PRINCIPLE  Do not freeze. The BUP One Step Buprenorphine Test Cassette (Urine) is an  Do not use beyond the expiration date. immunoassay based on the principle of competitive binding. Drugs which may be present in the urine specimen compete against the SPECIMEN COLLECTION AND PREPARATION Buprenorphine Test Cassette (Urine) drug conjugate for binding sites on the antibody. Urine Assay A rapid, one step test for the qualitative detection of During testing, a urine specimen migrates upward by capillary The urine specimen must be collected in a clean and dry container. action. Buprenorphine, if present in the urine specimen below 10 Buprenorphine in human urine. Urine collected at any time of the day may be used. Urine ng/mL, will not saturate the binding sites of antibody-coated specimens exhibiting visible precipitates should be centrifuged, particles in the test. The antibody-coated particles will then be filtered, or allowed settle to obtain a clear specimen for testing. For in vitro diagnostic use only. captured by immobilized Buprenorphine conjugate and a visible Specimen Storage colored line will show up in the test line region. The colored line Urine specimens may be stored at 2-8°C for up to 48 hours prior to Store at 2-30 °c will not form in the test line region if the Buprenorphine level assay. For prolonged storage, specimens may be frozen and stored exceeds 10 ng/mL because it will saturate all the binding sites of below -20°C. Frozen specimens should be thawed and mixed INTENDED USE anti-Buprenorphine antibodies. before testing. The BUP One Step Buprenorphine Test Cassette (Urine) is a lateral flow A drug-positive urine specimen will not generate a colored line in chromatographic immunoassay for the detection of Buprenorphine in the test line region because of drug competition, while a drug- PROCEDURE human urine at a cut-off concentration of 10 ng/mL. negative urine specimen or a specimen containing a drug Allow the test Cassette, urine specimen, and/or controls to reach This assay provides only a qualitative, preliminary analytical test concentration lower than the cut-off will generate a line in the test room temperature (15-30°C) prior to testing. result. A more specific alternate chemical method must be used in line region. To serve as a procedural control, a colored line will 1. Bring the pouch to room temperature before opening it. order to obtain a confirmed analytical result. Gas always appear in the control line region, indicating that proper Remove the test Cassette from the sealed pouch and use it as chromatography/mass spectrometry (GC/MS) or Liquid volume of specimen has been added and membrane wicking has soon as possible. Chromatography/mass spectrometry (LC/MS) are the preferred occurred. 2. Place the test device on a clean and level surface. Hold the confirmatory methods. Clinical consideration and professional dropper vertically and transfer 3 full drops of urine (approx. judgment should be applied to any drug of abuse test result, MATERIALS 100 l) to the specimen well (S) of the test device, and then particularly when preliminary positive results are used. Materials Provided start the timer. Avoid trapping air bubbles in the specimen well  Test Cassette (contains mouse monoclonal anti- (S). See the illustration below. INTRODUCTION Buprenorphine antibody-coupled particles and 3. Wait for the red line(s) to appear. The result should be read at Buprenorphine is a potent analgesic often used in the treatment of Buprenorphine-protein conjugate. A goat antibody is 5 minutes. It is important that the background is clear before opioid addiction. The drug is sold under the trade names employed in the control line system). the result is read. Do not interpret the result after 10 minutes. Subutex,™ Buprenex,™ Temgesic,™ and Suboxone™ which contain  Package insert. Buprenorphine HCl alone or in combination with Naloxone HCl. Materials Required But Not Provided INTERPRETATION OF RESULTS Therapeutically, Buprenorphine is used as a substitution treatment  Specimen collection container (Please refer to the illustration) for opioid addicts. Substitution treatment is a form of medical care  Timer NEGATIVE:* Two distinct colored lines appear. One colored line offered to opiate addicts (primarily heroin addicts) based on a should be in the control line region (C), and another apparent similar or identical substance to the drug normally used. In PRECAUTIONS colored line should be in the test line region (T). This negative substitution therapy, Buprenorphine is as effective as Methadone  For medical and other professional in vitro diagnostic result indicates that the Buprenorphine concentration is below the but demonstrates a lower level of physical dependence. use only. Do not use after the expiration date. detectable level (10 ng/mL). Concentrations of free Buprenorphine and Norbuprenorphine in  The test should remain in the sealed pouch until use. NOTE: urine may be less than 1 ng/mL after therapeutic administration,  All specimens should be considered potentially The shade of color in the test line region (T) will vary, but it should but can range up to 20 ng/mL in abuse situations. The plasma half- hazardous and handled in the same manner as an be considered negative whenever there is even a faint colored life of Buprenorphine is 2-4 hours. While complete elimination of a infectious agent. line. single-dose of the drug can take as long as 6 days, the detection  The used test should be discarded according to local POSITIVE: One colored line appears in the control region (C). window for the parent drug in urine is thought to be regulations. No line appears in the test line region (T). This positive result approximately 3 days. indicates that the Buprenorphine concentration exceeds the The BUP One Step Buprenorphine Test Cassette (Urine) is a rapid STORAGE AND STABILITY detectable level (10 ng/mL). urine screening test that can be performed without the use of an  Store as packaged in the sealed pouch either at room INVALID: Control line (C) fails to appear. Insufficient specimen instrument. The test utilizes a monoclonal antibody to selectively temperature or refrigerated (2-30°C). volume or incorrect procedural techniques are the most likely detect elevated levels of Buprenorphine in urine. The BUP One  The test is stable through the expiration date printed on reasons for control line failure. Review the procedure and repeat Step Buprenorphine Test Cassette (Urine) yields a positive result the sealed pouch. the test using a new test. If the problem persists, discontinue using when Buprenorphine in urine exceeds 10 ng/mL.  The test must remain in the sealed pouch until use. the lot immediately and contact your local distributor. (Urine), the specimens were tested and compared to the self- Buprenorphine above and below the 10 ng/mL cutoff were reported use of Buprenorphine. All specimens, including the ones provided to each site. The following results were tabulated: showing negative results, were then confirmed by LC/MS. The Buprenorphine n Site A Site B Site C following results were tabulated: Concentration per - + - + - + Method Patient Self-Report Total (ng/mL) Site BUP One Results Positive Negative Results 0 15 15 0 15 0 15 0 Step Positive 51 0 51 5 15 15 0 15 0 15 0 Test 7.5 15 8 7 10 5 9 6 Negative 7 150 157 Cassette 12.5 15 0 15 1 14 0 15 Total Results 58 150 208 15 15 0 15 0 15 0 15 % Agreement 88% >99% 97% Effect of Urinary Specific Gravity When compared at 10 ng/mL with LC/MS, the following results Fifteen urine samples with specific gravities ranging from 1.004 to QUALITY CONTROL were tabulated: 1.034 were spiked with Buprenorphine to the concentrations of 5 A procedural control is included in the test. A colored line Method LC/MS Total ng/mL, and 15 ng/mL. The BUP One Step Buprenorphine Test appearing in the control region (C) is considered an internal BUP One Results Positive Negative Results Cassette (Urine) was tested in duplicate using the fifteen neat and procedural control. It confirms sufficient specimen volume, Step Test Positive 55 2 57 spiked urine specimens. The results demonstrate that varying adequate membrane wicking and correct procedural technique. Cassette Negative 1 168 169 ranges of urinary specific gravity do not affect the test results. Control standards are not supplied with this kit; however it is Total Results 56 170 226 Effect of the Urinary pH recommended that positive and negative controls be tested as % Agreement 98% 99% 99% The pH of an aliquoted negative urine pool was adjusted to a pH good laboratory practice to confirm the test procedure and to Analytical Sensitivity range of 5 to 9 in 1 pH unit increments and spiked with verify proper test performance. A drug-free urine pool was spiked with Buprenorphine at the Buprenorphine to 5 ng/mL and 15 ng/mL. The spiked, pH-adjusted following concentrations: 0 ng/mL, 5 ng/mL, 7.5 ng/mL, 10 ng/mL, urine was tested with the BUP One Step Buprenorphine Test LIMITATION 12.5 ng/mL and 15 ng/mL. The result demonstrates >99% accuracy Cassette (Urine) in duplicate. The results demonstrate that varying 1. The BUP One Step Buprenorphine Test Cassette (Urine) at 50% above and 50% below the cut-off concentration. The data ranges of pH do not interfere with the performance of the test. provides only a qualitative, preliminary analytical result. A are summarized below: Cross-Reactivity secondary analytical method must be used to obtain a A study was conducted to determine the cross-reactivity of the confirmed result. Gas chromatography/mass spectrometry Buprenorphine Percent Visual Result Concentration of n test with compounds in either drug-free urine or Buprenorphine (GC/MS) or liquid chromatography/mass spectrometry Negative Positive positive urine. The following compounds show no cross-reactivity 2,3 (ng/mL) Cut-off (LC/MS) are the preferred confirmatory methods. when tested with the BUP One Step Buprenorphine Test Cassette 2. It is possible that technical or procedural errors, as well as 0 0% 90 90 0 5 -50% 90 90 0 (Urine) at a concentration of 100 µg/mL. other interfering substances in the urine specimen may Non Cross-Reacting Compounds 7.5 -25% 90 75 15 cause erroneous results. 4-Acetamidophenol Fenfluramine Oxolinic acid 10 Cut-off 90 60 30 3. Adulterants, such as bleach and/or alum, in urine specimens Acetone Fenoprofen Oxycodone 12.5 +25% 90 31 59 may produce erroneous results regardless of the analytical Acetophenetidin Fentanyl Oxymetazoline 15 +50% 90 0 90 method used. If adulteration is suspected, the test should be Acetylsalicylic acid Fluoxetine Oxymorphone Analytical Specificity repeated with another urine specimen. N- The following table lists compounds that are positively detected in Furosemide Papaverine 4. A positive result indicates presence of the drug or its Acetylprocainamide urine by the BUP One Step Buprenorphine Test Cassette (Urine) at metabolites but does not indicate level of intoxication, Albumin Gentisic acid Pemoline 5 minutes. administration route or concentration in urine. Aminopyrine d-Glucose Penicillin-G 5. A negative result may not necessarily indicate drug-free Conc. Conc. Guaiacol Glyceryl Compound Compound Amitryptyline Pentazocine urine. Negative results can be obtained when drug is present (ng/mL) (ng/mL) Ether but below the cut-off level of the test. Buprenorphine 3-D- Guaiacol Glyceryl Buprenorphine 10 15 Amobarbital Pentobarbital 6. Test does not distinguish between drugs of abuse and certain Glucuronide Ether carbamate medications. Norbuprenorphine 3-D- Norbuprenorphine 20 200 Amoxapine Hemoglobin Perphenazine Glucuronide Amoxicillin Hydralazine Phencyclidine PERFORMANCE CHARACTERISTICS Precision L-Amphetamine Hydrochlorothiazide Phenelzine Accuracy A study was conducted at 3 physician’s offices by untrained Ampicillin Hydrocodone Pheniramine A correlation study was conducted on fifty-eight (58) clinical operators using 3 different lots of product to demonstrate the Apomorphine Hydrocortisone Phenobarbital specimens from patients reporting Buprenorphine use and one- within run, between run and between operator precision. An Aspartame Hydromorphone Phenothiazine hundred fifty (150) urine specimens collected from presumed non- identical panel of coded specimens containing no Buprenorphine, Atropine p- Phentermine drug users. Using the BUP One Step Buprenorphine Test Cassette 25% Buprenorphine above and below the cutoff and 50% Hydroxyamphetami amphetamine Atlas Medical ne (MDA) William James House, Cowley Rd, O-Hydroxyhippuric Trans-2- phenyl D-3,4 Cambridge, CB4 0WX Benzilic acid acid cyclopropylamine Diclofenac Methylendioxy- Thiamine United Kingdom p- methamphetamine Tel: +44 (0) 1223 858 910 Benzoic acid Hydroxymethamphe L-Phenylephrine Thioridazine Fax: +44 (0) 1223 858 524 Dicyclomine Methylphenidate tamine (chlorpromazine) PPI614A01 p- Diflunisal Methyprylon L-Thyroxine Rev C (05.11.2015) Benzoylecgonine Hydroxynorephedrin B-Phenylethylamine Digoxin Methaqualone Tolbutamide Product Reference For in-vitro diagnostic e 4- No. use. Phenylpropanolamin Dimethylaminoantip Metoprolol Cis-Tramadol Benzphetamine Hydroxyzine e (dL-norephedrine) yrine Caution. Store at Bilirubin 3-Hydroxytyramine Noscapine Diphenhydramine Morphine sulfate Trazodone Read product insert Number of tests in dl-Brompheniramine Ibuprofen D,L-Octopamine 5,5- Morphine- Triamterene before use. the pack. Ethyl-p- Diphenylhydantoin Buspirone Orphenadrine aminobenzoate Disopyramide 3-β-D-glucuronide Trifluoperazine Lot (batch) number. Manufacturer. dl- Doxylamine Nalidixic acid Trimethobenzamide Manufacturer Expiry date. Caffeine Etodolac Phenylpropanolamin Ecgonine telephone number. Nalorphine Trimethoprim e hydrochloride Manufacturer fax Cannabidiol Famprofazone Prednisolone Ecgonine methyl number. Naloxone Trimipramine Cannabinol Imipramine Prednisone ester Chloral hydrate Iproniazid 5 beta- EDDP Naltrexone Tryptamine pregnane3alpha17al Efavirenz (Sustiva) Methyprylon D, L-Tryptophan Chloramphenicol l-Isoproterenol pha-21triol 21 EMDP Metoprolol Tyramine Chlordiazepoxide Isoxsuprine Procaine Ephedrine Nimesulide D, L-Tyrosine Chloroquine Kanamycin Promazine (1r,2s)-(-)Ephedrine Norcodeine Uric acid Chlorothiazide Ketamine Promethazine l-ψ-Ephedrine Morphine sulfate Verapamil d-Chlorpheniramine Ketoprofen D,L-Propanolol Alpha- dl-Chlorpheniramine Labetalol D-Propoxyphene dl-Epinephrine Naphthaleneacetic Zomepirac Chlorpromazine L-Ascorbic acid D-Pseudoephedrine Acid Chlorprothixene L-Ephedrine Quinacrine Erythromycin Norethindrone Cholesterol L-Epinephrine Quinidine β-Estradiol Normorphine Cimetidine Levorphanol Quinine Estrone-3-sulfate D-Norpropoxyphene Clomipramine Lidocaine Ranitidine Ethanol (Ethyl Oxazepam Clonidine Lindane Riboflavin alcohol) Cocaine HCl Lithium carbonate Salicylic acid Codeine Loperamide Secobarbital REFERENCES Serotonin (5- 1. Glass, IB. The International Handbook of Addiction Cortisone Maprotiline hydroxytyramine) Behavior. Routledge Publishing, New York, NY. 1991, l-Cotinine Meperidine Sodium chloride 216 Creatinine Mephentermine Sulfamethazine 2. Baselt RC. Disposition of Toxic Drugs and Chemicals in Cyclobarbital Meprobamate Sulindac Man. 6th Ed. Biomedical Publ., Davis, CA.,129, 2002 Cyclobenzaprine Methadone Temazepam 3. Hawks RL, CN Chiang. Urine Testing for Drugs of Abuse. D- Deoxycorticosterone Tetracycline National Institute for Drug Abuse (NIDA), Research Methamphetamine Monograph 73, 1986. L- Tetrahydrocortisone l-Deoxyephedrine Methamphetamine ,3-acetate L-Depreny Methaqualone Tetrahydrozoline Dextromethorphan Methoxyphenamine Thebaine L-3,4- Diazepam Theophylline Methylenedioxy-