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There is a possibility that technical or procedural error as well as other
substances in certain foods and medicines may interfere with the test
and cause incorrect results. Please refer to the “SPECIFICITY” section for
2. Do not use the product beyond the expiration date.
lists of substances that will produce either positive results, or that do not
interfere with test performance. If a drug/metabolite is found present in
FOR THE QUALITATIVE ASSESSMENT OF SYNTHETIC CANNABIS (K2) IN
3. Handle all specimens as potentially infectious.
the urine specimen, the assay does not indicate frequency of drug use or
K2/Spice Test is specific with JWH-018 pentanoic acid and JWH-073
4. Humidity sensitive product, do not open foil pouch until it is ready to be
distinguish between drug of abuse and certain foods and medicines.
The DrugCheck K2/Spice Test is an immunochromatography-based one
5. Use a new urine specimen cup for each sample to avoid cross
The K2/Spice Test test is a qualitative assay. It identifies JWH-018
step in vitro test. It is designed for qualitative determination of synthetic
pentanoic acid or JWH-073 butanoic acid in human urine at a
cannabis JWH-018 and JWH-073’s major metabolites in human urine
6. The immunoassy reaction between anitbody and antigen requires ions
concentration of 25 ng/mL or higher and other K2 compounds at the
specimens at cut-off level of 25 ng/ml. In addition, other synthetic
to react as designed. Tap water ( or drinkng water) generally speaking does
indicated level: MAM2201 (100ng/ml), JWH-398 (200ng/mL), JWH-210
cannabis compound metabolities can be detected, MAM2201 (100ng/ml);
not contain these ions and will not create the necessar reaction. Therefore,
(300ng/mL). The exact concentration of the synthetic cannabis cannot be
JWH-0398 (200ng/mL); and JWH-210 (300ng/mL). This assay has not
water is not a good specimen to use as a negative control test and will
determined by this assay. The test is intended to distinguish a negative
been evaluated in the point of care location and is for forensic use only.
cause the strip to give an erroneous result.
result from a presumptive positive result. All positive results must be
Each listed substance that may be found in the urine was evaluated and
This assay provides only a preliminary analytical test result. A more
confirmed using an alternate method, preferably GC/MS. NOTE: This
indicated negative result at concentration of 100 μg/ml or higher unless
specific alternative chemical method must be used in order to obtain
It is required that approximately 120-150 ml of sample for each test.
test has been determined to cross react with the drug Lamictal
a confirmed analytical result. Gas chromatography/mass spectrometry
Fresh urine specimens do not need any special handling or treatment.
(Lamotrigine). Specimen from individuals taking this prescription
(GC/MS) has been established as the preferred confirmatory method by
Specimens should be collected in a clean, dry, plastic or glass container.
drug will likely register a false positive for K2. All presumptive
the Substance Abuse Mental Health Services Administration (SAMHSA).
If the assay is not performed immediately, urine specimen may be
positive screens should be sent to a confirmation lab for analysis.
Clinical consideration and professional judgment should be applied to any
refrigerated at 2-8ºC or frozen up to 7 days. Specimens should be
drug of abuse test result, particularly when preliminary positive results are
thawed and brought to room temperature before testing. Urine specimens
exhibiting a large amount of precipitate or turbidity should be centrifuged or
allowed to settle before testing. Avoid contact with skin by wearing gloves
The cut-off concentration (sensitivity level) of K2/Spice Test is determined
to be 25 ng/mL of JWH-018 5-pentanoic acid metabolite and 25 ng/mL
Synthetic cannabis is a psychoactive herbal and chemical product that,
of JWH-073 4-butanoic acid metabolite respectively. Other K2 compounds
when consumed, mimics the effects of cannabis. It is best known by the
can be detected at the concentration level as follows: MAM2201(100ng/
brand name K2 and Spice, both of which have largely become genericized
Good Laboratory Practice recommends the daily use of control materials
mL), JWH-398 (200ng/mL) and JWH-210 (300ng/mL).
trademarks used for refer to any systhetic cannabis product. The studies
to validate the reliability of device. Control materials should be assayed as
suggest that synthetic cannabinoid intoxication is associated with acute
clinical specimen and challenging to the assay cutoff concentration, e.g.,
psychosis, worsening of previously stable psychotic disorders, and also
25% above and below cutoff concentration. If control values do not fall
1. The accuracy of the K2/Spice Test was evaluated in K2 spiked urine
may have the ability to trigger a chronic (long-term) psychotic disorder
within established range, assay results are invalid. Control materials which
specimens. Forty (40) K2 urine specimens were spiked with JWH-018
among vulnerable individuals such as those with a family history of mental
are not provided with this test kit are commercially available.
pentanoic acid or JWH-73 butanoic acid from 10 to 150 ng/mL; 30
illness. A large and complex variety of synthetic cannabinoids, most ofter
samples with JWH-018 pentanoic acid or JWH-073 butanoic acid
cannabicyclohexanol, JWH-018, JWH-073, or HU-210, are used. As of
The K2/Spice Test provides a built-in process control with a different
concentration between 25 and 150 ng/mL were all found positive (100%
March 1, 2011, five cannabinoids, JWH-018, JWH-073, CP-47, JWH-200
antigen/antibody reaction at the control region (C). This control line should
agreement ); 10 samples with JWH-018 pentanoic acid or JWH-073
and cannabicyclohexanol are illegal in US because these substances have
always appear regardless the presence of drug or metabolite. If the control
butanoic acid concentration between 10 and 18.75 ng/mL were found
the potential to be extremely harmful and, therefore, pose an imminent
line does not appear, the test device should be discarded and the obtained
result is invalid. The presence of this control band in the control region
serve as 1) verification that sufficient volume is added, 2) that proper flow
2. A second study was conducted by a third-party laboratory using known
positive urine from six (6) donors. Each specimen was screened with the
The K2/Spice Test is based on the principle of specific immunochemical
K2/Spice Test. The laboratory then processed and tested each specimen
reaction between antibodies and antigens to 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
1. Bring all materials and specimens to room temperature.
in the urine specimen being tested. When drug is present in the urine
specimen, it competes with drug conjugate for the limited amount of
2. Remove the single test strip or device from the sealed foil pouch.
antibody-dye conjugate. When the amount of drug is equal or more than
3. Dip the strip into the urine specimen with the arrow pointing toward
the cut-off it will prevent the binding of drug conjugate to the antibody.
the sample. The sample level should not be higher than the arrow pointed
Therefore, a positive urine specimen will not show a colored band on the
test line zone, indicating a positive result, while the presence of a colored
4. Hold the strip in the urine until a reddish color appears at the test area
A control line is present in the test window to work as procedural control.
5. Withdraw the strip and place it face up on a clean, non-absorptive
This colored band should always appear on the control line zone if the test
surface or leave the strip in urine if the urine level is not higher than arrow
device is stored in good condition and the test is performed appropriately.
The precision study was performed by three individuals observing the
test result to determine the random error of visual interpretation. The test
6. Read the results at 5-10 minutes minutes after adding the sample.
results were found to have no significant differences between the three
Test results may not be accurate after 10 minutes.
2. K2/Spice Test: 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
1. Urine testing for drugs of abuse, NIDA Research Monograph 73 (1986).
2. InfoFacts-Club drugs, NIDA , May 2006, http://www.nida.nih.gov/
Test zone: contains K2 protein antigen conjugates.
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
Control zone: contains goat anti-rabbit IgG antibody.
results. The negative result indicates that the concentrations of metabolites
3. Drug Fact Sheet, DEA, January 2012, http://www.dea.gov.
3. Conjugate pad: contains mouse anti-K2 monoclonal antibody.
of JWH-18 and/or JWH-73 in the specimen is either zero or less than
4. K2/Spice Test screening and confirmation study, Clinical Reference
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
The specificity for K2/Spice Test test was tested by adding various drugs,
that the concentrations of metabolites of JWH-18 and/or JWH-73 in the
drug metabolites, and other compounds that are likely to be present in
specimen is above the cut-off level.
urine. All compounds were prepared in drug-free normal human urine.
Invalid: If there is no colored band in control line zone, the test result is
invalid. Retest the sample with a new device.
The K2/Spice Test performance at cut-off level is not affected when pH
Note: A borderline (+/-) in test line zone should be considered negative
and Specific Gravity ranges of urine specimen are at 5.0 to 8.0 and 1.005
The test device should be stored at 4 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
The following substances were tested and confirmed not to interfere with
The assay is designed for use with human urine only. A positive result with
K2/Spice Test at the listed concentrations.
any of the tests indicates only the presence of a drug/metabolite and does
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