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coated membrane and a pad containing the antibody-dye The One Step Cocaine/Benzoyl Ecgonine assay is a rapid, conjugate in a protein matrix containing a 0.1 % sodium azide.
qualitative, competitive binding immunoassay for thedetermination of benzoyl ecgonine in human urine. The test MATERIALS REQUIRED, BUT NOT PROVIDED
provides only preliminary data which should be confirmed by other methods1, 2,3,4 such as gas chromatography / mass spectrophotometry (GC/MS). The preliminary positive resultsshould be justified with compelling clinical consideration and WARNINGS AND PRECAUTIONS
1. For in vitro diagnostic use only.
2. For professional use only.
SUMMARY AND EXPLANATION OF THE TEST
3. Do not use kit beyond expiration date imprinted on the The method employs unique monoclonal antibodies to selectively identify benzoyl ecgonine in test samples with a 4. Urine specimens may be infections. Properly handle and high degree of sensitivity. Cocaine is an alkaloid present in the dispose of all used reaction devices in a biohazard Coca leaves (Erythroxylon coca) and its pharmacological properties, such as stimulating and euphoric effects, have beenknown for many centuries.5 Cocaine has been used medicinally as a local anesthetic agent, but its addictive properties have The reagents contained in the OneStep Cocaine/Benzoyl minimized its modern daily use.6 Cocaine is most often self- Ecgonine test kit should be stored at room temperature (15- administered by IV injection, nasal insufflation, (snorting) and by 30°C) and will be stable until the expiration date.
inhaling vapor from heating of free base (smoking). Theavailability of “crack” (a street form of free base) has increased SAMPLE COLLECTION AND PREPARATION
the use by this latter route.7 Elimination of cocaine is Urine (1.0ml) - the sample must be collected in a clean, dry predominantly controlled by its biotransformation. Very low container, either plastic or glass, without any preservatives.
concentrations of cocaine are detected in urine during the initial Urine specimens may be refrigerated (2 - 8°C) and stored up to several hours, and benzoyl ecgonine, a hydrolytic degradation 48 hours, or frozen (-20°C or colder) prior to assaying. If product, persists in urine at a detectable level for 48 hours.8 samples are refrigerated, they must be equilibrated to room Immunoassay testing has been developed for the determination of benzoyl ecgonine in the urine at the concentration of 300ng/ml set by the National Institute on Drug Abuse.9 ASSAY PROCEDURE
The OneStep Cocaine/Benzoyl Ecgonine test kit is an easy, fast, 1. Bring the test kits to room temperature before opening the and visually read screening method without the need for 2. Remove the reaction device from its foil wrapper by tearingalong the “splice “ and place it on a flat surface.
PRINCIPLE OF THE TEST
3. Fill the urine dropper with sample by holding the dropper The One-Step Cocaine/Benzoyl Ecgonine Test consists of a vertically and dispense two to three (2 – 3) full drops of chromatographic absorbent device in which the drug or drug urine (without air bubbles) into the sample well.
metabolites in the sample compete (with a drug conjugate immobilized on a porous membrane support for limitedantibody sites). As the test sample flows up through the INTERPRETATION OF RESULTS
absorbent device, the labeled antibody-dye conjugate binds tothe free drug in the specimen forming an antibody-antigencomplex. This complex competes with immobilized antigenconjugate in the positive reaction zone and will not produce apink-rose color bend when the drug is higher than the detectionlevel of 300 ng/ml. Unbound dye conjugate binds to thereagent in the negative control zone, producing a pink-rosecolor band, demonstrating that the reagents and device arefunctioning correctly. A negative specimen produces two distinctcolor bands in both the test line and reference line. A positivespecimen produces only one color band in the reference line.
Positive: One pink-rose band appears on the control region.
REAGENTS AND MATERIALS SUPPLIED
No apparent band in the test region (T), in which the benzoyl Each kit contains 20 protected foil pouches. Each pouch ecgonine level is above the detection sensitivity level of 300 contains one “reaction device” and one disposable plastic dropper. The reaction device contains a drug-protein conjugate Negative: In addition to the control band, a pink-rose band
also appears in the test region (T). If this is the case, the REFERENCES
benzoyl ecgonine level is below the detection sensitivity level of 1. Hoyt, D.W., Finnigan, R.E., Nee, T., Shuits, T.F., and Butler, T.J., J. Am. Med. Asso. 258:504 - 509 (1987).
2. MacGregor, R.R., Fowler, J.S., and Wolf, A.P., J. of Invalid: If there is no distinct color band visible both in the
Chromatography, 590:354 - 358 (1992).
“T” and “C” region, or there is a visible band only in the “T” 3. Cone, E.J., Yousefnejad, D., Dickerson, S.L., J. of ForensicSciences, 35 (4): 786 - 791 (1990).
region and not the “C” region, then the test is invalid. It is 4. Cone, E.J., Menchen, S.L., Paul, B.D., Mell, L.D., and Mitchell, recommended that the specimen be retested.
J., J. of Forensic Sciences, 34 (1): 15 - 31 (1989).
NOTE: In order to prevent any incorrect results, the test results
5. Holmstedt, B., and Fredga, A., J. Ethnopharmacol.,3:113 - 147 should not be interpreted after 5 minutes.
6. Inaba, T., Can. J. Physiol. Pharmacol., 67: 1154 - 1157 (1989).
7. Jeffcoat, A.R., Perez-Reyers, M., Hill, J.M., Sadler, B.M., and 1. An internal procedure control has been incorporate into the Cook, C.E., Drug Metabolism and Dispostion, 17 (2) :153 - 159 test to ensure product performance and reliability.
2. Good laboratory practice dictates the use of control 8. Baselt, R.C., and Chang, R., J. Anal. Toxicol., 11:81 –82 (1987).
material to test each product shipment, or whenever 9. Federal Register, 53 (69): 11970-11989 (1988).
necessary, to validate reagent performance and reliabilityCommercial controls are available for the purpose.
1. This product is designed to be used for the detection of
Cocaine and its metabolites in human urine only.
2. Although the OneStep Cocaine/Benzoyl Ecgonine test is
very accurate in detecting the urine benzoyl ecgonine
level, there is a possibility of false results due to the
presence of interfering substance in the urine.
3. The test is a qualitative screening assay and is not
suggested for determining the quantitative benzoyl
ecgonine level of urine, or the level of intoxication.
4. Adulterants, such as bleach or other strong oxidizing
agents, when added to urine specimens, may produce
erroneous test results regardless of the analyst method
used. If adulteration is suspected, obtain another urine
The OneStep Cocaine/Benzoyl Ecgonine test had been
designed for the detection of benzoyl ecgonine in urine at the
detection sensitivity of 300 ng/ml, which is suggested for the
The OneStep Cocaine/Benzoyl Ecgonine test can specifically
detect benzoyl ecgonine in urine. There is no interference by
the following substances at a 10 ug/ml concentration in urine:
Morphine Cannabidiol (+), Amphetamine (+),
Deoxyephedrine, Phencyclidine, and Phenobarbital.
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