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Microsoft word - k2 product insert for whpm 75ng ml version 3.0 9th revision.doc
One Step K2 Drug of Abuse Test
For Forensic Use Only
The One Step K2 Drug of Abuse Test is a lateral flow chromatographic immunoassay for the qualitative detection of synthetic cannabinoids metabolites in human urine specimen at the cut-off level of 50ng/mL. This assay is intended for forensic use only. This assay provides only a preliminary qualitative test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Liquid chromatography/mass spectrometry (LC/MS) is the preferred confirmatory method. Apply clinical and professional judgment to any drug of abuse test result, particularly when preliminary positive results are obtained.
SUMMARY AND EXPLANATION OF THE TEST
Synthetic Cannabinoids (K2 50)
Since 2004, herbal mixtures such as ‘Spice’ have been sold in Switzerland, Austria, Germany and other European countries mainly via Internet shops. Although declared as incense, they are smoked as ‘bio-drugs’ by the consumers. In corresponding blogs, drug users reported cannabis-like effects after smoking. These products enjoy great popularity particularly among younger people, as up to now the mixtures are sold in head shops and via internet in many countries without age restriction. 1 JWH-018 was developed and evaluated in basic scientific research to study structure activity relationships related to the cannabinoid receptors. 2 JWH-073 has been identified in numerous herbal products, such as “Spice”, “K2”, and K3”. 3 These products may be smoked for their psychoactive effects. The One Step K2 Drug of Abuse Test yields a positive result when synthetic cannabinoid compounds in urine exceed 50ng/mL.
The One Step K2 Drug of Abuse Test is an immunoassay based on the principle of competitive binding. Drug which may be present in the urine specimen compete against their respective drug conjugate for binding sites on their specific antibody. During testing, a urine specimen migrates upward by capillary action. A drug, if present in the urine specimen below its cut-off concentration, wil not saturate the binding sites of its specific antibody. The antibody wil then react with the drug-protein conjugate and a visible colored line wil show up in the test line region of the specific drug strip. The presence of drug above the cut-off concentration wil saturate al the binding sites of the antibody. Therefore, the colored line wil not form in the test line region. A drug-positive urine specimen will not generate a colored line in the specific test line region of the strip because of drug competition, while a drug-negative urine specimen will generate a line in the test line region because of the absence of drug competition. To serve as a procedural control, a colored line wil always appear in the control line region, indicating that proper volume of specimen has been applied and membrane wicking has occurred.
The test contains a membrane strip coated with drug-protein conjugate (purified bovine albumin) at the test line, a goat polyclonal antibody against gold-protein conjugate at the control line, and a dye pad which contains colloidal gold particles coated with mouse monoclonal K2 antibody.
• For Forensic Use Only. • Do not use after the expiration date. • The test panel should remain in the sealed pouch until use. • Use of gloves is recommended to avoid unnecessary contact with the specimen. • The used test device and urine specimen should be discarded according to federal, state and local regulations. • The test is for single use.
STORAGE AND STABILITY
Store as packaged in the sealed pouch at 2⁰C - 30⁰C (36⁰F - 86⁰F). The test is stable through the expiration date printed on the sealed pouch. The test device must remain in the sealed pouch until use. DO NOT FREEZE.
SPECIMEN COLLECTION AND PREPARATION
The urine specimen must be collected in a clean and dry container. Urine collected at any time of the day may be used. Urine specimens exhibiting visible precipitates should be allowed to settle to obtain a clear specimen for testing.
Urine specimen collected for later testing may be stored at 2⁰C - 8⁰C (36⁰F - 46⁰F) for up to 48 hours. For prolonged storage, specimens may be frozen and stored below -20⁰C. Frozen specimens should be thawed and mixed well before testing.
• Test device
• Package insert Materials Required But Not Provided:
• Specimen collection container • Disposable gloves • Timer
INSTRUCTIONS FOR USE]
1) Remove the test device from the foil pouch. 2) Remove the cap from the test device. Label the device with patient or control identifications. 3) Immerse the absorbent tip into the urine sample for 5 seconds. Urine sample should not touch the plastic device. 4) Replace the cap over the absorbent tip and lay the device flatly on a non-absorptive clean surface. 5) Read results at 5 minutes
DO NOT INTERPRET RESULT AFTER 10 MINUTES.
INTERPRETATION OF RESULTS
(Please refer to the previous illustration) NEGATIVE:
Two lines appear. * One color line should be in the control region (C), and another apparent color line adjacent should be in the test region (T). This negative result indicates
that the drug concentration is below the detectable level.
*NOTE: The shade of color in the test line region (T) will vary, but it should be considered negative whenever there is even a faint distinguishable color line. POSITIVE:
One color line appears in the control region (C). No line appears in the test region (T). This positive result indicates that the drug concentration is above the detectable level. INVALID:
Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and
repeat the test using a new test device. If the problem persists, discontinue using the lot immediately and contact your supplier.
A procedural control is included in the test. A color line appearing in the control region (C) is considered an internal procedural control. It confirms sufficient specimen volume, adequate membrane wicking and correct procedural technique.
1. The One Step K2 Drug of Abuse Test provides only a qualitative, preliminary analytical result. A secondary analytical method must be used to obtain a confirmed result.
Liquid chromatography/mass spectrometry (LC/MS) is the preferred confirmatory method.
2. There is a possibility that technical or procedural errors, as well as other interfering substances in the urine specimen may cause erroneous results. 3. A positive result does not indicate intoxication of the donor, the concentration of drug in the urine, or the route of drug administration. 4. A negative result may not necessarily indicate drug-free urine. Negative results can be obtained when drug is present but below the cut-off level of the test. 5. If adulteration is suspected, the test should be repeated with another new urine specimen and a new test device.
The accuracy of the One Step K2 Drug of Abuse Test was evaluated in comparison to liquid chromatography/mass spectrometry (LC/MS) or gas chromatography/mass spectrometry (GC/MS). 60 specimens, comprised of 20 negative urine samples and 40 positive urine samples, were blinded and tested with the One Step K2 Drug of Abuse Test and compared to LC/MS results. The testing showed a >95% agreement between two methods.
A study was conducted in an effort to determine the precision of the One Step K2 Drug of Abuse Test. Testing was conducted using three different lots of product to demonstrate the within-run and between-run precision. The correlation with expected results for the solutions targeted to +/-50% of the cut-off was > 99% across al lots.
The cut-off concentration (sensitivity level) of K2/Spice test is determined to be 50ng/mL of JWH-018 Pentanoic Acid Metabolite and 50ng/mL of JWH-073 Butanoic Acid Metabolite respectively. Testing should be run in 10 replicates with negative urine and standard control at ±25% cut-off and ±50% cut-off concentration levels. Test results are summarized below.
The fol owing table lists the concentration of compounds (ng/mL) that were detected positive in urine by the One Step K2 Drug of Abuse Test at a reading time of 5 to 10 minutes.
EFFECT OF SPECIMEN SPECIFIC GRAVITY
The urine samples of normal, high, and low specific gravity ranges from 1.000-1.025 were spiked with drugs at 50% below and 50% above cut-off levels respectively and tested using One Step K2 Drug of Abuse Test. The results demonstrate that varying ranges of specimen specific gravity do not interfere with the performance of the test
EFFECT OF SPECIMEN PH
The pH of an aliquot negative urine pool was adjusted to pH ranges of 4.5 - 9.0, and spiked with drugs at 50% below and 50% above cut-off levels. The spiked, pH-adjusted urine was tested with the One Step K2 Drug of Abuse Test. The results demonstrate that varying ranges of specimen pH do not interfere with the performance of the test.
A study was conducted to determine the cross-reactivity of the test with compounds in either drug-free urine or drug positive urine containing K2. The fol owing compounds show no
cross-reactivity when tested with the One Step K2 Drug of Abuse Test at concentrations of 100µg/mL. K2 NON CROSS-REACTING COMPOUNDS
1. Auwarter V et. al. ‘Spice’ and other herbal blends: harmless incense or cannabinoid designer drugs? J. Mass Spectrom.
44: 832-837 (2009). 2. U.S Drug Enforcement Administration (DEA). Drugs and Chemicals of Concern: JWH-073. (2009). 3. U.S. Drug Enforcement Administration (DEA). Drugs and Chemicals of Concern: JWH-018. (2009). Manufactured by: W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706
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