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in the test line region because of drug competition, while a drug- should be read at 5 minutes. Do not interpret the result
N.C.S TCA
negative urine specimen or a specimen containing a drug concentration less than the cut-off will generate a line in the test line region. To serve as a procedural control, a colored line will always appear at the control line region indicating that proper volume of specimen has been added and membrane wicking has Tricyclic Antidepressants Test Strip
REAGENTS
The test strip contains mouse monoclonal anti-Tricyclic Antidepressants particles and Tricyclic Antidepressants Package Insert
conjugate coated on the membrane. A goat antibody is employed PRECAUTIONS
A rapid, one step test for the qualitative detection of Tricyclic • For in vitro diagnostic use only. Do not use after the expiration For in vitro diagnostic use only. • The test strip should remain in the sealed pouch until use. INTENDED USE
• All specimens should be considered potentially hazardous and handled in the same manner as an infectious agent. The TCA One Step Tricyclic Antidepressants Test Strip (Urine) • The used test strip should be discarded according to federal, Test Results
is a lateral flow chromatographic immunoassay for the detection of Nortriptyline (metabolites of Tricyclic Antidepressants) in human urine at a cut-off concentration of 1000 ng/mL. STORAGE AND STABILITY
INTERPRETATION OF RESULTS
This assay provides only a qualitative, preliminary analytical
The kit can be stored at room temperature or refrigerated (2-30°C). (Please refer to the illustration above) test result. A more specific alternate chemical method must
The test strip is stable through the expiration date printed on the be used in order to obtain a confirmed analytical result. Gas
sealed pouch. The test strip must remain in the sealed pouch NEGATIVE:* Two lines appear. One red line should be in the
chromatography/mass spectrometry (GC/MS) is the until use. DO NOT FREEZE. Do not use beyond the expiration
control region (C), and another apparent red or pink line should preferred confirmatory method. Clinical consideration and
be in the test region (T). This negative result indicates that the professional judgment should be applied to any drug of
Tricyclic Antidepressants concentration is below the detectable abuse test result, particularly when preliminary positive
SPECIMEN COLLECTION AND PREPARATION
results are used.
* NOTE: The shade of red in the test line region (T) may vary,
Urine Assay
but it should be considered negative whenever there is even a The urine specimen must be collected in a clean and dry TCA (Tricyclic Antidepressants) are commonly used for the container. Urine collected at any time of the day may be used. POSITIVE: One red line appears in the control region (C).
treatment of depressive disorders. TCA overdoses can result in Urine specimens exhibiting visible particles should be No line appears in the test region (T). This positive result profound central nervous system depression, cardiotoxicity and centrifuged, filtered, or allowed to settle to obtain clear specimen indicates that the Tricyclic Antidepressants concentration anticholinergic effects. TCA overdose is the most common cause of exceeds the detectable level (1,000 ng/mL Nortriptyline). death from prescription drugs. TCAs are taken or sometimes by Specimen Storage
INVALID: Control line fails to appear. Insufficient specimen
injection. TCAs are metabolized in the liver, both TCAs and their Urine specimens may be stored at 2-8°C for up to 48 hours prior to volume or incorrect procedural techniques are the most likely metabolites are excreted in urine mostly in the form of metabolites testing. For long-term storage, specimens may be frozen and stored reasons for control line failure. Review the procedure and repeat below -20°C. Frozen specimens should be thawed and mixed before the test using a new test strip. If the problem persists, The TCA One Step Tricyclic Antidepressants Strip (Urine) is a rapid discontinue using the lot immediately and contact your local urine-screening test that can be performed without the use of an instrument. The test utilizes a monoclonal antibody to selectively MATERIALS
detect elevated levels of Nortriptyline in urine. The TCA One Step QUALITY CONTROL
Tricyclic Antidepressants Test Strip (Urine) yields a positive result Materials Provided
A procedural control is included in the test. A red line appearing when the Nortriptyline in urine exceeds 1000 ng/mL. in the control region (C) is considered an internal procedural PRINCIPLE
control. It confirms sufficient specimen volume, adequate Materials Required But Not Provided
membrane wicking and correct procedural technique. The TCA One Step Tricyclic Antidepressants Test Strip (Urine) Control standards are not supplied with this kit; however it is is a rapid chromatographic immunoassay based on the principle recommended that positive and negative controls be tested as of competitive binding. Drugs which may be present in the urine good laboratory testing practice to confirm the test procedure specimen compete against the drug conjugate for binding sites DIRECTIONS FOR USE
Allow the test strip, urine specimen, and/or controls to reach
During testing, a urine specimen migrates upward by capillary LIMITATIONS
room temperature (15-30ºC) prior to testing.
action. Tricyclic Antidepressants, if present in the urine 1. Bring the pouch to room temperature before opening it. 1. The TCA One Step Tricyclic Antidepressants Test Strip specimen below 1000ng/mL, will not saturate the binding sites Remove the test strip from the sealed pouch and use it as soon (Urine) provides only a qualitative, preliminaryanalytical of the antibody coated particles in the test strip. The antibodies result. A secondary analytical method must be used to coated particles will then be captured by immobilized Tricyclic 2. With arrows pointing toward the urine specimen, immerse
obtain a confirmed result. Gas chromatography/mass Antidepressants conjugate and a visible colored line will show the test strip vertically in the urine specimen for at least
spectrometry (GC/MS) is the preferred confirmatory up in the test line region. The colored line will not form in the 10-15 seconds. Do not pass the maximum line (MAX) on
test line region if the Tricyclic Antidepressants level exceeds the test strip when immersing the strip. See the illustration 2. It is possible that technical or procedural errors, as well as 1000ng/mL because it will saturate all the binding sites of anti- other interfering substances in the urine specimen may cause 3. Place the test strip on a non-absorbent flat surface, start the A drug-positive urine specimen will not generate a colored line timer and wait for the red line(s) to appear. The result 3. Adulterants, such as bleach and/or alum, in urine specimens may produce erroneous results regardless of the analytical method used. If adulteration is suspected, the test should be 4. A positive result indicates presence of the drug or its metabolites but does not indicate level of intoxication, administration route or concentration in urine. 5. 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. 6. Test does not distinguish between drugs of abuse and PERFORMANCE CHARACTERISTICS
Precision
A study was conducted at 3 geographically distinct physician’s Accuracy
offices by untrained operators using 3 different lots of product to A side-by-side comparison was conducted using the TCA One demonstrate the within run, between run and between operator Step Tricyclic Antidepressants Test Strip (Urine) and a leading precision. An identical panel of coded specimens containing, commercially available TCA rapid test. Testing was performed according to GC/MS, no Nortriptyline, 25% Nortriptyline above on 300 clinical specimens. Ten percent of the specimens and below the cut-off and 50% Nortriptyline above and below employed were either at –25% or +25% level of the cut-off the 1,000 ng/mL cut-off was provided to each site. For the concentration of Tricyclic Antidepressants. Presumptive positive specimens at -50% cut-off concentration, the 3 sites had a 98% results were confirmed by GC/MS. The following results were agreement. For the –25% to +25% cut-off specimens, the 3 sites demonstrated a 75% agreement. For specimens at +50% cut-off concentration, the 3 sites demonstrated a 93% agreement. The 3 Other TCA Rapid Test
sites demonstrated an overall agreement of 89%. Positive Negative Results
TCA One Step
Effect of Urinary Specific Gravity
Test Strip
Fifteen (15) urine specimens with specific gravity range from 1.001 to 1.032 were spiked with 500 ng/mL and 1,500 ng/mL of Total Results
Nortriptyline respectively. The TCA One Step Tricyclic % Agreement with this
97% 99% 98% Antidepressants Test Strip (Urine) was tested in duplicate using Rapid Test kit
the fifteen neat and spiked urine specimens. The results When compared with GC/MS at a cut-off of 1000ng/ml, the following demonstrate that varying ranges of urinary specific gravity does Method GC/MS
Effect of the Urinary pH
Positive Negative Results
The pH of an aliquoted negative urine pool was adjusted to a pH TCA One Step
143 range of 5 to 9 in 1 pH unit increments and spiked with Test Strip
153 157 Nortriptyline to 500 ng/mL and 1,500 ng/mL. The spiked, pH- BIBLIOGRAPHY
Total Results
adjusted urine was tested with the TCA One Step Tricyclic 1. Rose,J.B.,Tricyclic antidepressants toxicity. J. Toxicity % Agreement with GC/MS
Antidepressants Test Strip (Urine) in duplicate. The results Analysis
demonstrate that varying ranges of pH does not interfere with 2. Urine Testing for Drugs of Abuse, National Institute on Analytical Sensitivity
Drug-Abuse (NIDA), Research Monogragh 73 ,1986. Cross-Reactivity
3. Fed. Register, Department of Health and Human Services, A drug-free urine pool was spiked with Tricyclic A study was conducted to determine the cross-reactivity of the Mandatory Guidelines for Fedaral Workplace Drug Testing Antidepressants at the following concentrations: 0 ng/mL, 500 test with compounds in either drug-free urine or Nortriptyline- ng/mL, 750 ng/mL, 1,000 ng/mL, 1,250 ng/mL and 1,500 positive urine. The following compounds show no cross- ng/mL. The result demonstrates >99% accuracy at 50% above reactivity when tested with the TCA One Step Tricyclic and 50% below the cut-off concentration. The data are Antidepressants Test Strip (Urine) at a concentration of 100 Nortriptyline
Visual Result
Non Cross-Reacting Compounds
Percent of
Concentratio
Acetamidophenol Estrone-3-sulfate Oxolinic n (ng/mL)
Nova Century Scientific, Inc.
22 8 Aminopyrine Furosemide Penicillin-G 5 25 Amoxicillin Hemoglobin Pentobarbital 0 30 Ampicillin Hydralazine Perphenazine Analytical Specificity
The following table lists compounds that are positively detected in urine by the TCA One Step Tricyclic Antidepressants Test Compound Concentration

Source: http://www.novacentury.com/inserts/insert%2022.pdf

Pbio.1000412 1.5

Improving Bioscience Research Reporting: The ARRIVEGuidelines for Reporting Animal ResearchCarol Kilkenny1*, William J. Browne2, Innes C. Cuthill3, Michael Emerson4, Douglas G. Altman51 The National Centre for the Replacement, Refinement and Reduction of Animals in Research, London, United Kingdom, 2 School of Veterinary Science, University ofBristol, Bristol, United Kingdom, 3 School of Biol

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