← Product Code [DJG](/productcode/DJG) · K141803

# IMMUNALYSIS TRAMADOL URINE ENZYME IMMUNOASSAY (K141803)

_Immunalysis Corporation · DJG · Feb 12, 2015 · Clinical Toxicology · SESE_

**Canonical URL:** https://fda.innolitics.com/device/K141803

## Device Facts

- **Applicant:** Immunalysis Corporation
- **Product Code:** [DJG](/productcode/DJG.md)
- **Decision Date:** Feb 12, 2015
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 862.3650
- **Device Class:** Class 2
- **Review Panel:** Clinical Toxicology
- **Attributes:** Real-World Evidence

## Real-World Evidence

| Submission | Device | Sponsor | RWD Sources | RWE Use Summary | Key Tags |
| --- | --- | --- | --- | --- | --- |
| K141803 · Feb 12, 2015 | IMMUNALYSIS TRAMADOL URINE ENZYME IMMUNOASSAY | Immunalysis Corporation | Discarded clinical urine samples from clinical testing laboratories | The sponsor used discarded clinical urine samples to perform a method comparison study, evaluating the performance of the Immunalysis Tramadol Urine Enzyme Immunoassay against LC/MS confirmation. | Method comparison; Clinical urine samples; Discarded samples; Analytical validation |

### Clinical Evidence

| Study Design | Population | Comparator | Key Endpoints |
| --- | --- | --- | --- |
| Method comparison study using clinical samples | Clinical urine samples; Sample Size: 150 samples (100 positive, 50 negative based on LC/MS) | Liquid Chromatography Tandem Mass Spectrometry (LC/MS) | Qualitative and semi-quantitative agreement with LC/MS |

## Indications for Use

The Immunalysis Tramadol Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay with a cutoff of 200 ng/mL. The assay is intended for use in laboratories for the qualitative and semi-quantitative analysis of Tramadol in human urine with automated clinical chemistry analyzers. This assay is calibrated against Tramadol. This in-vitro diagnostic device is for prescription use only. The semi-quantitative mode is for purposes of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GC-MS or permitting laboratories to establish quality control procedures. The test is not intended to differentiate between drugs of abuse and prescription use of Tramadol. There are no uniformly recognized drug levels for Tramadol in urine. The Immunalysis Tramadol Urine Enzyme Immunoassay Kit provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography / Mass Spectrometry (GC/MS) or Liquid Chromatography / Mass Spectroscopy (LC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used. The Immunalysis Tramadol Urine Controls are used as control materials in the Immunalysis Tramadol Urine Enzyme Immunoassay. The Immunalysis Tramadol Urine Calibrators are used as calibrators in the Immunalysis Tramadol Urine Enzyme Immunoassay for the qualitative and semi-quantitative determination of Tramadol in urine on automated clinical chemistry analyzers.

## Device Story

Homogeneous enzyme immunoassay for detection of Tramadol in human urine; utilizes goat polyclonal antibodies to Tramadol; competitive binding mechanism between drug in sample and enzyme-labeled drug conjugate (G6PDH) for fixed antibody sites; enzyme activity measured spectrophotometrically at 340 nm via NAD to NADH conversion; inverse relationship between drug concentration and enzyme activity. Used in clinical laboratories on automated chemistry analyzers; operated by laboratory technicians. Provides preliminary qualitative/semi-quantitative results; requires confirmation by GC-MS or LC/MS. Assists clinicians in identifying potential Tramadol presence to guide further diagnostic or confirmatory testing.

## Clinical Evidence

Bench testing only. Performance evaluated using 150 clinical urine samples compared against LC/MS confirmation. Qualitative and semi-quantitative analysis showed 100% agreement with LC/MS. Precision study (N=80) performed over 20 days. Specificity/cross-reactivity and interference studies conducted; Boric Acid identified as an interferent. Linearity verified in semi-quantitative mode. Stability studies (accelerated and open-vial) support 1-year shelf life and 28-day on-board stability.

## Technological Characteristics

Homogeneous enzyme immunoassay; reagents include goat anti-Tramadol antibodies, G6P, NAD, and G6PDH-labeled Tramadol conjugate. Liquid, ready-to-use format. Analyzed via spectrophotometry at 340 nm. Requires automated clinical chemistry analyzer capable of temperature control, pipetting, and kinetic rate measurement. Storage at 2-8°C. No specific ASTM material standards cited.

## 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

- LZI Opiate 2000 Enzyme Immunoassay (k120761)

## Submission Summary (Full Text)

> This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com.
>
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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
k141803

B. Purpose for Submission:
New device

C. Measurand:
Tramadol

D. Type of Test:
Homogenous Enzyme Immunoassay, Qualitative and Semi-quantitative.

E. Applicant:
Immunalysis Corporation

F. Proprietary and Established Names:
Immunalysis Tramadol Urine Enzyme Immunoassay
Immunalysis Tramadol Urine Controls
Immunalysis Tramadol Urine Calibrators

G. Regulatory Information:

1. Regulation section:
21 CFR 862.3650, Opiate Test System
21 CFR 862.3200, Clinical Toxicology Calibrator
21 CFR 862.3280, Clinical Toxicology Control Materials

2. Classification:
Class II
Class I, reserved.

3. Product code:
DJG
DLJ
LAS

4. Panel:
Toxicology (91)

H. Intended Use:

1. Intended use(s):
See Indications for use below.

2. Indication(s) for use:

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The Immunalysis Tramadol Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay with a cutoff of 200 ng/mL. The assay is intended for use in laboratories for the qualitative and semi-quantitative analysis of Tramadol in human urine with automated clinical chemistry analyzers. This assay is calibrated against Tramadol. This in-vitro diagnostic device is for prescription use only.

The semi-quantitative mode is for purposes of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GC-MS or permitting laboratories to establish quality control procedures. The test is not intended to differentiate between drugs of abuse and prescription use of Tramadol. There are no uniformly recognized drug levels for Tramadol in urine.

The Immunalysis Tramadol Urine Enzyme Immunoassay Kit provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography / Mass Spectrometry (GC/MS) or Liquid Chromatography / Mass Spectroscopy (LC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

The Immunalysis Tramadol Urine Controls are used as control materials in the Immunalysis Tramadol Urine Enzyme Immunoassay.

The Immunalysis Tramadol Urine Calibrators are used as calibrators in the Immunalysis Tramadol Urine Enzyme Immunoassay for the qualitative and semi-quantitative determination of Tramadol in urine on automated clinical chemistry analyzers.

3. Special conditions for use statement(s):

For prescription use only.

Not for use in Point of Care settings.

4. Special instrument requirements:

Beckman Coulter AU 400e chemistry analyzer was used to generate data for this submission.

Automated clinical chemistry analyzers capable of maintaining a constant reaction temperature, pipetting samples and reagents, mixing reagents, timing reactions and measuring enzyme rates at 340 nm can be used for the assay.

I. Device Description:

Immunalysis Tramadol Urine Enzyme Immunoassay (EIA) Kit includes antibody/ substrate reagent and enzyme conjugate reagent.

- Antibody/ substrate reagent includes goat antibodies to Tramadol, glucose-6-phosphate (G6P) and nicotinamide adenine dinucleotide (NAD) in Tris buffer with Sodium Azide as a preservative.
- Enzyme conjugate reagent includes tramadol derivative labeled with glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with Sodium Azide as a preservative.

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Calibrators and controls are sold separately. The tramadol calibrators and controls consist of:

- A single calibrator - 200ng/mL Tramadol.
- A control set - contains a LOW control at 150ng/mL and a HIGH control at 250ng/mL.
- A calibrator set - contains a negative calibrator, a Level 1 calibrator at 100ng/mL, a Level 2 calibrator at 200ng/mL, a Level 3 calibrator at 500ng/mL and a Level 4 calibrator at 1000ng/mL tramadol.

All reagents are in liquid form and ready to use.

J. Substantial Equivalence Information:

1. Predicate device name(s):
LZI Opiate 2000 Enzyme Immunoassay
LZI Opiate 2000 Enzyme Controls
LZI Opiate 2000 Enzyme Calibrators

2. Predicate 510(k) number(s):
k120761

3. Comparison with predicate:

|  Item | Immunalysis Tramadol Urine Enzyme Immunoassay (Candidate Device) | LZI Opiate 2000 Enzyme Immunoassay, k120761 (Predicate Device)  |
| --- | --- | --- |
|  Intended use | For the qualitative and semi-quantitative determination of the presence of drugs of abuse in human urine. For in vitro diagnostic use. | Same  |
|  Measured analyte(s) | Tramadol | Opiates  |
|  Assay cutoff | 200 ng/mL of Tramadol | 2000 ng/mL of Opiates  |
|  Assay calibrated against | Tramadol | Morphine  |
|  Test system type | Homogenous enzyme immunoassay | Enzyme immunoassay  |
|  Storage conditions | 2 - 8°C until expiration date | Same  |
|  Calibrator form | Liquid | Same  |
|  Calibrator levels | One level (200 ng/mL) | One level (2000 ng/mL)  |
|  Control set levels | Two levels (150 ng/mL and 250ng/mL) | Two levels (1500 ng/mL and 2500 ng/mL)  |

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|  Item | Immunalysis Tramadol Urine Enzyme Immunoassay (Candidate Device) | LZI Opiate 2000 Enzyme Immunoassay, k120761 (Predicate Device)  |
| --- | --- | --- |
|  Calibrator set levels | Five levels (0, 100, 200, 500 and 1000 ng/mL) | Five levels (0, 1000, 2000, 4000 and 6000 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

L. Test Principle:

The assay is based on the competition of Tramadol labeled enzyme glucose-6-phosphate dehydrogenase (G6PDH) and the free Tramadol in the urine sample for the fixed amount of antibody binding sites. In the absence of the free Tramadol in the urine sample, the antibody (goat-polyclonal) binds the Tramadol enzyme conjugate and enzyme activity is inhibited. This creates a dose response relationship between Tramadol concentration in the urine and enzyme activity. The enzyme G6PDH activity is determined at 340 nm spectrophotometrically by the conversion of NAD to NADH.

M. Performance Characteristics (if/when applicable):

All the performance studies were performed on the Beckman Coulter AU 400e chemistry analyzer.

1. Analytical performance:

a. Precision/Reproducibility:

A precision study was performed by one experienced operator following the CLSI (EP5-A2) precision guidelines. Negative urine sample pools were spiked with Tramadol at 0, 50, 100, 150, 200, 250, 300, 350, and 400 ng/mL, representing 0, 25, 50, 75, 100, 125, 150, 175 and 200% of the device cutoff (200 ng/mL). Each level sample was tested in duplicate per run, two runs per day for twenty consecutive days (total N= 80/level) on the Beckman Coulter AU 400e chemistry analyzer. The results are summarized in the table below:

Qualitative Precision Result:

|  Concentration as % of the Cutoff Level | Target Tramadol concentration (ng/mL) | Immunalysis Tramadol Urine EIA # Neg / # Pos  |
| --- | --- | --- |
|  0 | 0 | 80 Neg / 0 Pos  |
|  25 | 50 | 80 Neg / 0 Pos  |
|  50 | 100 | 80 Neg / 0 Pos  |

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|  75 | 150 | 80 Neg / 0 Pos  |
| --- | --- | --- |
|  100 | 200 | 44 Neg / 36 Pos  |
|  125 | 250 | 0 Neg / 80 Pos  |
|  150 | 300 | 0 Neg / 80 Pos  |
|  175 | 350 | 0 Neg / 80 Pos  |
|  200 | 400 | 0 Neg / 80 Pos  |

## Semi-Quantitative Precision Result:

|  Concentration as % of the Cutoff Level | Target Tramadol concentration (ng/mL) | Immunalysis Tramadol Urine EIA # Neg / # Pos  |
| --- | --- | --- |
|  0 | 0.00 | 80 Neg / 0 Pos  |
|  25 | 50 | 80 Neg / 0 Pos  |
|  50 | 100 | 80 Neg / 0 Pos  |
|  75 | 150 | 80 Neg / 0 Pos  |
|  100 | 200 | 47 Neg / 33 Pos  |
|  125 | 250 | 0 Neg / 80 Pos  |
|  150 | 300 | 0 Neg / 80 Pos  |
|  175 | 350 | 0 Neg / 80 Pos  |
|  200 | 400 | 0 Neg / 80 Pos  |

## b. Linearity/assay reportable range:

Linearity study in the semi-quantitative mode was conducted by spiking drug free urine pool with tramadol (serial dilutions of a high concentration tramadol in urine in increments of 10%) to achieve concentrations ranging from 100ng/mL to 1100ng/mL, and testing each level in duplicate on the Beckman Coulter AU 400e clinical chemistry analyzer. The results are summarized below:

|  Expected Concentration (ng/mL) | Mean Concentration (ng/mL) | Recovery (%)  |
| --- | --- | --- |
|  0 | 1 | N/A  |
|  100 | 102 | 102  |
|  200 | 196 | 98  |
|  300 | 323 | 108  |

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|  Expected Concentration (ng/mL) | Mean Concentration (ng/mL) | Recovery (%)  |
| --- | --- | --- |
|  |   |   |
|  400 | 416 | 104  |
|  500 | 506 | 101  |
|  600 | 558 | 93  |
|  700 | 637 | 91  |
|  800 | 756 | 95  |
|  900 | 872 | 97  |
|  1000 | 956 | 96  |
|  1100 | 1012 | 92  |

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

## Traceability

Calibrators and controls are prepared from a standard solution of tramadol purchased from a commercial vendor. This standard solution is diluted with a BSA buffer to make the cutoff calibrator, control set of low and high controls, and calibrator set (of 5 level calibrators). The concentrations of the prepared solutions are confirmed by GC/MS or LC/MS/MS.

## Value Assignment – Calibrators and Controls

A commercially available standard solution of Tramadol is mixed with a commercially available BSA buffer to the desired calculated concentrations for the LOW control, HIGH control and Calibrators. The calibrators and controls are tested by GC/MS or LC/MS/MS. Values are assigned to the controls once the GC/MS or LC/MS/MS results are within acceptable ranges. The negative standard is prepared with the BSA buffer. The negative standard is compared to a reference negative standard to ensure that it is free of tramadol. Value is assigned when the test result is within the acceptable range. Acceptance criteria for value assignment were reviewed and deemed acceptable.

## Calibrators and Controls Stability Studies

Accelerated and real-time stability studies in the qualitative and semi-quantitative modes were conducted on multiple lots of Immunalysis Tramadol Urine Controls and Immunalysis Tramadol Urine Calibrators. The stability protocols and acceptance criteria for open and closed vial were reviewed and found acceptable. The open vial

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and closed vial study results support the open vial stability claim of six months and closed vial stability claim of twelve months when stored at 2 to 8 °C for the Immunalysis Tramadol Urine Controls and Immunalysis Tramadol Urine Calibrators.

## Sample Storage - Stability Studies

A specimen storage study was performed using urine specimens at tramadol concentration of 147 ng/mL (below the cutoff) and 245 ng/mL (above the cutoff) to establish specimen storage stability at 2 – 8°C. The urine specimen at tramadol concentration below the cutoff was negative in comparison to the 200ng/mL cutoff for day 0, week 1, 2, 3 and 4. The urine specimen at tramadol concentration above the cutoff was positive in comparison to the 200ng/mL cutoff for Day 0, Week 1, 2, 3 and 4. The stability study supported a one month urine specimen containing tramadol stability when stored at 2 – 8°C.

## d. Detection limit:

Not applicable.

## e. Analytical specificity:

An analytical specificity study to evaluate interference from non-structurally and structurally related compounds was performed in the qualitative and semi-quantitative mode. The study design and results are described below. Results were the same for each mode (qualitative and semi-quantitative modes).

## Structurally Related Compounds

Compounds sharing structural or conformational similarity to tramadol were tested for cross-reactivity with the candidate device. The structurally related compounds that exhibited cross-reactivity with the candidate device in urine were titrated to determine the percent cross-reactivity. The concentration (ng/mL) of cross-reactant that gives a response equivalent to the cutoff, and the percent cross-reactivity are presented in the table below.

|  Compound | Concentration of the compound tested (ng/mL) | Immunoassay Result | Percent Cross-reactivity  |
| --- | --- | --- | --- |
|  Tramadol | 200 | Positive | 100.00  |
|  n-Desmethyl Tramadol | 450 | Positive | 44.40  |
|  o-Desmethyl Tramadol | 25,000 | Positive | 0.80  |
|  Venlafaxine | 100,000 | Negative | Not detected (< 0.05%)  |
|  o-Desmethyl Venlafaxine | 100,000 | Negative | Not detected (< 0.05%)  |

## Non-Structurally Related Compounds

Potential interference from non-structurally related drugs and metabolites was

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evaluated in the qualitative and semi-quantitative modes, by spiking these compounds at high concentrations in pooled urine spiked with tramadol at  $\pm 25\%$  of the cutoff (150 and  $250~\mathrm{ng / mL}$ ).

|  Compounds | Concentration tested (ng/mL) | -25% Cutoff (150ng/mL) | +25% Cutoff (250ng/mL)  |
| --- | --- | --- | --- |
|  6-Acetylcodeine | 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  |
|  Acetaminophen | 500,000 | Negative | Positive  |
|  Acetylsalicyclic 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  |
|  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  |
|  Dexamethasone | 100,000 | Negative | Positive  |
|  Dexylsalicylic acid | 100,000 | Negative | Positive  |
|  Dexylphenethylamine | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |
|  Dexylphenidate | 100,000 | Negative | Positive  |

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|  Dihydrocodeine | 100,000 | Negative | Positive  |
| --- | --- | --- | --- |
|  Diphenhydramine | 500,000 | 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  |
|  Fenfluramine | 100,000 | Negative | Positive  |
|  Fentanyl | 100,000 | Negative | Positive  |
|  Flunitrazepam | 100,000 | Negative | Positive  |
|  Fluoxetine | 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 | 100,000 | 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  |
|  S(+)-MDA | 100,000 | Negative | Positive  |
|  MDEA | 100,000 | Negative | Positive  |
|  MDMA | 100,000 | Negative | Positive  |
|  Meperidine | 100,000 | Negative | Positive  |
|  Meprobamate | 100,000 | Negative | Positive  |
|  Methadone | 500,000 | Negative | Positive  |
|  S(+)-Methamphetamine | 500,000 | Negative | Positive  |
|  Methaquolone | 100,000 | Negative | Positive  |
|  Methylphenidate | 100,000 | Negative | Positive  |
|  Midazolam | 100,000 | Negative | Positive  |
|  Morphine | 100,000 | Negative | Positive  |
|  Morphine-3 -glucuronide | 100,000 | Negative | Positive  |
|  Morphine-6 -glucuronide | 100,000 | Negative | Positive  |
|  Nalorphine | 100,000 | Negative | Positive  |

9

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|  Naloxone | 100,000 | Negative | Positive  |
| --- | --- | --- | --- |
|  Naltrexone | 100,000 | Negative | Positive  |
|  Naproxen | 100,000 | Negative | Positive  |
|  N-desmethyltapentadol | 100,000 | 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 | 100,000 | Negative | Positive  |
|  Pentazocine | 100,000 | 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 | 100,000 | 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 | 100,000 | Negative | Positive  |
|  Sufentanil Citrate | 100,000 | Negative | Positive  |
|  Temazepam | 100,000 | Negative | Positive  |
|  11-nor-9 carboxy THC | 100,000 | Negative | Positive  |
|  Theophylline | 100,000 | Negative | Positive  |
|  Thioridazine | 100,000 | Negative | Positive  |
|  Trazodone | 100,000 | Negative | Positive  |
|  Triazolam | 100,000 | Negative | Positive  |
|  Trifluoromethylphenyl-piperazine | 100,000 | Negative | Positive  |
|  Trimipramine | 100,000 | Negative | Positive  |

10

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Zolpidem Tartrate 100,000 Negative Positive

# Endogenous Compounds

Potential interference from endogenous compounds was evaluated in the qualitative and semi-quantitative modes, by spiking these compounds at high concentrations in pooled negative urine and in pooled urine spiked with tramadol at  $\pm 25\%$  of the cutoff (150 and  $250~\mathrm{ng / mL}$ ).

|  Compounds | Concentration tested | -25% Cutoff (150ng/mL) | +25% Cutoff (250ng/mL)  |
| --- | --- | --- | --- |
|  Acetone | 1.0 g/dL | Negative | Positive  |
|  Ascorbic Acid | 1.5 g/dL | Negative | Positive  |
|  Bilirubin | 0.002 g/dL | Negative | Positive  |
|  Boric Acid | 1% w/v | Negative | Negative  |
|  Creatinine | 0.5 g/dL | Negative | Positive  |
|  Ethanol | 1.0 g/dL | Negative | Positive  |
|  Galactose | 0.01 g/dL | Negative | Positive  |
|  γ-Globulin | 0.5 g/dL | Negative | Positive  |
|  Glucose | 2.0 g/dL | Negative | Positive  |
|  Hemoglobin | 0.300 g/dL | Negative | Positive  |
|  Human Serum Albumin | 0.5 g/dL | Negative | Positive  |
|  Oxalic Acid | 0.1 g/dL | Negative | Positive  |
|  Riboflavin | 0.0075 g/dL | Negative | Positive  |
|  Sodium Azide | 1% w/v | Negative | Positive  |
|  Sodium Chloride | 6.0 g/dL | Negative | Positive  |
|  Sodium Flouride | 1% w/v | Negative | Positive  |
|  Urea | 6.0 g/dL | Negative | Positive  |

Compounds that showed interference was further evaluated by spiking it in pooled urine spiked with tramadol at  $\pm 50\%$  of the cutoff (100 and  $300~\mathrm{ng / mL}$ ). Boric Acid at  $1\%$  w/v caused a false negative response for tramadol at  $+25\%$  and also at  $+50\%$  of the cutoff.

The following statement is listed under the Limitations section of the labeling, "Boric Acid at  $1\%$  w/v may cause false negative results. Boric Acid is not recommended as a preservative for urine."

# 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 urine containing Tramadol at  $\pm 25\%$  of the cutoff (150 ng/mL and 250 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

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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  $\pm 25\%$  of the cutoff (150 ng/mL and 250 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:

Characterization of how the device performs analytically around the claimed cutoff concentration of  $200\mathrm{ng / mL}$  tramadol is described in the precision section, M.1.a. above.

2. Comparison studies:

a. Method comparison with predicate device:

A total 150 unaltered urine samples from clinical testing laboratories were analyzed by the candidate device in the qualitative and semi-quantitative modes on the Beckman Coulter AU 400e clinical chemistry analyzer and the comparative mass spectrometry based quantitative method (LC/MS). The results from the study are summarized below:

Qualitative Mode

|  Candidate Device Results | <50% of cutoff concentration by LC/MS (< 100ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC/MS) (100 ~ 199 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC/MS) (200 ~ 300 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC/MS) > 300 ng/mL  |
| --- | --- | --- | --- | --- |
|  Positive | 0 | 0 | 10 | 90  |
|  Negative | 45 | 5 | 0 | 0  |

$\%$  Agreement among positives is  $100\%$ .
$\%$  Agreement among negatives is  $100\%$ .

Semi-quantitative Mode

|  Candidate Device Results | <50% of cutoff concentration by LC/MS (< 100ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC/MS) (100 ~ 199 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC/MS) (200 ~ 300 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC/MS) > 300 ng/mL  |
| --- | --- | --- | --- | --- |
|  Positive | 0 | 0 | 10 | 90  |

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|  Negative | 45 | 5 | 0 | 0  |
| --- | --- | --- | --- | --- |

% Agreement among positives is 100%.

% Agreement among negatives is 100%.

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):

Literature was provided to support a screening cut-off of 200 ng/mL for tramadol in urine.

Melanson, S.E.F., Ptolemy, A.S., and Wasan, A.D. Optimizing Urine Drug Testing for Monitoring Medication Compliance in Pain Management. Pain Medicine 2013; 14: 1813 -1820.

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 Part 809.10.

O. Conclusion:

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

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**Source:** [https://fda.innolitics.com/device/K141803](https://fda.innolitics.com/device/K141803)

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