← Product Code [DJG](/productcode/DJG) · K182280

# ARK Tramadol Assay (K182280)

_Ark Diagnostics, Inc. · DJG · Dec 10, 2018 · Clinical Toxicology · SESE_

**Canonical URL:** https://fda-staging.innolitics.com/device/K182280

## Device Facts

- **Applicant:** Ark Diagnostics, Inc.
- **Product Code:** [DJG](/productcode/DJG.md)
- **Decision Date:** Dec 10, 2018
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 862.3650
- **Device Class:** Class 2
- **Review Panel:** Clinical Toxicology

## 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.

## Device Story

ARK Tramadol Assay is a homogeneous enzyme immunoassay for detecting tramadol in human urine; used in clinical laboratories on automated chemistry analyzers (e.g., Beckman Coulter AU 680). Input: human urine specimen. Principle: competitive binding between drug in specimen and tramadol-labeled recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites; enzyme activity increases proportionally to free tramadol concentration. Output: absorbance change measured spectrophotometrically at 340 nm, indicating presence/concentration of tramadol. Results are preliminary; require confirmation via GC/MS or LC-MS/MS. Clinical utility: aids in identifying tramadol presence for clinical decision-making; enables dilution determination for confirmatory testing and quality control. Benefits: provides rapid preliminary screening for tramadol in urine.

## Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP05-A3 (N=160 per level). Linearity assessed from 50-1000 ng/mL. Analytical specificity tested against numerous structurally related and unrelated compounds; cross-reactivity reported for O-desmethyltramadol (16.67%) and N-desmethyltramadol (66.67%). Interference testing confirmed no false results with common endogenous substances or high-concentration drugs, excluding boric acid. Method comparison performed on 115 clinical urine specimens against LC-MS/MS reference method; results showed concordance with minor discordance attributed to metabolite cross-reactivity.

## Technological Characteristics

Homogeneous enzyme immunoassay. Reagents: R1 (rabbit polyclonal antibodies, glucose-6-phosphate, NAD, stabilizers) and R2 (tramadol-rG6PDH conjugate). Detection: spectrophotometric absorbance change at 340 nm. Form factor: liquid, ready-to-use. Connectivity: integrated with automated clinical chemistry analyzers (e.g., Beckman Coulter AU 680). Storage: 2-8°C.

## Regulatory 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.

## Special Controls

*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).

## Predicate Devices

- Immunalysis Tramadol Urine Enzyme Immunoassay (k141803)

## Submission Summary (Full Text)

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>
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1

510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
k182280

B. Purpose for Submission:
New device

C. Measurand:
Tramadol

D. Type of Test:
Homogenous enzyme immunoassay, qualitative and semi-quantitative

E. Applicant:
ARK Diagnostics, Inc.

F. Proprietary and Established Names:
ARK Tramadol Assay

G. Regulatory Information:
1. Regulation section:
21 CFR 862.3650, Opiate test system
2. Classification:
Class II
3. Product code:
DJG
4. Panel:
Toxicology (91)

H. Intended Use:
1. Intended use(s):
See Indications for use below.
2. Indication(s) 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

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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.

3. Special conditions for use statement(s):

For prescription use only.

4. Special instrument requirements:

Beckman Coulter AU 680 chemistry analyzer

I. Device Description:

The ARK Tramadol Assay is a homogeneous enzyme immunoassay used for the detection of tramadol in human urine. ARK Tramadol Assay includes the following reagents:

Reagent R1 – Antibody/Substrate

Rabbit polyclonal antibodies to tramadol, glucose-6-phosphate, nicotinamide adenine dinucleotide, bovine serum albumin, sodium azide, and stabilizers

Reagent R2 – Enzyme

Tramadol derivative labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH), bovine serum albumin, buffer, sodium azide and stabilizers

All reagents are in liquid form and ready to use.

J. Substantial Equivalence Information:

1. Predicate device name(s):

Immunalysis Tramadol Urine Enzyme Immunoassay

2. Predicate 510(k) number(s):

k141803

3. Comparison with predicate:

|  Characteristic | Predicate Device Immunalysis Tramadol Urine Enzyme Immunoassay (k141803) | Candidate Device ARK Tramadol Assay  |
| --- | --- | --- |
|  Similarities  |   |   |
|  Intended Use | For the qualitative and semiquantitative determination of tramadol in human urine; For in vitro diagnostic use | Same  |

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|  Characteristic | Predicate Device Immunalysis Tramadol Urine Enzyme Immunoassay (k141803) | Candidate Device ARK Tramadol Assay  |
| --- | --- | --- |
|  Sample Matrix | Human urine | Same  |
|  Test System | Homogenous enzyme immunoassay (EIA) | Same  |
|  User Environment | Clinical laboratories; Prescription use only | Same  |
|  Reagents Form | Liquid – Ready to use | Same  |
|  Reagent Materials | Two reagent system: Antibody/substrate reagent and enzyme labeled conjugate Sodium azide preservative | Same  |
|  Storage | 2-8°C | 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  |   |   |
|  Cutoff Level | 200 ng/mL | 100 ng/mL  |

# K. Standard/Guidance Document Referenced (if applicable):

- CLSI EP5-A2: Evaluation of Precision Performance of Quantitative Measurement Methods.
- CLSI EP7-A2: Interference Testing in Clinical Chemistry
- CLSI EP12-A2: User Protocol for Evaluation of Qualitative Test Performance

# L. Test Principle:

The ARK Tramadol Assay is based on competition between drug in the specimen and drug labeled with recombinant glucose-6-phosphate dehydrogenase (tramadol-rG6PDH), for antibody binding sites. When tramadol-rG6PDH is bound by antibody, enzyme activity decreases. In the presence of free tramadol from the specimen, the free tramadol competes for antibody binding to tramadol-rG6PDH, resulting in increased enzyme activity that is proportional to the free tramadol concentration. Active tramadol-rG6PDH converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate

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(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.

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

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 Mode:**

|  Human Urine (ng/mL) | % Cutoff | # of Determination | Qualitative Precision  |
| --- | --- | --- | --- |
|  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 Mode:**

|  Human Urine (ng/mL) | % Cutoff | # of Determinations | Semiquantitative 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 | 97 Negative/ 63 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  |

#### b. Linearity/assay reportable range:

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

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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  |

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

ARK Tramadol Calibrators and Controls are traceable to high purity certified commercial tramadol.

d. Detection limit:

Not applicable.

e. Analytical specificity:

Cross-reactivity

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 yield 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

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|  Compound | Lowest Concentration Tested That Produced a Response Approximately Equivalent to the Cutoff (ng/mL) | Percent Cross-reactivity (%)  |
| --- | --- | --- |
|  Tramadol | 100 | 100  |
|  O-Desmethyltramadol | 600 | 16.67  |
|  N-Desmethyltramadol | 150 | 66.67  |

The following structurally related compounds do not cross-react with the ARK Tramadol Assay in both qualitative and semiquantitative modes at the concentration tested:

|  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-Desmethyltentandol | 100  |
|  Norcodeine | 100  |
|  NorFentanyl | 100  |
|  Norketamine | 100  |

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7

|  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  |

## Interference

High concentrations of the structurally unrelated compounds or endogenous substances 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-Acetylcodine | 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- | 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  |

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|  Compound | Concentration (ng/mL) | 75 ng/mL (-25% Cutoff) | 125 ng/mL (+25% Cutoff)  |
| --- | --- | --- | --- |
|  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  |
|  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  |
|  Hexobarbital | 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  |

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|  Compound | Concentration (ng/mL) | 75 ng/mL (-25% Cutoff) | 125 ng/mL (+25% Cutoff)  |
| --- | --- | --- | --- |
|  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  |
|  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  |
|  Norpropoxyphene | 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  |

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|  Compound | Concentration (ng/mL) | 75 ng/mL (-25% Cutoff) | 125 ng/mL (+25% Cutoff)  |
| --- | --- | --- | --- |
|  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  |
|  Trifluoromethylphenylpiperazine | 100,000 | Negative | Positive  |
|  Trimipramine | 100,000 | Negative | Positive  |
|  Valproic Acid | 250,000 | Negative | Positive  |
|  Zolpidem Tartrate | 100,000 | Negative | Positive  |
|  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  |

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|  Endogenous Substances | 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  |

## 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.

|  Compound | Concentration Tested | Semiquantitative Mode |   | Qualitative Mode  |   |
| --- | --- | --- | --- | --- | --- |
|   |   |  75 ng/mL (-25% Cutoff) | 125 ng/mL (+25% Cutoff) | 75 ng/mL (-25% Cutoff) | 125 ng/mL (+25% Cutoff)  |
|  Boric | 1% w/v | Negative | Positive | Negative | Negative  |

The following statement is listed under the Limitations section of the labeling, "Do not use Boric Acid as a preservative."

## pH and Specific Gravity

For potential interference from the pH of urine, device performance in the qualitative and semi-quantitative modes was tested using a range of urine pH values (3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 and 11.0). All test samples were prepared in drug free

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urine containing Tramadol at ± 25% of the cutoff (75 ng/mL and 125 ng/mL tramadol concentrations). No positive or negative interference was observed at urine pH values ranging from 3.0 to 11.0 for each test mode.

For potential interference from the specific gravity of urine, device performance in the qualitative and semi-quantitative modes was tested using a range of urine specific gravity values (1.000, 1.002, 1.005, 1.010, 1.015, 1.020, 1.025 and 1.030). All test samples were prepared in drug free urine containing Tramadol at ± 25% of the cutoff (75 ng/mL and 125 ng/mL tramadol concentrations). No positive or negative interference was observed at urine specific gravity values ranging from 1.000 to 1.030 for each test mode.

f. Assay cut-off:

Not applicable.

2. Comparison studies:

a. Method comparison with predicate device:

A total of 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 (< 50) | Near Cutoff Negative Between 50% below the Cutoff and the Cutoff (50 – 99) | Near Cutoff Positive Between the Cutoff and 50% above the Cutoff | High Positive Greater than 50% above the Cutoff  |
| --- | --- | --- | --- | --- |
|  Negative | 50 | 0 | 0 | 0  |
|  Positive | 0 | 5* | 4 | 56  |

*Discordant Results: O-desmethyltramadol was detected in the below samples and contributed to the positive result obtained with the ARK Tramadol Assay.

|  Discordant Sample ID # | Tramadol by LC/MS- MS (ng/mL) | Candidate Device (Positive/Negative)  |
| --- | --- | --- |
|  01 | 74 | Positive  |
|  05 | 98.7 | Positive  |
|  06 | 98.9 | Positive  |

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|  Discordant Sample ID # | Tramadol by LC/MS- MS (ng/mL) | Candidate Device (Positive/Negative)  |
| --- | --- | --- |
|  51 | 75.0 | Positive  |
|  52 | 79.0 | Positive  |

b. Matrix comparison:
Not applicable. Urine is the only claimed matrix for the candidate device.

3. Clinical studies:
a. Clinical Sensitivity:
Not applicable.
b. Clinical specificity:
Not applicable.
c. Other clinical supportive data (when a. and b. are not applicable):
The sponsor provided published, peer-reviewed articles and reports which, in addition to data provided by the sponsor, supported that for doses of tramadol anticipated in the intended use population, a 100 ng/mL cutoff would detect the analyte for approximately 3-4 days (60-100 hours).

4. Clinical cut-off:
Not applicable.

5. Expected values/Reference range:
Not applicable.

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Parts 801 and 809, as applicable.

O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

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**Source:** [https://fda-staging.innolitics.com/device/K182280](https://fda-staging.innolitics.com/device/K182280)

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