(110 days)
Not Found
No
The device description and performance studies focus on a standard immunoassay technique and do not mention any AI or ML components. The analysis is based on enzyme activity and spectrophotometric measurement, which are traditional laboratory methods.
No.
This device is an in vitro diagnostic device used to determine the presence of tramadol in human urine, not to treat a medical condition.
Yes
The "Intended Use / Indications for Use" section explicitly states, "This in vitro diagnostic device is for prescription use only." This directly indicates its diagnostic purpose.
No
The device is an in vitro diagnostic assay consisting of chemical reagents (R1 and R2) used with automated clinical chemistry analyzers, which are hardware. It is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states: "This in vitro diagnostic device is for prescription use only."
- Device Description: The description details a "homogeneous enzyme immunoassay technique used for the analysis of tramadol in human urine." This describes a test performed in vitro (outside the body) on a biological sample (urine) to diagnose or provide information about a condition (presence of tramadol).
- Sample Type: The device analyzes "human urine," which is a biological specimen.
- Purpose: The assay is intended for the "qualitative and/or semiquantitative determination of tramadol in human urine," which is a diagnostic measurement.
- Care Setting: The intended users are "laboratories with automated clinical chemistry analyzers," which are typical settings for in vitro diagnostic testing.
N/A
Intended Use / Indications for Use
The ARK Tramadol Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of tramadol in human urine at a cutoff concentration of 100 ng/mL. The assay is intended for use in laboratories with automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only.
The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Tramadol Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
Product codes (comma separated list FDA assigned to the subject device)
DJG
Device Description
The ARK Tramadol Assay is a homogeneous enzyme immunoassay technique used for the analysis of tramadol in human urine. The assay is based on competition between drug in the specimen and drug labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. As the latter binds antibody, enzyme activity decreases. In the presence of drug from the specimen, enzyme activity increases and is directly related to the drug concentration. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay.
The ARK Tramadol Assay consists of reagents R1 anti-tramadol rabbit polyclonal antibody with substrate and R2 tramadol derivative labeled with bacterial recombinant G6PDH enzyme.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
human urine
Indicated Patient Age Range
Not Found
Intended User / Care Setting
laboratories with automated clinical chemistry analyzers
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision:
Precision studies were performed using CLSI EP05-A3 as a guideline. Drug-free, negative human urine was supplemented with tramadol (0.0 to 200.0 ng/mL). Each level was assayed in quadruplicate twice a day for 20 days (N=160) in both qualitative and semiquantitative modes.
Qualitative Precision Results:
- 0.0 ng/mL: 160 Negative
- 25.0 ng/mL: 160 Negative
- 50.0 ng/mL: 160 Negative
- 75.0 ng/mL: 160 Negative
- 100.0 ng/mL: 83 Negative/77 Positive
- 125.0 ng/mL: 160 Positive
- 150.0 ng/mL: 160 Positive
- 175.0 ng/mL: 160 Positive
- 200.0 ng/mL: 160 Positive
Semiquantitative Precision Results (Mean ng/mL):
- 0.0 ng/mL: 1.7 (160 Negative)
- 25.0 ng/mL: 29.2 (160 Negative)
- 50.0 ng/mL: 53.7 (160 Negative)
- 75.0 ng/mL: 76.7 (160 Negative)
- 100.0 ng/mL: 98.5 (97 Negative/63 Positive)
- 125.0 ng/mL: 120.5 (160 Positive)
- 150.0 ng/mL: 142.6 (160 Positive)
- 175.0 ng/mL: 165.3 (160 Positive)
- 200.0 ng/mL: 189.0 (160 Positive)
Analytical Recovery:
Recovery across the assay range (50.0 to 1000.0 ng/mL) was assessed using the semiquantitative mode. Drug-free, negative human urine was supplemented with tramadol (1100.0 ng/mL) and dilutions were made proportionally with drug-free human urine. Percentage recovery was calculated based on the mean concentration (N=6) compared to the expected concentration. Recovery ranged from 90.4% to 109.1%.
Analytical Specificity - Tramadol Metabolites:
- O-Desmethyltramadol: 16.67% cross-reactivity at 600 ng/mL
- N-Desmethyltramadol: 66.67% cross-reactivity at 150 ng/mL
Analytical Specificity - Structurally Related Compounds:
Various structurally related compounds were tested and found to be negative at specified concentrations in both qualitative and semiquantitative modes.
Interference – Structurally Unrelated Compounds:
High concentrations of structurally unrelated compounds were added into urine spiked with tramadol (± 25% of the cutoff concentration) and tested in both qualitative and semiquantitative modes. The substances listed did not yield a false result relative to the 100 ng/mL cutoff.
Interference – Endogenous Substances:
High concentrations of endogenous substances were added into urine spiked with tramadol (± 25% of the cutoff concentration). No interference was observed when tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes.
Interference – Specific Gravity and pH:
Urine samples with specific gravity values from 1.000 to 1.030 and pH values ranging from 3.0 to 11.0 were tested in the presence of the two levels of tramadol at ± 25% of the cutoff concentration. No interference was observed.
Interference – Boric Acid:
1% w/v of boric acid was added into urine spiked with tramadol (± 25% of the cutoff concentration). Boric acid interfered with results (Negative in qualitative mode at +25% Cutoff).
Method Comparison:
A total of 115 unaltered clinical human urine specimens were analyzed with the ARK Tramadol Assay in both qualitative and semiquantitative modes and compared to LC-MS/MS.
Results compared to LC-MS/MS:
- Negative results were 50 in Low Negative group ( 150 ng/mL).
- Positive results were 0 in Low Negative group, 5 in Near Cutoff Negative, 4 in Near Cutoff Positive, and 56 in High Positive.
Discordant Results (5 samples) were positive by ARK Immunoassay but negative by LC-MS/MS (ranging from 74.0 to 98.9 ng/mL by LC-MS/MS). O-desmethyltramadol was detected in these samples, contributing to the positive result.
Calibration Curve Stability:
A stored calibration curve was effective up to at least 30 days based on supporting data.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Not Found (Specific metrics such as sensitivity, specificity, PPV, NPV are not explicitly stated, but raw data for method comparison is provided.)
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Immunalysis Tramadol Urine Enzyme Immunoassay K141803
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 862.3650 Opiate test system.
(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (e.g., programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).
0
Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: on the left, there is a symbol representing the Department of Health & Human Services - USA, and on the right, there is the text "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue. The FDA logo is a recognizable symbol associated with the agency responsible for regulating and supervising the safety of food, drugs, and other products in the United States.
December 10, 2018
ARK Diagnostics, Inc. Cherry Mun Manager, Quality and Regulatory Affairs 48089 Fremont Boulevard Fremont, CA 94538
Re: K182280
Trade/Device Name: ARK Tramadol Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: DJG Dated: November 12, 2018 Received: November 13, 2018
Dear Cherry Mun:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's
1
Page 2
requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
for Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known)
Form Approved: OMB No. 0910-0120 Expiration Date: 06/30/2020 See PRA Statement below.
Device Name ARK Tramadol Assay
Indications for Use (Describe)
The ARK Tramadol Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of tramadol in human urine at a cutoff concentration of 100 ng/mL. The assay is intended for use in laboratories with automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only.
The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Tramadol Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| ------------------------------------------------- | -- |
Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
3
510(k) SUMMARY
This 510(k) Summary of Safety and Effectiveness information is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.
The assigned 510(k) number is K182280.
| 807.92 (a)(1): Name: | ARK Diagnostics, Inc. |
|---|---|
| Address: | 48089 Fremont Blvd |
| Fremont, CA 94538 USA | |
| Owner Operator Number: | 10027663 |
| Establishment Registration: | 3005755244 |
| Phone: | (510) 270-6270 |
| FAX: | (510) 270-6298 |
| Contact: | Cherry Mun – (510) 270-6288 |
| Manager of Quality and Regulatory Affairs |
Date Prepared: November 27th, 2018
807.92 (a)(2): Device Name – Trade Name, Common Name, and Classification
| Trade Name: | ARKTM Tramadol Assay |
|---|---|
| Common Name: | Homogeneous Enzyme Immunoassay, Opiate Test System |
Classification:
| Product Code | Classification | Regulation Section | Panel |
|---|---|---|---|
| DJG | Class II | 21 CFR 862.3650 | |
| Opiate Test System | Toxicology | ||
| (91) |
807.92 (a)(3): Identification of the Legally Marketed Predicate Device
Immunalysis Tramadol Urine Enzyme Immunoassay K141803
4
807.92 (a)(4): Device Description
The ARK Tramadol Assay is a homogeneous enzyme immunoassay technique used for the analysis of tramadol in human urine. The assay is based on competition between drug in the specimen and drug labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. As the latter binds antibody, enzyme activity decreases. In the presence of drug from the specimen, enzyme activity increases and is directly related to the drug concentration. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay.
The ARK Tramadol Assay consists of reagents R1 anti-tramadol rabbit polyclonal antibody with substrate and R2 tramadol derivative labeled with bacterial recombinant G6PDH enzyme.
807.92 (a)(5): Intended Use / Indications for Use
ARK Tramadol Assay
The ARK Tramadol Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of tramadol in human urine at a cutoff concentration of 100 ng/mL. The assay is intended for use in laboratories with automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only.
The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Tramadol Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
5
807.92 (a)(6): Technological Similarities and Differences to the Predicate
SUBSTANTIAL EQUIVALENCE COMPARATIVE TABLE
Comparison between the Immunalysis Tramadol Urine Enzyme Immunoassay and the ARK™ Tramadol Assay
| Characteristic | Predicate Device
Immunalysis Tramadol Urine Enzyme
Immunoassay (K141803) | Candidate Device
ARK™ Tramadol Assay |
|-----------------------------------|-----------------------------------------------------------------------------------------------------------------------------|-----------------------------------------|
| Similarities | | |
| Test System | Homogenous enzyme immunoassay
(EIA) | Same |
| Intended Use | For the qualitative and semiquantitative
determination of tramadol in human
urine; For in vitro diagnostic use | Same |
| Sample Matrix | Human urine | Same |
| User Environment | Clinical laboratories; Prescription use
only | Same |
| Mass Spectrometry
Confirmation | Required to confirm preliminary positive
analytical results | Same |
| Platform Required | Automated clinical chemistry analyzer | Same |
| Reagents Form | Liquid - Ready to use | Same |
| Reagent Materials | Two (2) reagent system:
Antibody/substrate reagent and enzyme
labeled conjugate
Sodium azide preservative | Same |
| Storage | 2-8°C until expiration date | Same |
| Measured Analyte | Tramadol | Same |
| Antibody | Polyclonal antibodies to tramadol | Same |
| Detection | Absorbance change measured
spectrophotometrically at 340 nm | Same |
| Characteristic | Predicate Device
Immunalysis Tramadol Urine Enzyme
Immunoassay (K141803) | Candidate Device
ARK™ Tramadol Assay |
| Differences | | |
200 ng/mL
Cutoff Level
100 ng/mL
6
807.92 (b)(1) and 807.92 (b)(2): Brief Description of Nonclinical and Clinical Data
The following performance characteristics were obtained on the Beckman Coulter AU680® automated clinical chemistry analyzer.
Precision
Precision studies were performed using CLSI EP05-A3 as a guideline. Drug-free, negative human urine was supplemented with tramadol (0.0 to 200.0 ng/mL). Each level was assayed in quadruplicate twice a day for 20 days (N=160) in both qualitative and semiquantitative modes. Results are summarized in the tables below.
Qualitative Precision
| Human Urine
(ng/mL) | % Cutoff | # of
Determinations | Qualitative
Precision Results |
|------------------------|----------|------------------------|----------------------------------|
| 0.0 | -100 | 160 | 160 Negative |
| 25.0 | -75 | 160 | 160 Negative |
| 50.0 | -50 | 160 | 160 Negative |
| 75.0 | -25 | 160 | 160 Negative |
| 100.0 | Cutoff | 160 | 83 Negative/
77 Positive |
| 125.0 | +25 | 160 | 160 Positive |
| 150.0 | +50 | 160 | 160 Positive |
| 175.0 | +75 | 160 | 160 Positive |
| 200.0 | +100 | 160 | 160 Positive |
Semiquantitative Precision
| Human
Urine
(ng/mL) | Relative %
Cutoff | # of Results | Mean
(ng/mL) | Semiquantitative
Precision Results |
|---------------------------|----------------------|--------------|-----------------|---------------------------------------|
| 0.0 | -100 | 160 | 1.7 | 160 Negative |
| 25.0 | -75 | 160 | 29.2 | 160 Negative |
| 50.0 | -50 | 160 | 53.7 | 160 Negative |
| 75.0 | -25 | 160 | 76.7 | 160 Negative |
| 100.0 | Cutoff | 160 | 98.5 | 97 Negative/
63 Positive |
| 125.0 | +25 | 160 | 120.5 | 160 Positive |
| 150.0 | +50 | 160 | 142.6 | 160 Positive |
| 175.0 | +75 | 160 | 165.3 | 160 Positive |
| 200.0 | +100 | 160 | 189.0 | 160 Positive |
7
Analytical Recovery
Recovery across the assay range was assessed using the semiquantitative mode. Drug-free, negative human urine was supplemented with tramadol (1100.0 ng/mL) and dilutions were made proportionally with drug-free human urine. Tramadol concentrations ranged from 50.0 to 1000.0 ng/mL. At each level, percentage recovery was calculated based on the mean concentration (N=6) compared to the expected concentration. Results are summarized in the table below.
| Theoretical
Concentration
(ng/mL) | Mean
Concentration
(ng/mL) | Recovery
(%) |
|-----------------------------------------|----------------------------------|-----------------|
| 50.0 | 52.8 | 105.6 |
| 100.0 | 107.3 | 107.3 |
| 200.0 | 191.2 | 95.6 |
| 300.0 | 277.1 | 92.4 |
| 400.0 | 361.5 | 90.4 |
| 500.0 | 490.7 | 98.1 |
| 600.0 | 654.8 | 109.1 |
| 700.0 | 724.4 | 103.5 |
| 800.0 | 872.5 | 109.1 |
| 900.0 | 917.9 | 102.0 |
| 1000.0 | 984.1 | 98.4 |
Analytical Specificity
All compounds tested were added to drug-free, negative human urine and tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes.
Tramadol Metabolites
The cross-reactivity of the following metabolites of tramadol was evaluated by spiking these compounds into drug-free, negative human urine to determine the minimum concentration that would give a positive result approximately equivalent to the 100 ng/mL tramadol cutoff. These concentrations were used to determine the percent cross-reactivity according to the formula:
% Cross-reactivity = (Cutoff concentration / Lowest concentration of cross-reactant causing a positive result) X 100
| Compound | Lowest Concentration
Tested That Produced
a Response
Approximately
Equivalent to the
Cutoff
(ng/mL) | Percent Cross-
reactivity (%) |
|---------------------|-----------------------------------------------------------------------------------------------------------------------|----------------------------------|
| O-Desmethyltramadol | 600 | 16.67 |
| N-Desmethyltramadol | 150 | 66.67 |
8
Structurally Related Compounds
The following structurally related compounds were negative at the concentrations tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes.
| Compound | Concentration Tested (µg/mL) |
|---|---|
| 6-Acetyl Morphine | 10 |
| Amitriptyline | 100 |
| Amphetamine | 100 |
| Chlorpromazine | 50 |
| Clomipramine | 50 |
| Cyclobenzaprine | 10 |
| Desipramine | 50 |
| Dextromethorphan | 100 |
| Diphenhydramine | 500 |
| Doxepin | 50 |
| EDDP | 100 |
| EMDP | 50 |
| Fentanyl | 100 |
| Fluoxetine | 50 |
| Imipramine | 30 |
| Ketamine | 100 |
| MDEA | 75 |
| Meperidine | 100 |
| Methadone | 100 |
| Methapyrilene | 10 |
| Methylphenidate | 100 |
| Methylphenidate Metabolite | 100 |
| Morphine | 50 |
| Morphine-3-glucuronide | 50 |
| N-Desmethyltapentadol | 100 |
| Norcodeine | 100 |
| NorFentanyl | 100 |
| Norketamine | 100 |
| Normeperidine | 50 |
| Normorphine | 50 |
| Noroxycodone | 25 |
| Nortriptyline | 10 |
| Pentazocine (Talwin) | 100 |
| PCP | 100 |
| Propranolol | 15 |
| Quinine | 450 |
| Risperidone | 50 |
| Tapentadol | 100 |
| Thioridazine | 100 |
| Trazodone | 10 |
| Venlafaxine | 100 |
9
Interference – Structurally Unrelated Compounds
High concentrations of the following structurally unrelated compounds were added into urine spiked with tramadol (± 25% of the cutoff concentration) and tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes. The substances listed at the concentrations below did not yield a false result relative to the 100 ng/mL cutoff.
| Compound | Concentration
(ng/mL) | 75 ng/mL
(-25% Cutoff) | 125 ng/mL
(+25% Cutoff) |
|-------------------------------------|--------------------------|---------------------------|----------------------------|
| 6-Acetylcodone | 100,000 | Negative | Positive |
| 6-Acetylmorphine | 100,000 | Negative | Positive |
| 7-Aminoclonazepam | 100,000 | Negative | Positive |
| 7-Aminoflunitrazepam | 100,000 | Negative | Positive |
| 7-Aminonitrazepam | 100,000 | Negative | Positive |
| Albuterol | 100,000 | Negative | Positive |
| Acetaminophen | 500,000 | Negative | Positive |
| Acetylsalicylic Acid | 500,000 | Negative | Positive |
| Alprazolam | 50,000 | Negative | Positive |
| Amitriptyline | 100,000 | Negative | Positive |
| Amobarbital | 100,000 | Negative | Positive |
| S-(+) Amphetamine | 100,000 | Negative | Positive |
| Benzoylecgonine | 500,000 | Negative | Positive |
| Benzylpiperazine | 100,000 | Negative | Positive |
| Bromazepam | 100,000 | Negative | Positive |
| 4-Bromo-2,5,Dimethoxyphenethylamine | 100,000 | Negative | Positive |
| Buprenorphine | 100,000 | Negative | Positive |
| Buprenorphine Glucuronide | 50,000 | Negative | Positive |
| Bupropion | 25,000 | Negative | Positive |
| Butabarbital | 100,000 | Negative | Positive |
| Caffeine | 500,000 | Negative | Positive |
| Cannabidiol | 100,000 | Negative | Positive |
| Cannabinol | 100,000 | Negative | Positive |
| Carbamazepine | 100,000 | Negative | Positive |
| Carisoprodol | 100,000 | Negative | Positive |
| Chlordiazepoxide | 100,000 | Negative | Positive |
| Chlorpromazine | 100,000 | Negative | Positive |
| Clobazam | 100,000 | Negative | Positive |
| Clomipramine | 100,000 | Negative | Positive |
| Clonazepam | 100,000 | Negative | Positive |
| Cocaine | 100,000 | Negative | Positive |
| Codeine | 100,000 | Negative | Positive |
| Cotinine | 100,000 | Negative | Positive |
| Cyclobenzaprine | 100,000 | Negative | Positive |
| Delta-9-THC | 100,000 | Negative | Positive |
| Demoxepam | 100,000 | Negative | Positive |
| Desakylflurazepam | 100,000 | Negative | Positive |
| Desipramine | 100,000 | Negative | Positive |
| Dextromethorphan | 100,000 | Negative | Positive |
| Diazepam | 50,000 | Negative | Positive |
| Compound | Concentration
(ng/mL) | 75 ng/mL
(-25% Cutoff) | 125 ng/mL
(+25% Cutoff) |
| Dihydrocodeine | 100,000 | Negative | Positive |
| Diphenhydramine | 100,000 | Negative | Positive |
| Doxepin | 100,000 | Negative | Positive |
| Ecgonine | 100,000 | Negative | Positive |
| Ecgonine Methyl Ester | 100,000 | Negative | Positive |
| EDDP | 100,000 | Negative | Positive |
| 1R, 2S(-)-Ephedrine | 100,000 | Negative | Positive |
| 1S, 2R(+)-Ephedrine | 100,000 | Negative | Positive |
| EtG | 100,000 | Negative | Positive |
| Ethylmorphine | 100,000 | Negative | Positive |
| R-Fenfluramine | 100,000 | Negative | Positive |
| S-Fenfluramine | 100,000 | Negative | Positive |
| Fentanyl | 100,000 | Negative | Positive |
| Flunitrazepam | 100,000 | Negative | Positive |
| Fluoxetine | 50,000 | Negative | Positive |
| Fluphenazine | 100,000 | Negative | Positive |
| Flurazepam | 100,000 | Negative | Positive |
| Heroin | 100,000 | Negative | Positive |
| Hexobarital | 100,000 | Negative | Positive |
| Hydrocodone | 100,000 | Negative | Positive |
| Hydromorphone | 100,000 | Negative | Positive |
| 11-hydroxy-delta-9-THC | 100,000 | Negative | Positive |
| Ibuprofen | 100,000 | Negative | Positive |
| Imipramine | 100,000 | Negative | Positive |
| Ketamine | 100,000 | Negative | Positive |
| Lamotrigine | 100,000 | Negative | Positive |
| Levorphanol | 75,000 | Negative | Positive |
| Lidocaine | 100,000 | Negative | Positive |
| Lorazepam | 100,000 | Negative | Positive |
| Lorazepam Glucuronide | 50,000 | Negative | Positive |
| Lormetazepam | 100,000 | Negative | Positive |
| LSD | 100,000 | Negative | Positive |
| Maprotiline | 100,000 | Negative | Positive |
| MDA | 100,000 | Negative | Positive |
| MDEA | 10,000 | Negative | Positive |
| MDMA | 50,000 | Negative | Positive |
| Meperidine | 100,000 | Negative | Positive |
| Meprobamate | 100,000 | Negative | Positive |
| Meprotiline | 50,000 | Negative | Positive |
| Methadone | 500,000 | Negative | Positive |
| S(+)-methamphetamine | 500,000 | Negative | Positive |
| Methaqualone | 100,000 | Negative | Positive |
| Methylphenidate | 25,000 | Negative | Positive |
| Metronidazole | 300,000 | Negative | Positive |
| Midazolam | 100,000 | Negative | Positive |
| Morphine | 100,000 | Negative | Positive |
| Morphine-3-beta-glucuronide | 100,000 | Negative | Positive |
| Compound | Concentration
(ng/mL) | 75 ng/mL
(-25% Cutoff) | 125 ng/mL
(+25% Cutoff) |
| Morphine-6-beta-glucuronide | 100,000 | Negative | Positive |
| Nalorphine | 100,000 | Negative | Positive |
| Naloxone | 100,000 | Negative | Positive |
| Naltrexone | 100,000 | Negative | Positive |
| Naproxen | 100,000 | Negative | Positive |
| N-desmethyltapentadol | 25,000 | Negative | Positive |
| Nicotine | 10,000 | Negative | Positive |
| Nitrazepam | 100,000 | Negative | Positive |
| Norbuprenorphine | 100,000 | Negative | Positive |
| Norcodeine | 100,000 | Negative | Positive |
| Nordiazepam | 100,000 | Negative | Positive |
| Normorphine | 100,000 | Negative | Positive |
| Norproproxyphene | 100,000 | Negative | Positive |
| Norpseudoephedrine | 100,000 | Negative | Positive |
| Nortriptyline | 100,000 | Negative | Positive |
| Oxazepam | 100,000 | Negative | Positive |
| Oxazepam Glucuronide | 10,000 | Negative | Positive |
| Oxycodone | 100,000 | Negative | Positive |
| Oxymorphone | 100,000 | Negative | Positive |
| PCP | 10,000 | Negative | Positive |
| Pentazocine | 50,000 | Negative | Positive |
| Pentobarbital | 100,000 | Negative | Positive |
| Phenobarbital | 100,000 | Negative | Positive |
| Phentermine | 100,000 | Negative | Positive |
| Phenylephrine | 100,000 | Negative | Positive |
| Phenylpropanolamine | 100,000 | Negative | Positive |
| Phenytoin | 100,000 | Negative | Positive |
| PMA | 100,000 | Negative | Positive |
| Propoxyphene | 100,000 | Negative | Positive |
| Propranolol | 2,000 | Negative | Positive |
| Protriptyline | 100,000 | Negative | Positive |
| R,R(-)-Pseudoephedrine | 100,000 | Negative | Positive |
| S,S(+)-Pseudoephedrine | 100,000 | Negative | Positive |
| Ranitidine | 100,000 | Negative | Positive |
| Ritalinic Acid | 100,000 | Negative | Positive |
| Salicylic Acid | 100,000 | Negative | Positive |
| Secobarbital | 100,000 | Negative | Positive |
| Sertraline | 50,000 | Negative | Positive |
| Sufentanil Citrate | 10,000 | Negative | Positive |
| Tapentadol | 25,000 | Negative | Positive |
| Temazepam | 100,000 | Negative | Positive |
| 11-nor-9-carboxy THC | 100,000 | Negative | Positive |
| Theophylline | 100,000 | Negative | Positive |
| Thioridazine | 25,000 | Negative | Positive |
| Tilidine | 50,000 | Negative | Positive |
| Trazodone | 100,000 | Negative | Positive |
| Triazolam | 100,000 | Negative | Positive |
| Compound | Concentration
(ng/mL) | 75 ng/mL
(-25% Cutoff) | 125 ng/mL
(+25% Cutoff) |
| Trifluoromethylphenylpiperazine | 100,000 | Negative | Positive |
| Trimipramine | 100,000 | Negative | Positive |
| Valproic Acid | 250,000 | Negative | Positive |
| Zolpidem Tartrate | 100,000 | Negative | Positive |
ARK Diagnostics, Inc. - 510(k) Summary ARK Tramadol Assay
Page 5-7 of 5-12
10
ARK Diagnostics, Inc. – 510(k) Summary ARK Tramadol Assay
Page 5-8 of 5-12
11
ARK Diagnostics, Inc. – 510(k) Summary ARK Tramadol Assay
Page 5-9 of 5-12
12
Interference – Endogenous Substances
Interference studies were performed using CLSI EP07-A2 as a guideline. High concentrations of the following endogenous substances were added into urine spiked with tramadol (± 25% of the cutoff concentration). No interference was observed when tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes.
| Compound | Concentration
Tested | 75 ng/mL
(-25% Cutoff) | 125 ng/mL
(+25% Cutoff) |
|-----------------|-------------------------|---------------------------|----------------------------|
| Acetone | 1000 mg/dL | Negative | Positive |
| Ascorbic Acid | 1500 mg/dL | Negative | Positive |
| Bilirubin | 2 mg/dL | Negative | Positive |
| Creatinine | 500 mg/dL | Negative | Positive |
| Ethanol | 1000 mg/dL | Negative | Positive |
| Galactose | 10 mg/dL | Negative | Positive |
| Gamma Globulin | 500 mg/dL | Negative | Positive |
| Glucose | 3000 mg/dL | Negative | Positive |
| Hemoglobin | 300 mg/dL | Negative | Positive |
| Human Albumin | 500 mg/dL | Negative | Positive |
| Oxalic Acid | 100 mg/dL | Negative | Positive |
| Riboflavin | 7.5 mg/dL | Negative | Positive |
| Sodium Azide | 1% w/v | Negative | Positive |
| Sodium Chloride | 6000 mg/dL | Negative | Positive |
| Sodium Fluoride | 1% w/v | Negative | Positive |
| Urea | 6000 mg/dL | Negative | Positive |
Interference – Specific Gravity and pH
Urine samples with specific gravity values from 1.000 to 1.030 and pH values ranging from 3.0 to 11.0 were tested in the presence of the two levels of tramadol at ± 25% of the cutoff concentration. No interference was observed when tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes.
13
Interference – Boric Acid
One percent (1%) w/v of boric acid was added into urine spiked with tramadol (± 25% of the cutoff concentration) and tested with the ARK Tramadol Assay in both qualitative and semiquantitative modes. Results are provided in the table below.
| Semiquantitative Mode | Qualitative Mode | ||||
|---|---|---|---|---|---|
| Compound | Concentration | ||||
| Tested | 75 ng/mL | ||||
| (-25% Cutoff) | 125 ng/mL | ||||
| (+25% Cutoff) | 75 ng/mL | ||||
| (-25% Cutoff) | 125 ng/mL | ||||
| (+25% Cutoff) | |||||
| Boric Acid | 1% w/v | Negative | Positive | Negative | Negative |
Boric acid interferes with results from this device. Do not test samples that have boric acid as a preservative.
Method Comparison
A total of one hundred fifteen (115) unaltered clinical human urine specimens that are not individually identifiable were analyzed for tramadol with the ARK Tramadol Assay in both qualitative and semiquantitative modes and the results were compared to LC-MS/MS. The LC-MS/MS confirmatory method was performed by a licensed reference laboratory. Results are summarized in the tables below.
| ARK
Immunoassay
Result | Low Negative
Less than
50% below
the Cutoff
( 150 ng/mL
by LC-
MS/MS) |
|------------------------------|-----------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Negative | 50 | 0 | 0 | 0 |
| Positive | 0 | 5* | 4 | 56 |
*Discordant Results
| Sample ID Number | ARK Immunoassay
Result | Tramadol
(ng/mL by LC-MS/MS) |
|------------------|---------------------------|---------------------------------|
| 01 | Positive | 74.0 |
| 05 | Positive | 98.7 |
| 06 | Positive | 98.9 |
| 51 | Positive | 75.0 |
| 52 | Positive | 79.0 |
O-desmethyltramadol was detected in these samples and contributed to the positive result obtained with the ARK Tramadol Assay.
14
Traceability and Value Assignment
ARK Tramadol Calibrators and Controls are prepared by volumetric dilution of high purity tramadol (certified solution traceable to HPLC) into non-sterile, processed human urine free of tramadol. Testing is performed with the ARK Tramadol Assay on the Beckman Coulter AU680 automated clinical chemistry analyzer, calibrated with the ARK Tramadol Calibrator.
Calibration Curve Stability
A stored calibration curve was effective up to at least 30 days based on supporting data. Calibration curve stability may depend on individual laboratory performance.
807.92 (b)(3): Conclusions from Nonclinical Testing
As summarized above, the ARK Tramadol Assay is substantially equivalent to the legally marketed predicate device, Immunalysis Tramadol Urine Enzyme Immunoassay (K141803).