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3. Handle all specimens as potentially infectious.
distinguish between drug of abuse and certain foods and medicines.
4. Humidity sensitive product, do not open foil pouch until it is ready to be
The K2/Spice Test test is a qualitative assay. It identifies JWH-018
5. Use a new urine specimen cup for each sample to avoid cross
pentanoic acid or JWH-073 butanoic acid in human urine at a
concentration of 25 ng/mL or higher and other K2 compounds at the
FOR THE QUALITATIVE ASSESSMENT OF SYNTHETIC CANNABIS (K2) IN
6. The immunoassy reaction between anitbody and antigen requires ions
indicated levels. The exact concentration of the synthetic cannabis cannot
to react as designed. Tap water ( or drinkng water) generally speaking does
be determined by this assay. The test is intended to distinguish a negative
result from a presumptive positive result. All positive results must be
not contain these ions and will not create the necessar reaction. Therefore,
water is not a good specimen to use as a negative control test and will
confirmed using an alternate method, preferably GC/MS. NOTE: This
The DrugCheck K2/Spice Test is an immunochromatography-based one
cause the strip to give an erroneous result.
test has been determined to cross react with the drug Lamictal
step in vitro test. It is designed for qualitative determination of synthetic
(Lamotrigine). Specimen from individuals taking this prescription
cannabis JWH-018 and JWH-073’s major metabolites in human urine
drug will likely register a false positive for K2. All presumptive
specimens at cut-off level of 25 ng/ml. In addition, 14 other synthetic
It is required that approximately 120-150 ml of sample for each test.
positive screens should be sent to a confirmation lab for analysis.
cannabis compound metabolities can be detected; compounds and levels
Fresh urine specimens do not need any special handling or treatment.
described in “Specificity”
section of the insert.
Specimens should be collected in a clean, dry, plastic or glass container.
This assay provides only a preliminary analytical test result. A more
If the assay is not performed immediately, urine specimen may be
specific alternative chemical method must be used in order to obtain
refrigerated at 2-8ºC or frozen up to 7 days. Specimens should be
The cut-off concentration (sensitivity level) of K2/Spice Test is determined
a confirmed analytical result. Gas chromatography/mass spectrometry
thawed and brought to room temperature before testing. Urine specimens
to be 25 ng/mL of JWH-018 5-pentanoic acid metabolite and 25 ng/mL
(GC/MS) has been established as the preferred confirmatory method by
exhibiting a large amount of precipitate or turbidity should be centrifuged or
of JWH-073 4-butanoic acid metabolite respectively. Other K2 compounds
the Substance Abuse Mental Health Services Administration (SAMHSA).
allowed to settle before testing. Avoid contact with skin by wearing gloves
can be detected at the concentration level as indicated in the Specificity
Clinical consideration and professional judgment should be applied to any
drug of abuse test result, particularly when preliminary positive results are
Each listed substance that may be found in the urine was evaluated and indicated
negative result at concentration of 100 μg/ml or higher unless it is specified.
Good Laboratory Practice recommends the daily use of control materials
1. The accuracy of the K2/Spice Test was evaluated in K2 spiked urine
to validate the reliability of device. Control materials should be assayed as
specimens. Forty (40) K2 urine specimens were spiked with JWH-018
Synthetic cannabis is a psychoactive herbal and chemical product that,
clinical specimen and challenging to the assay cutoff concentration, e.g.,
pentanoic acid or JWH-73 butanoic acid from 10 to 150 ng/mL; 30
when consumed, mimics the effects of cannabis. It is best known by the
25% above and below cutoff concentration. If control values do not fall
samples with JWH-018 pentanoic acid or JWH-073 butanoic acid
brand name K2 and Spice, both of which have largely become genericized
within established range, assay results are invalid. Control materials which
concentration between 25 and 150 ng/mL were all found positive (100%
trademarks used for refer to any systhetic cannabis product. The studies
are not provided with this test kit are commercially available.
agreement ); 10 samples with JWH-018 pentanoic acid or JWH-073
suggest that synthetic cannabinoid intoxication is associated with acute
The K2/Spice Test provides a built-in process control with a different
butanoic acid concentration between 10 and 18.75 ng/mL were found
psychosis, worsening of previously stable psychotic disorders, and also
antigen/antibody reaction at the control region (C). This control line should
may have the ability to trigger a chronic (long-term) psychotic disorder
always appear regardless the presence of drug or metabolite. If the control
2. A second study was conducted by a third-party laboratory using known
among vulnerable individuals such as those with a family history of mental
line does not appear, the test device should be discarded and the obtained
positive urine from six (6) donors. Each specimen was screened with the
illness. A large and complex variety of synthetic cannabinoids, most ofter
result is invalid. The presence of this control band in the control region
K2/Spice Test. The laboratory then processed and tested each specimen
cannabicyclohexanol, JWH-018, JWH-073, or HU-210, are used. As of
serve as 1) verification that sufficient volume is added, 2) that proper flow
March 1, 2011, five cannabinoids, JWH-018, JWH-073, CP-47, JWH-200
and cannabicyclohexanol are illegal in US because these substances have
the potential to be extremely harmful and, therefore, pose an imminent
1. Bring all materials and specimens to room temperature.
The K2/Spice Test is based on the principle of specific immunochemical
2. Remove the single test strip or device from the sealed foil pouch.
reaction between antibodies and antigens to analyze particular compounds
3. Dip the strip into the urine specimen with the arrow pointing toward
in human urine specimen. The assay relies on the competition for binding
the sample. The sample level should not be higher than the arrow pointed
antibody between drug conjugate and free drug which may be present
in the urine specimen being tested. When drug is present in the urine
specimen, it competes with drug conjugate for the limited amount of
4. Hold the strip in the urine until a reddish color appears at the test area
antibody-dye conjugate. When the amount of drug is equal or more than
The precision study was performed by three individuals observing the test
the cut-off it will prevent the binding of drug conjugate to the antibody.
5. Withdraw the strip and place it face up on a clean, non-absorptive
result to determine the random error of visual interpretation. The test results
Therefore, a positive urine specimen will not show a colored band on the
surface or leave the strip in urine if the urine level is not higher than arrow
were found to have no significant differences between the three observers.
test line zone, indicating a positive result, while the presence of a colored
6. Read the results at 5-10 minutes 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 the control line zone if the test
Test results may not be accurate after 10 minutes.
device is stored in good condition and the test is performed appropriately.
Negative: 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 concentrations of metabolites
2. K2/Spice Test: The amount of each coated antigen and/or antibody on
of JWH-18, JWH-73 or other detection compounds in the specimen is
The specificity for K2/Spice Test test was tested by adding various drugs,
the strip is less than 1.0 mg for antigen conjugate and is less than 1.0 mg
either zero or less than cut-off level.
drug metabolites, and other compounds that are likely to be present in
urine. All compounds were prepared in drug-free normal human urine.
Positive: One colored band forms. One colored band appears in control line
Test zone: contains K2 protein antigen conjugates.
zone. No colored band is found in test line zone. This is an indication that
Control zone: contains goat anti-rabbit IgG antibody.
the concentrations of metabolites of JWH-18, JWH-73 or other detection
The K2/Spice Test performance at cut-off level is not affected when pH
in the specimen is above the cut-off level.
3. Conjugate pad: contains mouse anti-K2 monoclonal antibody.
and Specific Gravity ranges of urine specimen are at 5.0 to 8.0 and 1.005
1. Urine testing for drugs of abuse, NIDA Research Monograph 73 (1986).
Invalid: If there is no colored band in control line zone, the test result is
2. InfoFacts-Club drugs, NIDA , May 2006, http://www.nida.nih.gov/infofacts/
invalid. Retest the sample with a new device.
The following substances were tested and confirmed not to interfere with
Note: A borderline (+/-) in test line zone should be considered negative
K2/Spice Test at the listed concentrations.
3. Drug Fact Sheet, DEA, January 2012, http://www.dea.gov.
4. K2/Spice Test screening and confirmation study, Clinical Reference
The test device should be stored at 4 to 30ºC and will be effective until the
The assay is designed for use with human urine only. A positive result with
expiration date stated on the package. The product is humidity-sensitive
any of the tests indicates only the presence of a drug/metabolite and does
and should be used immediately after being open. Any improperly sealed
There is a possibility that technical or procedural error as well as other
substances in certain foods and medicines may interfere with the test
K2/Spice Test is specific with JWH-018 pentanoic acid and JWH-073
butanoic acid and other compounds described below:
and cause incorrect results. Please refer to the “SPECIFICITY” section for
lists of substances that will produce either positive results, or that do not
2. Do not use the product beyond the expiration date.
interfere with test performance. If a drug/metabolite is found present in
the urine specimen, the assay does not indicate frequency of drug use or
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