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EDDP (Methadone Metabolite) assay

Product Method Size Catalog Price Quantity
EDDP (Methadone Metabolite) assay Homogenous EIA R1 2 x 16.9ml, R2 2 x 8ml DA4013 $1491.72
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  • Format
    Liquid ready to use
  • Assay Range
  • Working Stability 15-25 °C
  • Working Stability 2-8 °C
    28 days
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Intended Use

For the qualitative and semi-quantitative determination of Methadone Metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in human urine. The cut off for the qualitative application is 300ng/ml. This product is suitable for use on the RX Series instruments, which includes the Rx Daytona and the Rx Imola.

This assay provides only a preliminary result. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly in evaluating a preliminary positive result. To obtain a confirmed analytical result, a more specific alternate chemical method is needed. Gas chromatography/mass spectroscopy (GC/MS) is the recommended confirmatory method.

Clinical Significance

Methadone is a synthetic diphenylheptanonylamine opioid that has a similar analgesic activity and potency to morphine when administered parenterally. However unlike morphine, it reliably retains its effectiveness when given orally, and tolerance and physical dependency develop slowly.

Although methadone is prescribed to relieve chronic pain, its primary medical application is the detoxification and/or maintenance treatment of narcotic or heroin addiction. The abuse potential of methadone is comparable to that of morphine due to its resembling pharmacological activity. Methadone is readily absorbed from gastrointestinal tract when ingested, and metabolized extensively in the liver. Initial N-demethylation results in normethadone metabolite, which rapidly undergoes cyclsation followed by dehydration to form the 2-ethylidene-1,5-dimethyl-3,3- diphenylpyrrolidine, commonly known as EDDP. Further N-demethylation yields a secondary metabolite, the 2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline (EMDP). The metabolites are secreted in urine or bile along with unchanged drug. Current methadone immunoassays can detect only the parent drug and are subject to “false positives” from adulteration of samples for drug of abuse testing, or “false negatives” for methadone compliance testing urine samples from fast metabolizers. As a result, confirmation of the presence of EDDP by LC or GC is often required. EDDP immunoassay detection will make compliance testing easy, and also rule out the possibility adulteration of unsupervised sample collections.


The assay is based on competition between drug in the sample and drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. Enzyme activity decreases upon binding to the antibody, and the drug concentration in the sample is measured in terms of enzyme activity.

In the absence of drug in the sample, methadone metabolite-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. However, when free drug is present in the sample, antibody would bind to free drug; the unbound methadone metabolite-labeled G6PDH then exhibits its maximal enzyme activity.

Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at 340 nm.