RANDOX METHADONE ASSAY

K092272 · Randox Laboratories, Ltd. · DJR · Nov 18, 2010 · Clinical Toxicology

Device Facts

• Record ID: K092272
• Device Name: RANDOX METHADONE ASSAY
• Applicant: Randox Laboratories, Ltd.
• Product Code: DJR · Clinical Toxicology
• Decision Date: Nov 18, 2010
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3620
• Device Class: Class 2

Indications for Use

The Randox Laboratories Ltd. Methadone Assay is an in vitro diagnostic test for the qualitative and semi-quantitative detection of Methadone in human urine. The cut off for both the qualitative and semi-quantitative modes of the assay is 300 ng/mL for methadone. The Randox Methadone Assay has been developed for use on the Rx series analyzers, which includes the Rx Daytona and Rx Imola. This in vitro diagnostic device is intended for prescription use only. The semi-quantitative mode is for purposes of 1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GC/MS, or, 2) permitting laboratories to establish quality control procedures. This assay provides only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatograph/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

Device Story

Randox Methadone Assay; in vitro diagnostic enzyme immunoassay; detects methadone in human urine. Input: human urine sample; Reagents: murine monoclonal antibody/substrate reagent and enzyme-drug conjugate reagent. Principle: competitive immunoassay; drug in sample competes with enzyme-labeled drug for antibody binding sites; enzyme activity (G6PDH) inhibited when bound to antibody; free drug allows enzyme activity; NAD converted to NADH; absorbance change measured spectrophotometrically at 340nm. Output: qualitative or semi-quantitative methadone concentration. Used in clinical laboratories; operated by trained personnel on Rx Daytona or Rx Imola analyzers. Output used for preliminary screening; requires GC/MS confirmation for clinical decision-making. Benefits: rapid preliminary identification of methadone presence to guide further confirmatory testing and quality control.

Clinical Evidence

No clinical data provided; bench testing only.

Technological Characteristics

In vitro diagnostic assay; liquid calibrators and controls; utilizes specific Randox analyzers; qualitative and semi-quantitative detection; 300ng/mL cutoff.

Indications for Use

Indicated for the qualitative and semi-quantitative detection of Methadone in human urine for prescription use. Intended for use on specific Randox analyzers to provide preliminary analytical results requiring confirmation by GC/MS.

Regulatory Classification

Identification

A methadone test system is a device intended to measure methadone, an addictive narcotic pain-relieving drug, in serum and urine. Measurements obtained by this device are used in the diagnosis and treatment of methadone use or overdose and to determine compliance with regulations in methadone maintenance treatment.

Special Controls

*Classification.* Class II (special controls). A methadone 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).

Submission Summary (Full Text)

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

A. 510(k) Number:
k092272

B. Purpose for Submission:
New device

C. Measurand:
Methadone

D. Type of Test:
Semi-quantitative and qualitative enzyme immunoassay

E. Applicant:
Randox Laboratories, Limited

F. Proprietary and Established Names:
Randox Methadone

G. Regulatory Information:
Regulation section:

| Regulation | Regulation Name | Class | Product Code | Panel |
| --- | --- | --- | --- | --- |
| 21 CFR§862.3620 | Methadone test system | II | DJR | (91) Toxicology |
| 21 CFR§862.3200 | Clinical toxicology calibrator | II | DKB | (91) Toxicology |
| 21 CFR§862.3280 | Clinical toxicology control material | I, reserved | DIF | (91) Toxicology |

H. Intended Use:

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

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2. Indication(s) for use:

The Randox Laboratories Ltd. Methadone Assay is an in vitro diagnostic test for the qualitative and semi-quantitative detection of Methadone in human urine. The cut off for both the qualitative and semi-quantitative modes of the assay is 300 ng/mL for methadone. The Randox Methadone Assay has been developed for use on the Rx series analyzers, which includes the Rx Daytona and Rx Imola. This in vitro diagnostic device is intended for prescription use only. The semi-quantitative mode is for purposes of

1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GC/MS, or,
2) permitting laboratories to establish quality control procedures.

This assay provides only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatograph/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

Randox Multidrug Calibrator Set

The Randox Multidrug Calibrator Set consists of liquid calibrators containing Oxazepam and Methadone. There are 5 levels of calibrator. They have been developed for use in the calibration of Benzodiazepine (Oxazepam) and Methadone assays on the Rx series analyzers, which includes the Rx Daytona and the Rx Imola. This in vitro diagnostic device is intended for prescription use only.

Randox Multidrug Controls, Level 1 and 2

The Randox Multidrug Controls, level 1 and 2 are liquid controls containing Oxazepam and Methadone. There are 2 levels of controls. They have been developed for use in the quality control of Benzodiazepine and Methadone assays on the Rx series analyzers, which includes the Rx Daytona and the Rx Imola. This in vitro diagnostic device is intended for prescription use only.

3. Special conditions for use statement(s):

Prescription use only

This assay provides only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas chromatograph/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

4. Special instrument requirements:

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The Rx Daytona and Rx Imola analyzers were used to conduct performance studies below.

# I. Device Description:

The Randox Methadone assay is calibrated against methadone. The assay is a two reagent system consisting of a murine monoclonal antibody/substrate reagent and the enzyme-drug conjugate reagent. The Randox Multidrug Calibrator Set consists of 5 calibrators with methadone concentrations from 0 to $1,000\mathrm{ng / mL}$ in human urine. They are ready to use. The Randox Methadone Controls, Level 1 and Level 2, are derived from human urine and are ready to use. The control values are set at approximately $\pm 25\%$ of cutoff $(300\mathrm{ng / mL})$ .

# J. Substantial Equivalence Information:

| 1. Predicate device name(s) | 2. Predicate K number(s) |
| --- | --- |
| DRI® Methadone Assay | k934667 |
| DRI® Multidrug Calibrators and Controls | k983159 |

# 3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device-Randox Methadone Assay | Predicate (k934667) |
| Intended Use/Indications for use | For the qualitative and semi-quantitative determination of Methadone in human urine. | Same |
| Test Principle | A competitive enzyme immunoassay based on competition between drug in the sample and drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. In the absence of drug in the sample the methadone labeled G6PDH conjugate is bound to antibody and enzyme activity is inhibited. When free drug is present in the sample, antibody binds to the free drug and the unbound methadone labeled G6PDH exhibits its maximum enzyme activity. Active | Same |

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| Similarities | | |
| --- | --- | --- |
| Item | Device-Randox Methadone Assay | Predicate (k934667) |
| | enzyme converts NAD to NADH resulting in an absorbance change measured spectrophotometrically at 340nm. | |
| Sample type | Human urine | Same |
| Type of reagent | Liquid Ready to use Two reagent assay | Same |
| Antibody | Monoclonal | Same |
| Cutoff | 300ng/mL | Same |

No significant differences were defined between the predicate (k934667) and new device.

| Similarities | | |
| --- | --- | --- |
| Item | Device-Calibrators and Controls | Predicate (k983159) |
| Intended Use/Indications for use | For the calibration and quality control of the methadone in human urine. | Same |
| Type of reagent | Liquid, ready to use calibrators and controls | Same |
| Controls | 2 levels | Same |
| Differences | | |
| --- | --- | --- |
| Item | Device-Calibrators and Controls | Predicate (k983159) |
| Calibrator materials | Liquid ready to use (0, 150, 300,500, 1000 ng/mL) | Liquid ready to use (150, 300, 500, 1000 ng/mL) |
| Control materials | 225, 375 ng/ml | 200, 375 ng/ml |

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

None were referenced.

# L. Test Principle:

The assay is based on competition between drug in the sample and drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. Enzyme activity decreases upon binding to the antibody, and the drug concentration in the sample is measured in terms of enzyme activity.

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In the absence of drug in the sample, methadone-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. However, when free drug is present in the sample, antibody would bind to free drug; the unbound methadone-labeled G6PDH then exhibits its maximal enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at 340 nm.

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

### 1. Analytical performance:

#### a. Precision/Reproducibility:

A methadone negative urine pool was spiked with methadone to concentrations of ±25%, ±50%, ±75% and ±100% of the cutoff of 300 ng/mL as confirmed by GC/MS. Samples were tested on two RX Daytona and two RX Imola systems twice per day for 20 non-consecutive days (n=80) in the qualitative and semi-quantitative modes. The samples were tested randomly and in a masked order. A summary of the results is below:

RX Daytona

| Sample Concentration % of cutoff | Qualitative Results (#Neg/# Pos) | Semi-Quantitative Results (#Neg/# Pos) |
| --- | --- | --- |
| -100% | 80 Neg | 80 Neg |
| -75% | 80 Neg | 80 Neg |
| -50% | 80 Neg | 80 Neg |
| -25% | 80 Neg | 80 Neg |
| +25% | 80 Pos | 80 Pos |
| +50% | 80 Pos | 80 Pos |
| +75% | 80 Pos | 80 Pos |
| +100% | 80 Pos | 80 Pos |

RX Imola

| Sample Concentration % of cutoff | Qualitative Results (#Neg/# Pos) | Semi-Quantitative Results (#Neg/# Pos) |
| --- | --- | --- |
| -100% | 80 Neg | 80 Neg |
| -75% | 80 Neg | 80 Neg |
| -50% | 80 Neg | 80 Neg |
| -25% | 80 Neg | 80 Neg |
| +25% | 80 Pos | 80 Pos |
| +50% | 80 Pos | 80 Pos |
| +75% | 80 Pos | 80 Pos |
| +100% | 80 Pos | 80 Pos |

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b. Linearity/assay reportable range:

A drug free urine pool was spiked with pure methadone to 1200 ng/mL and serially diluted in increments of 10% with methadone negative urine. Each level was tested in triplicate in the semi-quantitative mode on the Rx Daytona and Rx Imola. % recoveries of the target concentrations were calculated for each instrument. See the tables below:

| | Rx Daytona | | Rx Imola | |
| --- | --- | --- | --- | --- |
| Target (ng/mL) | Result (ng/mL) | % Recovery | Result (ng/mL) | % Recovery |
| 0 | 0.00 | Not applicable | 0.00 | Not applicable |
| 12 | 13.09 | 109.11 | 103.05 | 858.75 |
| 24 | 45.70 | 190.43 | 95.68 | 398.67 |
| 36 | 65.77 | 182.69 | 98.12 | 272.56 |
| 48 | 76.76 | 159.92 | 104.74 | 218.21 |
| 60 | 53.81 | 89.68 | 125.97 | 209.95 |
| 72 | 86.15 | 119.66 | 103.02 | 143.09 |
| 84 | 93.21 | 110.96 | 142.001 | 169.06 |
| 96 | 88.62 | 92.31 | 150.86 | 157.14 |
| 108 | 89.72 | 83.07 | 120.60 | 111.67 |
| 120 | 136.30 | 113.58 | 125.61 | 104.68 |
| 240 | 259.30 | 108.04 | 262.41 | 109.34 |
| 360 | 378.11 | 105.03 | 391.33 | 108.70 |
| 480 | 501.5 | 104.48 | 526.38 | 109.66 |
| 600 | 665.93 | 110.99 | 643.67 | 107.28 |
| 720 | 806.87 | 112.07 | 779.03 | 108.20 |
| 840 | 897.61 | 106.86 | 864.62 | 102.93 |
| 960 | 1013.26 | 105.55 | 978.02 | 101.88 |
| 1080 | 1167.79 | 108.13 | 1070.31 | 99.90 |
| 1200 | 1264.5 | 105.37 | 1118.74 | 93.23 |

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

Traceability and Value Assignment: The Randox Methadone assay has 5 calibrator materials and two control materials that are traceable to master lots that have been GC/MS quantified. The master calibrator and control stock solutions are gravimetrically prepared using methadone from an outside supplier who confirmed the purity of their material by GC/FID, HPLC and NMR. The methadone concentration for the master lot is determined by GC/MS. Value assignment for control materials and calibrator materials is determined against the current master lot by performing nested testing of the new control materials and the new calibrator materials. The new lots are analyzed in replicates of 10. Results need to meet the sponsor's predetermined acceptance criteria %CV and % recovery against the current master lot.

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Calibrator materials and control materials are sold separately.

Stability: Accelerated and Real-time stability studies were performed with the control and calibrator materials at 2-8° C, 25° C and 37° C on the Rx Imola and Rx Daytona. Methadone recoveries from the stressed material were compared to fresh controls or calibrators stored at 2-8° C. Based on the real time studies, the sponsor determined that calibrators and controls are stable for 12 months at 2-8° C. Open vials of calibrators and controls were stored at 2-8° C and analyzed daily on the Rx Imola and Rx Daytona until the methadone recovery exceeded the pre-determined acceptance criteria, which were found to be adequate. Open vials of control materials and calibrator materials are stable for 28 days at 2-8° C.

d. Detection limit:

Performance at low drug concentrations in the semi-quantitative assay was characterized by the determination of recovery (see section b above).

e. Analytical specificity:

The cross-reactivity of the parent drug with endogenous compounds, structurally related, and structurally unrelated compounds was evaluated by adding known amounts of each potential interferent to drug-free urine spiked with methadone to ± 25% of the cutoff (300 ng/mL). Samples were assayed in quintuplets for the qualitative and semi-quantitative modes on the Rx Daytona and Rx Imola for endogenous and structurally unrelated compounds and in triplicate for structurally related compounds. The results were compared to the appropriate sample containing no interferent. Interference was defined as the concentration of each compound that had a response approximately equal to that of the cutoff calibrator (as positive), or the maximum concentration of the compound tested that remained negative. Any compounds demonstrating interference were retested at lower concentrations of the interferent to determine where interference began.

The interfering concentrations of endogenous, structurally unrelated, and structurally related compounds that were equivalent to the cutoff (300 ng/mL) were determined by comparing the absorbance readings of the samples containing the interferent(s) to the absorbance of the calibrator at the cutoff (300 ng/mL). Results are summarized below:

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# Structurally Related Compounds Equivalent to the Cutoff--Rx Daytona

| | | Qualitative | | Semi-Quantitative | | |
| --- | --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Response equivalent to 300 ng/mL cutoff | Cross-reactivity % | Tested Conc. (ng/mL) | Response equivalent to 300 ng/mL cutoff | Cross-reactivity % |
| Methadone | 300 | POS | 100.00% | 300 | POS | 100.00% |
| EDDP | 8592 | POS | 3.49% | 5588 | POS | 5.37% |
| EMDP | 805634 | POS | 0.04% | 342375 | POS | 0.09% |
| (-) alpha Methadol | 232 | POS | 129.17% | 205 | POS | 146.22% |
| LAAM | 310 | POS | 96.86% | 281 | POS | 106.94% |
| d, l-Alpha methadol | 304 | POS | 98.70% | 303 | POS | 98.94% |

# Structurally Related Compounds Equivalent to the Cutoff-- Rx Imola

| | | Qualitative | | Semi-Quantitative | | |
| --- | --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Response equivalent to 300 ng/mL cutoff | Cross-reactivity % | Tested Conc. (ng/mL) | Response equivalent to 300 ng/mL cutoff | Cross-reactivity % |
| Methadone | 300 | POS | 100.00% | 300 | POS | 100.00% |
| EDDP | 7389 | POS | 4.06% | 5882 | POS | 5.10% |
| EMDP | 713993 | POS | 0.04% | 446281 | POS | 0.07% |
| (-) alpha Methadol | 225 | POS | 133.46% | 205 | POS | 146.10% |
| LAAM | 294 | POS | 101.94% | 300 | POS | 100.12% |
| d, l-Alpha methadol | 293 | POS | 102.42% | 289 | POS | 103.98% |

Structurally unrelated compounds were tested by spiking each of them into urine samples that contain methadone at $\pm 25\%$ of cutoff for the Rx Imola and Rx Daytona. The following is the list of compounds tested; 11-hydroxy-delta9-THC, 11-nor9-carboxy-delta9-THC, 6 Acetyl-morphine, Amitriptyline, Amobarbital, $(+/-)$-Amphetamine, Aspirin, Ascorbic acid, Benzoylecgonine, $\beta$-phenylethylamine, Buprenorphine, Caffeine, Cannabidiol, Chlorpheniramine, Cocaethylene, Cocaine, Codeine, Cotinine, delta9-THC, Diazepam, Dihydrocodeine, Ecgonine methyl ester, D,l-Ephedrine, l-Ephedrine, d-Ephedrine, S,S (+) Pseudoephedrine, R,R (-) Pseudoephedrine, Heroin, MBDB, MDA, MDEA, MDMA, d,l-Methamphetamine, Morphine, Oxycodone, Paracetamol, Temazepam, Ibuprofen. Results are summarized in the labeling. Interference at the cutoff is summarized below:

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Structurally Unrelated Compounds Equivalent to the Cutoff--Rx Daytona

| | | Response 300 ng/mL | equivalent to cutoff | Cross- reactivity % | |
| --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Qualitative | Semi-quantitative | Qualitative | Semi-quantitative |
| 11-hydroxy-delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| 11-nor9-carboxy-delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| 6 Acetyl-morphine | 100,000 | NEG | NEG | 0% | 0% |
| Amitriptyline | 100,000 | NEG | NEG | 0.27% | 0.35% |
| Amobarbital | 100,000 | NEG | NEG | 0% | 0% |
| | | | | | |
| (+/-)-Amphetamine | 100,000 | NEG | NEG | 0% | 0% |
| Aspirin | 100,000 | NEG | NEG | 0% | 0% |
| Ascorbic acid | 100,000 | NEG | NEG | 0% | 0% |
| Benzoylecgonine | 100,000 | NEG | NEG | 0% | 0% |
| β-phenylethylamine | 100,000 | NEG | NEG | 0% | 0% |
| Buprenorphine | 100,000 | NEG | NEG | 0% | 0% |
| Caffeine | 100,000 | NEG | NEG | 0% | 0% |
| Cannabidiol | 100,000 | NEG | NEG | 0% | 0% |
| Chlorpheniramine | 4,000 | NEG | NEG | 5.00% | 5.25% |
| Cocaethylene | 100,000 | NEG | NEG | 0% | 0% |
| Cocaine | 100,000 | NEG | NEG | 0% | 0% |
| Codeine | 100,000 | NEG | NEG | 0% | 0% |
| Cotinine | 100,000 | NEG | NEG | 0% | 0% |
| delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| Diazepam | 100,000 | NEG | NEG | 0% | 0% |
| Dihydrocodeine | 100,000 | NEG | NEG | 0% | 0% |
| Ecgonine methyl ester | 100,000 | NEG | NEG | 0% | 0% |
| D,l-Ephedrine | 100,000 | NEG | NEG | 0% | 0% |
| l-Ephedrine | 100,000 | NEG | NEG | 0% | 0% |
| d-Ephedrine | 100,000 | NEG | NEG | 0% | 0% |
| S,S (+) Pseudoephedrine | 100,000 | NEG | NEG | 0.02% | 0.06% |
| R,R (-) Pseudoephedrine | 100,000 | NEG | NEG | 0.02% | 0.05% |
| Heroin | 100,000 | NEG | NEG | 0% | 0% |
| MBDB | 100,000 | NEG | NEG | 0.10% | 0.07% |
| MDA | 100,000 | NEG | NEG | 0% | 0% |
| MDEA | 100,000 | NEG | NEG | 0.03% | 0.05% |
| MDMA | 100,000 | NEG | NEG | 0.03% | 0.07% |
| d,l-Methamphetamine | 100,000 | NEG | NEG | 0.05% | 0.08% |
| Morphine | 100,000 | NEG | NEG | 0% | 0% |
| Oxycodone | 100,000 | NEG | NEG | 0% | 0% |
| Paracetamol | 100,000 | NEG | NEG | 0% | 0% |
| Temazepam | 100,000 | NEG | NEG | 0% | 0% |
| Ibuprofen | 100,000 | NEG | NEG | 0% | 0% |

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Structurally Unrelated Compounds Equivalent to the Cutoff--Rx Imola

| | | Response 300 ng/mL | equivalent to cutoff | Cross- | reactivity % |
| --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Qualitative | Semi-quantitative | Qualitative | Semi-quantitative |
| 11-hydroxy-delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| 11-nor9-carboxy-delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| 6 Acetyl-morphine | 100,000 | NEG | NEG | 0% | 0% |
| Amitriptyline | 60,000 | NEG | NEG | 0.39% | 0.34% |
| Amobarbital | 100,000 | NEG | NEG | 0% | 0% |
| (+/-)-Amphetamine | 100,000 | NEG | NEG | 0% | 0% |
| Aspirin | 100,000 | NEG | NEG | 0% | 0% |
| Ascorbic acid | 100,000 | NEG | NEG | 0% | 0% |
| Benzoylecgonine | 100,000 | NEG | NEG | 0% | 0% |
| β-phenylethylamine | 100,000 | NEG | NEG | 0% | 0% |
| Buprenorphine | 100,000 | NEG | NEG | 0% | 0% |
| Caffeine | 100,000 | NEG | NEG | 0% | 0% |
| Cannabidiol | 100,000 | NEG | NEG | 0% | 0% |
| Chlorpheniramine | 5,000 | NEG | NEG | 5.67% | 5.29% |
| Cocaethylene | 100,000 | NEG | NEG | 0% | 0% |
| Cocaine | 100,000 | NEG | NEG | 0% | 0% |
| Codeine | 100,000 | NEG | NEG | 0% | 0% |
| Cotinine | 100,000 | NEG | NEG | 0% | 0% |
| delta9-THC | 100,000 | NEG | NEG | 0% | 0% |
| Diazepam | 100,000 | NEG | NEG | 0% | 0% |
| Dihydrocodeine | 100,000 | NEG | NEG | 0% | 0% |
| Ecgonine methyl ester | 100,000 | NEG | NEG | 0% | 0% |
| d,l-Ephedrine | 100,000 | NEG | NEG | 0% | 0.03% |
| l-Ephedrine | 100,000 | NEG | NEG | 0% | 0% |
| d-Ephedrine | 100,000 | NEG | NEG | 0% | 0% |
| S,S (+) Pseudoephedrine | 100,000 | NEG | NEG | 0.02% | 0.05% |
| R,R (-) Pseudoephedrine | 100,000 | NEG | NEG | 0.02% | 0.06% |
| Heroin | 100,000 | NEG | NEG | 0% | 0% |
| MBDB | 100,000 | NEG | NEG | 0.06% | 0.27% |
| MDA | 100,000 | NEG | NEG | 0% | 0% |
| MDEA | 100,000 | NEG | NEG | 0.03% | 0.06% |
| MDMA | 100,000 | NEG | NEG | 0.04% | 0.08% |
| d,l-Methamphetamine | 100,000 | NEG | NEG | 0% | 0% |
| Morphine | 100,000 | NEG | NEG | 0% | 0% |
| Oxycodone | 100,000 | NEG | NEG | 0% | 0% |
| Paracetamol | 100,000 | NEG | NEG | 0% | 0% |
| Temazepam | 100,000 | NEG | NEG | 0.02% | 0.06% |
| Ibuprofen | 100,000 | NEG | NEG | 0% | 0% |

Endogenous interfering compounds were tested at $\pm 25\%$ of cutoff for the Rx Imola and Rx Daytona for Total Bilirubin, Direct Bilirubin, Hemoglobin,

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Creatinine, Urea, Glucose, H.S.A., Ethanol, Acetone, Gamma globulin, Oxalic acid, Riboflavin, Sodium chloride, Boric acid, Sodium azide, Sodium fluoride. Results are summarized in the labeling. Interference at the cutoff is in the table below. No interference was observed at the cutoff.

## Endogenous Interference Equivalent to the Cutoff—Rx Daytona

| | | Response 300 ng/mL | equivalent to cutoff | Cross- reactivity % | |
| --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Qualitative | Semi-quantitative | Qualitative | Semi-quantitative |
| Total Bilirubin | 15 | NEG | NEG | 0% | 0% |
| Direct Bilirubin | 5 | NEG | NEG | 0% | 0% |
| Haemoglobin | 115 | NEG | NEG | 0% | 0% |
| Creatinine | 30 | NEG | NEG | 0% | 0% |
| Urea | 258 | NEG | NEG | 0% | 0% |
| Glucose | 2000 | NEG | NEG | 0% | 0% |
| H.S.A. | 500 | NEG | NEG | 0% | 0% |
| Ethanol | 1000 | NEG | NEG | 0% | 0% |
| Acetone | 1000 | NEG | NEG | 0% | 0% |
| Gamma globulin | 500 | NEG | NEG | 0% | 0% |
| Oxalic acid | 100 | NEG | NEG | 0% | 0% |
| Riboflavin | 7.5 | NEG | NEG | 0% | 0% |
| Sodium chloride | 6000 | NEG | NEG | 0% | 0% |
| Boric acid | 1000 | NEG | NEG | 0% | 0% |
| Sodium azide | 1000 | NEG | NEG | 0% | 0% |
| Sodium fluoride | 1000 | NEG | NEG | 0% | 0% |

## Endogenous Interference Equivalent to the Cutoff—Rx Imola

| | | Response 300 ng/mL | equivalent to cutoff | Cross- reactivity % | |
| --- | --- | --- | --- | --- | --- |
| Compound | Tested Conc. (ng/mL) | Qualitative | Semi-quantitative | Qualitative | Semi-quantitative |
| Total Bilirubin | 15 | NEG | NEG | 0% | 0% |
| Direct Bilirubin | 5 | NEG | NEG | 0% | 0% |
| Haemoglobin | 115 | NEG | NEG | 0% | 0% |
| Creatinine | 30 | NEG | NEG | 0% | 0% |
| Urea | 258 | NEG | NEG | 0% | 0% |
| Glucose | 2000 | NEG | NEG | 0% | 0% |
| H.S.A. | 500 | NEG | NEG | 0% | 0% |
| Ethanol | 1000 | NEG | NEG | 0% | 0% |
| Acetone | 1000 | NEG | NEG | 0% | 0% |
| Gamma globulin | 500 | NEG | NEG | 0% | 0% |
| Oxalic acid | 100 | NEG | NEG | 0% | 0% |
| Riboflavin | 7.5 | NEG | NEG | 0% | 0% |
| Sodium chloride | 6000 | NEG | NEG | 0% | 0% |
| Boric acid | 1000 | NEG | NEG | 0% | 0% |
| Sodium azide | 1000 | NEG | NEG | 0% | 0% |
| Sodium fluoride | 1000 | NEG | NEG | 0% | 0% |

Specific Gravity studies were performed on the Rx Daytona and Rx Imola. Two methadone negative urine pools were spiked with methadone to + and – 25% cutoff (300 ng/mL). The pools were further divided and had sodium

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chloride added to achieve specific gravities of 1.000 (water blank), 1.010, 1.020, 1.030. Specific gravities were confirmed by a commercial method. Each sample was analyzed in replicates of 10 in the semi-quantitative and qualitative modes. Recoveries were compared to the spiked target samples and were expected to be $\leq 10\%$. Specific gravity from 1.000-1.030 had no significant ($\leq 10\%$) effect on EDDP results for the Rx Daytona or Rx Imola.

pH studies were performed by spiking methadone into drug free negative urine to achieve methadone concentrations at $\pm 25\%$ cutoff. pH was adjusted by adding either 1M HCl or 1M NaOH to obtain samples with pH levels of 3, 5, 7, 9, 11. pH was confirmed by a pH meter. Each sample was analyzed in replicates of 10 in the semi-quantitative and qualitative modes. Recoveries were compared to the pH 7 sample and were expected to be $\leq 10\%$ of the target values. pH from 3-11 had no significant ($\leq 10\%$) effect on methadone results for the Rx Daytona or Rx Imola.

f. Assay cut-off:

Analytical performance of the device around the claimed cutoff is described in precision section (1 a.) above.

2. Comparison studies:

a. Method comparison with predicate device:

54 positive and 45 negative unaltered natural samples were analyzed on the Rx Daytona and Rx Imola and by GC/MS. The results with discordant tables are as follows:

RX Daytona Semi-quantitative

| Candidate Device Results | Negative | Less than half the cut-off (concentration by GC/MS analysis) | Near Cutoff Negative (Between 50% below the cut-off and the cut-off concentration) | Near Cutoff Positive (Between the cut-off and 50% above the cut-off concentration) | High Positive (greater than 50% above the cut-off concentration) | % Agreement with GC/MS |
| --- | --- | --- | --- | --- | --- | --- |
| Positive | - | - | 9 | 10 | 35 | 100.00% |
| Negative | 39 | - | 6 | - | - | 83.33% |
| Cutoff Value | Semi-quantitative Methadone Rx Daytona | Drug/Metabolite GC/MS Methadone (ng/mL) |
| --- | --- | --- |
| 300 ng/mL | Pos | 240 |
| | Pos | 260 |
| | Pos | 280 |
| | Pos | 290 |
| | Pos | 290 |
| | Pos | 295 |
| | Pos | 298 |
| | Pos | 298 |
| | Pos | 298 |

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RX Daytona Qualitative

| Candidate Device Results | Negative | Less than half the cut-off (concentration by GC/MS analysis) | Near Cutoff Negative (Between 50% below the cut-off and the cut-off concentration) | Near Cutoff Positive (Between the cut-off and 50% above the cut-off concentration) | High Positive (greater than 50% above the cut-off concentration) | % Agreement with GC/MS |
| --- | --- | --- | --- | --- | --- | --- |
| Positive | - | - | 8 | 10 | 35 | 100.00% |
| Negative | 39 | - | 7 | - | - | 85.19% |
| Cutoff Value | Qualitative Methadone Rx Daytona | Drug/Metabolite GC/MS Methadone (ng/mL) |
| --- | --- | --- |
| 300 ng/mL | Pos | 240 |
| | Pos | 260 |
| | Pos | 280 |
| | Pos | 290 |
| | Pos | 290 |
| | Pos | 295 |
| | Pos | 298 |
| | Pos | 298 |

RX Imola Semi-quantitative

| Candidate Device Results | Negative | Less than half the cut-off (concentration by GC/MS analysis) | Near Cutoff Negative (Between 50% below the cut-off and the cut-off concentration) | Near Cutoff Positive (Between the cut-off and 50% above the cut-off concentration) | High Positive (greater than 50% above the cut-off concentration) | % Agreement with GC/MS |
| --- | --- | --- | --- | --- | --- | --- |
| Positive | - | - | 7 | 10 | 35 | 100.00% |
| Negative | 39 | - | 8 | - | - | 87.03% |
| Cutoff Value | Qualitative Methadone Rx Imola | Drug/Metabolite GC/MS Methadone (ng/mL) |
| --- | --- | --- |
| 300 ng/mL | Pos | 240 |
| | Pos | 260 |
| | Pos | 280 |
| | Pos | 290 |
| | Pos | 295 |
| | Pos | 298 |
| | Pos | 298 |

RX Imola Qualitative

| Candidate Device Results | Negative | Less than half the cut-off (concentration by GC/MS analysis) | Near Cutoff Negative (Between 50% below the cut-off and the cut-off concentration) | Near Cutoff Positive (Between the cut-off and 50% above the cut-off concentration) | High Positive (greater than 50% above the cut-off concentration) | % Agreement with GC/MS |
| --- | --- | --- | --- | --- | --- | --- |
| Positive | - | - | 9 | 10 | 35 | 100.00% |
| Negative | 39 | - | 6 | - | - | 83.33% |

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| Cutoff Value | Qualitative Methadone Rx Imola | Drug/Metabolite GC/MS Methadone (ng/mL) |
| --- | --- | --- |
| 300 ng/mL | Pos | 240 |
| | Pos | 260 |
| | Pos | 280 |
| | Pos | 290 |
| | Pos | 290 |
| | Pos | 295 |
| | Pos | 298 |
| | Pos | 298 |
| | Pos | 298 |

b. Matrix comparison:

Not applicable. This device is for urine only.

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):

Not applicable

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.