENDED USE

The Calbiotech Amphetamine Direct ELISA Kit is a specific and sensitive in-vitro test to detect the presence of d-amphetamine in samples such as whole blood, oral fluids, serum, plasma and urine.

SUMMARY AND EXPLANATION

While the assay will detect amphetamine use, interference by l-amphetamine and pseudo-ephedrine is virtually nonexistent. Amphetamine is a potent central nervous system stimulant. The (+)-isomer also referred to as d-amphetamine is three to four times more potent than the (-)-isomer, l-amphetamine (2). Amphetamine may be metabolized and excreted as the p-hydroxy isomer. Amphetamines act by inducing euphoria, irritability, anxiety and paranoia. Urinary excretion rates are influenced by the urinary pH with acidic urine favoring the excretion of unchanged drug(2). Up to 80% of a given dose may be excreted unchanged, especially in acid urine. Alkaline urine reduces the excretion of unchanged amphetamine to less than 5% of the dose.

PRINCIPLES OF THE TEST

The Amphetamine Direct ELISA Kit (for d-amphetamine measurement) 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 d-amphetamine 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 Amphetamine Direct ELISA Kit avoids extraction of urine sample for measurement. It employs a d-amphetamine 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.