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Microsoft word - r13-470 tramadol tc 12-08.doc


MATERIALS PROVIDED
2. Tramadol test device. The amount of each coated antigen and/or antibody on the strip is less than 1.0 mg for antigen conjugate and is less than 1.0 mg for goat anti-mouse IgG Test zone: contains tramadol bovine protein antigen Control zone: contains Goat anti-mouse IgG antibody. 3. Conjugate pad: contains mice monoclonal anti-drug MATERIALS REQUIRED BUT NOT PROVIDED
TRAMADOL TEST CASSETTE
FOR THE QUALITATIVE ASSESSMENT OF TRAMADOL STORAGE AND STABILITY
The test device should be stored at 2 to 30ºC and will be effective until the expiration date stated on the package. The product is humidity-sensitive and should be used immediately after being open. Any improperly sealed product should be PRECAUTIONS
1. For in vitro diagnostic and forensic use only. 2. Do not use the product beyond the expiration date. INTENDED USE
3. Handle all specimens as potentially infectious. 4. Humidity sensitive product, do not open foil pouch until it is immunochromatography based one step in vitro test. It is designed for qualitative determination of the tramadol and its 5. Use a new urine specimen cup for each sample to avoid metabolites in human urine specimens at cut-off level of 200 ng/ml. This assay has not been evaluated in the point of care SPECIMEN COLLECTION AND PREPARATION
location and is for use by Healthcare Professionals only. It is required that approximately 150 µl of sample for each test. Fresh urine specimens do not need any special handling or This assay provides only a preliminary analytical test result. A treatment. Specimens should be collected in a clean, dry, plastic more specific alternative chemical method must be used in order or glass container. If the assay is not performed immediately, to obtain a confirmed analytical result. Gas chromatography/ urine specimen may be refrigerated at 2-8ºC or frozen up to 7 mass spectrometry (GC/MS) has been established as the days. Specimens should be thawed and brought to room preferred confirmatory method by the Substance Abuse Mental temperature before testing. Urine specimens exhibiting a large Health Services Administration (SAMHSA). Clinical consideration amount of precipitate or turbidity should be centrifuged or and professional judgment should be applied to any drug of allowed to settle before testing. Avoid contact with skin by abuse test result, particularly when preliminary positive results wearing gloves and proper laboratory attire. QUALITY CONTROL
SUMMARY AND EXPLANATION
Good Laboratory Practice recommends the daily use of control Tramadol is a quasi-narcotic analgesic used in the treatment of materials to validate the reliability of device. Control materials moderate to severe pain. It is a synthetic analog of codeine, but should be assayed as clinical specimen and challenging to the has a low binding affinity to the mu-opioid receptors. Large assay cutoff concentration, e.g., 25% above and below cutoff doses of tramadol can develop tolerance and physiological concentration. If control values do not fall within established dependency and lead to its abuse. Tramadol is extensively range, assay results are invalid. Control materials which are not metabolized after oral administration. Approximately 30% of the provided with this test kit are commercially available. dose is excreted in the urine as unchanged drug, whereas 60% Tramadol test provides a built-in process control with a different is excreted as metabolites. The major pathways appear to be N- antigen/antibody reaction at the control region (C). This control and O- demethylation, glucoronidation or sulfation in the liver. line should always appear regardless the presence of drug or metabolite. If the control line does not appear, the test device TEST PRINCIPLE
should be discarded and the obtained result is invalid. The Tramadol test cassette is based on the principle of specific presence of this control band in the control region serve as 1) immunochemical reaction between antibodies and antigens to verification that sufficient volume is added, 2) that proper flow analyze particular compounds in human urine specimen. The assay relies on the competition for binding antibody between drug conjugate and free drug which may be present in the urine PROCEDURE
specimen being tested. When drug is present in the urine 1. Bring all materials and specimens to room temperature. specimen, it competes with drug conjugate for the limited 2. Remove the test cassette from the sealed foil pouch. amount of antibody-dye conjugate. When the amount of drug is 3. Place the transfer pipette in the specimen and depress the equal or more than the cut-off, 200ng/ml, it will prevent the binding of drug conjugate to the antibody. Therefore, a positive 4. Hold the pipette in a vertical position over the sample well urine specimen will not show a colored band on the test line of the test cassette and deliver 3 drops (120-150 µl) of zone, indicating a positive result, while the presence of a colored 5. Read the results at 5 minutes after adding the sample. A control line is present in the test window to work as procedural control. This colored band should always appear on INTERPRETATION OF RESULTS
the control line zone if the test device is stored in good condition Negative:
and the test is performed appropriately. Two colored bands form. The appearance of two colored bands, one in test line zone and the other in control line zone, indicates negative results. The negative result indicates that the tramadol ------------------------------------------------------------------------ concentration in the specimen is either zero or less than cut-off -------------------------------------------------- Positive:
------------------------------------------------------------------------ One colored band forms. One colored band appears in control -------------------------------------------------- line zone. No colored band is found in test line zone. This is an indication that the tramadol level in the specimen is above the Invalid:
------------------------------------------------------------------------ If there is no colored band in control line zone, the test result is --------------------------------------------------- invalid. Retest the sample with a new device. Each listed substance that commonly found in the urine was evaluated and indicated negative result at concentration of 100 Note: A borderline (+/-) in test line zone should be considered LIMITATION OF PROCEDURE
The assay is designed for use with human urine only. A positive result with any of the tests indicates only the presence of a drug/metabolite and does not indicate or measure intoxication. There is a possibility that technical or procedural error as well other substances in certain foods and medicines may interfere with the test and cause false results. Please refer “SPECIFICITY” section for lists of substances that will produce either positive results, or that do not interfere with test Phencyclidine Phenylethylamine-α Phenylpropanolamine performance. If a drug/metabolite is found present in the urine 11-nor-∆8 –THC-9-COOH 11-nor-∆8 –THC-9-COOH specimen, the assay does not indicate frequency of drug use or distinguish between drug of abuse and certain foods and EXPECTED RESULTS
Immunospec Tramadol Test is a qualitative assay. It identifies REFERENCES
tramadol in human urine at a concentration of 200 ng/ml or 1. Urine testing for drugs of abuse, NIDA Research higher. The concentration of the tramadol cannot be determined by this assay. The test is intended to distinguish negative result 2. Drug Facts and Comparisons, 55th ed. St. Louis: Facts from presumptive positive result. All positive results must be confirmed using an alternate method, preferably GC/MS. 3. Raffa, R. et al. Mechanism of action study , J. of Pharmacol. Exp. Ther. Vol. 267, 331 ( 1993) PERRFOMANCE CHARACTERISTICS
A. Sensitivity
The cut-off concentration (sensitivity level) of tramadol test is European Authorized Representative:
CEpartner4U , Esdoornlaan 13, 3951DB Maarn . The Netherlands. Tel.: +31 (0)6.516.536.26 B. Precision
The precision study was performed by three individuals observing the test result to determine the random error of visual interpretation. The test results were found to have no significant differences between the three observers. Manufacturer:
C. Specificity
The specificity for tramadol test was tested by adding various drugs, drug metabolites, and other compounds that are likely to be present in urine. All compounds were prepared in drug-free 1. Interference testing
Tramadol test performance at cut-off level is not affected when pH and Specific Gravity ranges of urine specimen are at 4.5 to The following substances were tested and confirmed not to interfere with tramadol test at the listed concentrations.
2. Specificity

The following table lists compounds that are detected by tramadol test which produced positive results when tested at levels equal or greater than the concentrations listed below:

Source: http://immunospec.com/files/R13-470-Tramadol-Cassette-12-08.pdf

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