EMIT II PLUS ECSTASY ASSAY

K043028 · Dade Behring, Inc. · DKZ · Jan 7, 2005 · Clinical Toxicology

Device Facts

• Record ID: K043028
• Device Name: EMIT II PLUS ECSTASY ASSAY
• Applicant: Dade Behring, Inc.
• Product Code: DKZ · Clinical Toxicology
• Decision Date: Jan 7, 2005
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3100
• Device Class: Class 2

Indications for Use

The Emit® II Plus Ecstasy Assay is a homogeneous enzyme immunoassay with a 300 ng/mL or 500 ng/mL cutoff. The assay is intended for use in laboratories for the qualitative and/or semiquantitative analysis of methylenedioxymethamphetamine (MDMA) and closely related drugs in human urine. Emit® II Plus Assays are designed for use with a number of chemistry analyzers. The Emit® II Plus Ecstasy Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Other chemical confirmation methods are available. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when preliminary positive results are used. The Emit® II Plus Ecstasy Calibrators / Controls are used in the calibration of the Emit® II Plus Ecstasy Assay. These standards may also be used as quality control materials based upon the Ecstasy assay cutoff.

Device Story

Homogeneous enzyme immunoassay for MDMA detection in human urine; utilizes automated clinical chemistry analyzers. Input: urine sample; Reagent 1 (sheep polyclonal antibodies to MDMA, G6P, NAD); Reagent 2 (MDA labeled with bacterial recombinant G6PDH). Principle: competitive binding between drug in urine and drug-labeled enzyme for antibody sites; enzyme activity inversely proportional to drug concentration. Output: absorbance change measured spectrophotometrically at 340 nm; qualitative or semi-quantitative drug concentration. Used in clinical laboratories by technicians; results are preliminary and require GC/MS confirmation. Benefits: rapid screening for MDMA and related compounds to aid clinical decision-making.

Clinical Evidence

Bench testing only. Performance evaluated using 100 urine specimens compared to GC/MS reference method. At 300 ng/mL cutoff: 98% agreement (55/57 positive, 43/43 negative). At 500 ng/mL cutoff: 96% agreement (53/57 positive, 43/43 negative). Discrepant samples were within ±50% of the cutoff.

Technological Characteristics

Homogeneous enzyme immunoassay; sheep polyclonal antibodies to MDMA; bacterial G6PDH enzyme label; spectrophotometric detection of NADH production; liquid form; compatible with chemistry analyzers; includes preservatives and stabilizers (bovine serum albumin).

Indications for Use

Indicated for laboratory use for qualitative and/or semiquantitative analysis of MDMA and related drugs in human urine. Not for point-of-care settings.

Regulatory Classification

Identification

An amphetamine test system is a device intended to measure amphetamine, a central nervous system stimulating drug, in plasma and urine. Measurements obtained by this device are used in the diagnosis and treatment of amphetamine use or overdose and in monitoring levels of amphetamine to ensure appropriate therapy.

Special Controls

*Classification.* Class II (special controls). An amphetamine 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

• Microgenics DRI® Ecstasy Enzyme Immunoassay (k012110)

Submission Summary (Full Text)

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

A. 510(k) Number:
k043028

B. Purpose for Submission:
New Device

C. Measurand:
Methylenedioxymethamphetamine (MDMA)

D. Type of Test:
Qualitative and semi-quantitative immunoassay

E. Applicant:
Dade Behring, Inc.

F. Proprietary and Established Names:
Dade Behring Syva® Emit® II Plus Ecstasy Assay

G. Regulatory Information:
1. Regulation section:
21 CFR 862.3100 (Enzyme Immunoassay, Amphetamine)
21 CFR 862.3200 (Calibrators, Drug Specific)
21 CFR 862.3280 (Clinical Toxicology Control Material)
2. Classification:
Class II
Class II
Class I
3. Product Code:
DKZ
DLJ
DIF
4. Panel:
Toxicology (91)

H. Intended Use:
1. Intended use(s):
Refer to Indications for use.
2. Indication(s) for use:
The Emit® II Plus Ecstasy Assay is a homogeneous enzyme immunoassay with a 300 ng/mL or 500 ng/mL cutoff. The assay is intended for use in

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laboratories for the qualitative and/or semiquantitative analysis of methylenedioxymethamphetamine (MDMA) and closely related drugs in human urine. Emit® II Plus Assays are designed for use with a number of chemistry analyzers.

The Emit® II Plus Ecstasy Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Other chemical confirmation methods are available. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

The Emit® II Plus Ecstasy Calibrators/Controls are used in the calibration of the Emit® II Plus Ecstasy Assay. These standards may also be used as quality control materials based upon the Ecstasy assay cutoff.

3. Special condition for use statement(s):
Semi-quantitative results may be helpful in estimating the concentrations of drug(s) in samples. This can aid users when they are preparing dilutions of the samples for further analysis.

The assay is not designated for use in point-of-care settings.

Certain foods or medications may interfere with tests for methylenedioxymethamphetamine and cause false positive results.

4. Special instrument Requirements:
The device is for use on automated clinical chemistry analyzers. Instruments must be capable of maintaining a constant reaction temperature, pipetting samples and reagents, mixing reagents, timing reactions, and measuring enzyme rates precisely.

Performance was demonstrated in this submission on the SYVA®-30R Biochemical System.

I. Device Description:
The device consists of two wet reagents which contain the key components of the immunoassay; Antibody/Substrate Reagent 1 contains sheep polyclonal antibodies to MDMA, bovine serum albumin, glucose-6-phosphate, nicotinamide adenine dinucleotide, preservatives, and stabilizers. Enzyme Reagent 2 contains methylenedioxyamphetamine (MDA) labeled with bacterial recombinant glucose-6-phosphate dehydrogenase, Tris buffer, bovine serum albumin, preservatives, and stabilizers.

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Five levels of calibrator/control (Level 0 - Level 4) are provided separately:

| Desired Cutoff Level (ng/mL) | Additional Recommended Calibrators/Controls for Qualitative Analysis (ng/mL) | Required Calibrators/Controls for Semiquantitative Analysis (ng/mL) |
| --- | --- | --- |
| 300 (Level 2) | *Level 0 (0) | Level 0 (0) |
| | Level 4 (1000) | Level 1 (150) |
| 500 (Level 3) | Level 0 (0) | Level 2 (300) |
| | Level 4 (1000) | Level 3 (500) |
| | | Level 4 (1000) |

For any individual cutoff level, a calibrator/control is used as either a calibrator or as a control when the assay is used for qualitative analysis. When a calibrator/control is used as a calibrator for an individual cutoff level, the other level calibrators/controls (above or below it, as listed above) are used as controls.

*Emit® Calibrator/Control Level 0 was previously cleared under k993755

# J. Substantial Equivalence Information:

1. Predicate device name(s):

Microgenics DRI® Ecstasy Enzyme Immunoassay

2. Predicate K number(s):

k012110

3. Comparison with predicate:

Both devices are for measurement of the same analyte in the same matrix and utilize the same test methodology. The predicate device has a single cutoff of $500\mathrm{ng / mL}$ . The user can choose a cutoff of 300 or $500\mathrm{ng / mL}$ with the candidate device. Both are for use on automated analyzers.

The reagent formulations vary between the two devices.

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Intended Use | Same | Qualitative or semi-quantitative determination of MDMA and related drugs in human urine |
| Principle | Same | Homogeneous enzyme immunoassay |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Cutoff | 300 or 500 ng/mL | 500 ng/mL |
| Antibody | Polyclonal | Monoclonal |
| Components of | Sheep polyclonal antibodies to | Monoclonal anti-MDMA |
| antigen | Serum IgG and IgM antibodies to | Monoclonal anti-MDMA |
| antigen | Serum IgG and IgM antibodies to | Monoclonal anti-MDMA |

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| Antibody/Substrate Reagent | methylenedioxymethamphetamine (MDMA), bovine serum albumin, G6P, NAD, preservatives and stabilizers. | antibody, G6P, NAD in tris buffer with sodium azide as a preservative. |
| --- | --- | --- |
| Components of Enzyme Reagent | Methylenedioxyamphetamine (MDA) labeled with bacterial G6PDH, tris buffer, bovine serum albumin, preservatives and stabilizers. | Methylenedioxymethamphetamine (MDMA) labeled with G6PDH, tris buffer, and sodium azide as a preservative. |

K. Standard/Guidance Document Referenced (if applicable):
The sponsor referenced the following guidance document or standards in their submission:

Premarket Submission and Labeling Recommendations for Drugs of Abuse Screening Tests - Draft Guidance for Industry and FDA Staff, published December 2003

NCCLS EP5-A: Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline

L. Test Principle:
The test is an enzyme immunoassay for use on automated clinical chemistry analyzers. For qualitative results, two calibrators at 0 and 1000 ng/mL are run with the assay. For semi-quantitative results five calibrators ranging in concentration from 0 to 1000 ng/mL are run with the assay. The assay is based upon competition between drug in the urine specimen and drug labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, so the drug concentration in the specimen can be measured in terms of enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous serum G6PDH does not interfere because the coenzyme NAD functions only with the bacterial (Leuconstoc mesenteroides) enzyme employed in the assay.

Absorbance measurements are taken at 340 nm. Control and unknown values are read from the calibration curve.

M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
All performance was established on the SYVA®-30R Biochemical System.
Specimen description: calibrator/control material consisting of human urine, MDMA, and preservatives.
Number of days: 20

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Replicates per day: 4

Lots of product used: 1

Number of calibrations performed during the study: 1

Results of the studies are presented below.

Qualitative Precision

| Sample concentration, ng/mL | Mean mA/min | SD | CV% | Sample concentration, ng/mL | Mean mA/min | SD | CV% |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Within-Run Imprecision | | | | Total Imprecision | | | |
| 225 | 383 | 1.33 | 0.35 | 225 | 383 | 2.11 | 0.55 |
| 300 | 419 | 2.33 | 0.56 | 300 | 419 | 3.14 | 0.75 |
| 375 | 455 | 2.03 | 0.45 | 375 | 455 | 2.81 | 0.62 |
| 500 | 495 | 3.91 | 0.79 | 500 | 495 | 4.21 | 0.85 |
| 625 | 522 | 2.13 | 0.41 | 625 | 522 | 4.97 | 0.95 |

Semi-quantitative Precision

| Sample concentration, ng/mL | Mean ng/mL | SD | CV% | Sample concentration, ng/mL | Mean ng/mL | SD | CV% |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Within-Run Imprecision | | | | Total Imprecision | | | |
| 225 | 220 | 2.56 | 1.16 | 225 | 220 | 4.06 | 1.85 |
| 300 | 291 | 4.93 | 1.69 | 300 | 291 | 6.63 | 2.28 |
| 375 | 375 | 5.28 | 1.41 | 375 | 375 | 7.28 | 1.94 |
| 500 | 502 | 11.06 | 2.20 | 500 | 502 | 14.07 | 2.80 |
| 625 | 638 | 12.39 | 1.94 | 625 | 638 | 17.09 | 2.68 |

b. Linearity/assay reportable range:

Support for the lower limit of the reportable range is characterized by the sensitivity studies and is supported at the high end by the highest calibrator used by the assay.

Recovery studies

Sample description: Negative Human Urine

Number of replicates: 5

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| Expected Concentration | Mean Observed Concentration | % Recovery | Mean Absorbance Reading | Standard Deviation | Coefficient of Variation (%) |
| --- | --- | --- | --- | --- | --- |
| 150 | 144.4 | 96.3 | 353.8 | 1.6 | 0.5 |
| 200 | 181.5 | 90.8 | 372.1 | 2.1 | 0.6 |
| 225 | 204.4 | 90.8 | 383.7 | 1.7 | 0.4 |
| 250 | 230.2 | 92.1 | 396.7 | 1.2 | 0.3 |
| 270 | 249.1 | 92.2 | 406.3 | 1.1 | 0.3 |
| 300 | 274.5 | 91.5 | 418.8 | 1.3 | 0.3 |
| 330 | 315.2 | 95.5 | 437.8 | 2.1 | 0.5 |
| 375 | 356.8 | 95.1 | 455.4 | 1.4 | 0.3 |
| 450 | 434.2 | 96.5 | 483.0 | 2.1 | 0.4 |
| 500 | 492.3 | 98.5 | 501.0 | 1.4 | 0.3 |
| 550 | 542.4 | 98.6 | 513.7 | 1.6 | 0.3 |
| 625 | 631.0 | 101.0 | 531.9 | 0.7 | 0.1 |
| 750 | 740.0 | 98.7 | 548.6 | 0.9 | 0.2 |
| 900 | 849.0 | 94.3 | 560.8 | 1.3 | 0.2 |
| 1000 | 891.7 | 89.2 | 564.6 | 2.2 | 0.4 |

# c. Traceability (controls, calibrators, or method):

Five levels of calibrator/control material, ranging in concentration from 0 to $1000\mathrm{ng / mL}$ , are specified in the labeling but are supplied separately. Calibrators/controls are drug free urine based materials spiked with known concentrations of MDMA.

After a master lot of each level of calibrator/control is prepared gravimetrically, the assigned values of the master lot are verified by GC/MS analysis. GC/MS concentrations must be within $\pm 10\%$ of the target MDMA concentration. Each new calibrator/control lot is prepared in the same manner as the master lot and run as an unknown vs. the master lot. The concentration of the new calibrator lot is adjusted, if needed, to be within $\pm 5\%$ of the mean value obtained for the corresponding level of the master lot. The value of the new lot is also confirmed by GC/MS.

Stability studies are summarized for the calibrators/controls.

The shelf life of the calibrators/controls will be established based on real-time data from 3 lots. Calibrators/controls stored at $2 - 8^{\circ}\mathrm{C}$ are compared to a standard curve generated from calibrators stored at $-20^{\circ}\mathrm{C}$ . The time points tested will be day 0, 7, 14, 21, and 30, and monthly thereafter up to 13 months. Nine replicates will be averaged at each time point. The observed drift of the mean concentration must be within $\pm 10\%$ of the reference calibrators. Shelf life stability claims at the time of marketing will be based on

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real-time data, which is expected to be 8 months. Shelf life will be extended as additional real-time data becomes available.

Opened bottle stability testing will be performed on all levels. At day 0, portions of the bottle will be removed and analyzed. The bottles are then recapped and stored upright at 2-8°C. The process is repeated at 7 months and 13 months. Twenty-five or greater replicates will be averaged at each time point. At each time point, the observed drift of the mean concentration must be within ± 10% of the concentration at day 0. The sponsor states that for opened bottle stability testing, the number of replicate measurements is larger than for closed bottle stability to provide statistically valid results using fewer time points.

d. Detection limit:
Sensitivity of the assay is less than 75 ng/mL. To determine analytical sensitivity, the sponsor assayed the negative calibrator 20 times within the same run and extrapolated the value of each measurement from the standard curve. The average and standard deviation of those readings were calculated. The analytical sensitivity was estimated by adding 2 standard deviations to the average of the readings.

e. Analytical specificity:
Cross-reactivity was established by spiking various concentrations of similarly structured drug compounds into drug-free urine. By analyzing various concentration of each compound the sponsor determined the concentration of the drug that produced a response approximately equivalent to the cutoff concentration of the assay.

| Drug Compound | Concentration in ng/mL producing response equivalent to the 300 cutoff | Response equivalent to the 500 cutoff (ng/mL) |
| --- | --- | --- |
| d-amphetamine | >49,000 | >265,000 |
| l-amphetamine | >63,000 | >300,000 |
| d-methamphetamine | >12,000 | >57,000 |
| l-methamphetamine | >9,000 | >13,000 |
| 3,4-Methylenedioxyethylamphetamine (MDEA) | 286 | 528 |
| N-Methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine hydrochloride (MBDB) | 168 | 365 |
| 3,4-Methylenedioxyamphetamine (MDA) | 284 | 578 |
| 3,4-Methylenedioxyphenyl-2-butanamine hydrochloride (BDB) | 152 | 347 |
| p-Methoxyamphetamine hydrochloride (PMA) | 4,933 | 22,024 |
| p-Methoxymethamphetamine hydrochloride (PMMA) | 1,433 | 3,387 |

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| 4-Hydroxy-3-methoxymethamphetamine (HMMA) | >40,000 | >40,000 |
| --- | --- | --- |
| d,l-amphetamine | >54,000 | >300,000 |
| d,l-methamphetamine | >9,000 | >51,000 |
| 4-Chloramphetamine | >2,000 | >6,000 |
| Benzphetamine | >10,000 | >10,000 |
| Buproprion | >500,000 | >500,000 |
| Chloroquine | >1,000,000 | >1,000,000 |
| l-Ephedrine | >52,000 | >384,000 |
| Fenfluramine | >1,000 | >6,000 |
| Mephentermine | >15,000 | >15,000 |
| Methoxyphenamine | >1,000,000 | >1,000,000 |
| Nor-pseudoephedrine | >100,000 | >100,000 |
| Phenmetrazine | >300,000 | >300,000 |
| Phentermine | >150,000 | >150,000 |
| Phenylpropanolamine | >200,000 | >200,000 |
| Propanolol | >15,000 | >15,000 |
| Pseudoephedrine | >58,000 | >411,000 |
| Quinacrine | >1,000,000 | >1,000,000 |
| Tranylcypromine | >100,000 | >100,000 |
| Tyramine | >100,000 | >100,000 |
| Haloperidol | >1,000 | >2,500 |
| Isoxsuprine | >2,500 | >10,000 |
| Ketorolac Tromethamine | >50,000 | >100,000 |
| Labelatol | >10,000 | >25,000 |
| Nylidrin | >5,000 | >7,500 |
| Trazadone | >1,000 | >2,500 |

The following compounds were evaluated for potential positive interference with the assay. To evaluate for interference the sponsor spiked potentially interfering compounds into drug-free urine. The sponsor recorded the greatest concentration of potential interferent tested that produced a negative result. That concentration appears in the table. Results of the study are presented below:

| Potential Interferent | Concentration in μg/mL producing negative response at the 300 cutoff | Concentration in μg/mL producing negative response at the 500 cutoff |
| --- | --- | --- |
| Acetaminophen | 1000 | 1000 |
| α-Acetyl-N, N-dinormethadol | 25 | 25 |
| 1-α-Acetylmethadol (LAAM) | 25 | 25 |
| N-Acetylprocainamide (NAPA) | 400 | 400 |

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| Acetylsalicylic Acid | 1000 | 1000 |
| --- | --- | --- |
| Albuterol | 1000 | 1000 |
| p-Aminobenzoic Acid (PABA) | 1000 | 1000 |
| Amitriptyline | 100 | 500 |
| Amoxicillin | 100 | 100 |
| Atenolol | 1000 | 1000 |
| Benzoylecgonine | 1000 | 1000 |
| Buprenorphine | 100 | 100 |
| Caffeine | 1000 | 1000 |
| Carbamazepine | 250 | 250 |
| Carisoprodol | 1000 | 1000 |
| Chlorpheniramine | 100 | 100 |
| Chlorpromazine | 200 | 200 |
| Cimetidine | 1000 | 1000 |
| Clomipramine | 2.5 | 2.5 |
| Clonidine | 1000 | 1000 |
| Codeine | 500 | 500 |
| Cotinine | 100 | 100 |
| Cyclobenzaprine | 28 | 28 |
| Desipramine | 800 | 800 |
| Dextromethorphan | 1000 | 1000 |
| Dextrorphan | 280 | 280 |
| Diphenhydramine | 1000 | 1000 |
| Doxepin | 10 | 10 |
| Doxylamine | 500 | 500 |
| Epinephrine | 1000 | 1000 |
| 2-Ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) | 1000 | 1000 |
| Fenoprofen | 1000 | 1000 |
| Fluoxetine | 100 | 500 |
| Furosemide | 1000 | 1000 |
| Glutethimide | 500 | 500 |
| Ibuprofen | 1000 | 1000 |
| Imipramine | 100 | 400 |
| Ketamine | 100 | 100 |
| Ketoprofen | 1000 | 1000 |
| Lidocaine | 100 | 100 |
| LSD | 0.15 | 0.15 |
| Meperidine HCl | 1000 | 1000 |
| Mescaline | 1500 | 1500 |
| Metaclopramide | 1000 | 1000 |
| Methadone | 1000 | 1000 |
| Methaqualone | 1500 | 1500 |
| d,l-Methyldopa | 1000 | 1000 |

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| 1-Methyldopa | 1000 | 1000 |
| --- | --- | --- |
| Monoethylglycinexylidide (MEGX) | 1000 | 1000 |
| Morphine | 1000 | 1000 |
| Nalmefene | 20 | 20 |
| Naloxone | 500 | 500 |
| Naproxen | 1000 | 1000 |
| Nicotinic Acid | 500 | 500 |
| Nitroglycerin | 1000 | 1000 |
| Noracetylmethadol | 25 | 25 |
| 11-nor-D9-THC-9-COOH | 100 | 100 |
| Nortriptyline | 600 | 750 |
| Ofloxacin | 100 | 100 |
| Oxazepam | 300 | 300 |
| Paroxetine | 800 | 1000 |
| Phencyclidine | 1000 | 1000 |
| Phenelzine | 100 | 100 |
| 1-Phenylcyclohexylamine (PCA) | 50 | 50 |
| Phenytoin | 1000 | 1000 |
| Phthalic Acid | 1000 | 1000 |
| 1-Piperidinocyclohexane carbonitrile | 50 | 50 |
| Procainamide | 1000 | 1000 |
| Promethazine | 1000 | 1000 |
| Propoxyphene | 1000 | 1000 |
| Ranitidine | 900 | 900 |
| Sertraline | 100 | 300 |
| Scopolamine | 500 | 500 |
| Secobarbital | 1000 | 1000 |
| Thioridazine | 100 | 100 |
| Tolmetin Sodium | 2000 | 2000 |
| Tramadol | 100 | 100 |
| Trifluoperazine | 100 | 100 |
| Trimethobenzamide | 500 | 500 |
| Trimethoprim | 500 | 500 |
| Verapamil | 1000 | 1000 |
| Zidovudine (AZT) | 2000 | 2000 |
| Zolpidem | 100 | 100 |
| Diethylpropion HCl | 1000 | 1000 |
| d,l-Isoproterenol | 400 | 1000 |
| Metaproterenol | 200 | 500 |
| Methylphenidate (Ritalin®) | 1000 | 1000 |
| Phendimetrazine | 400 | 400 |
| Phenethylamine | 20 | 20 |
| Phenylephrine | 400 | 1000 |
| Propylhexedrine | 125 | 125 |
| 3-OH-Tyramine (dopamine) | 300 | 300 |

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To test for potential positive/and or negative interference from endogenous conditions the sponsor spiked each endogenous compound into a negative or positive control. The mean rate of the control with the interferent was compared to the cutoff rate. The criterion applied by the sponsor to conclude that there was or was not interference by the compound was: negative control spiked with endogenous interferents must not produce a positive result and positive control spiked with endogenous interferents must not produce a negative result.

According to these criteria, there was no change in any test samples as compared to the results of the control samples.

# At the $300\mathrm{ng / mL}$ cutoff

Negative Control $(225\mathrm{ng / mL}) = 385.5\mathrm{mA / min}$ (without interferent)

$300\mathrm{ng / mL}$ cutoff $= 420.5\mathrm{mA / min}$

| Endogenous Compound | Concentration | Mean Rate of Negative Control plus interferent |
| --- | --- | --- |
| Acetone | 1.0 g/dL | 394 mA/min |
| Ascorbic Acid | 1.5 g/dL | 389 mA/min |
| Bilirubin | 2.0 g/dL | 388 mA/min |
| Creatinine | 0.5 g/dL | 399 mA/min |
| Ethanol | 1.0 g/dL | 398 mA/min |
| Glucose | 2.0 g/dL | 399 mA/min |
| Hemoglobin | 115 mg/dL | 396 mA/min |
| Human Serum Albumin | 0.5 g/dL | 398 mA/min |
| IgG | 0.5 g/dL | 398 mA/min |
| Oxalic Acid | 0.1 g/dL | 395 mA/min |
| Riboflavin | 7.5 mg/dL | 390 mA/min |
| Sodium Chloride | 6.0 g/dL | 408 mA/min |
| Urea | 6.0 g/dL | 394 mA/min |

# At the $300\mathrm{ng / mL}$ cutoff

Positive Control $(375\mathrm{ng / mL}) = 454.6\mathrm{mA / min}$ (without interferent)

$300\mathrm{ng / mL}$ cutoff $= 420.5\mathrm{mA / min}$

| Endogenous Compound | Concentration | Mean Rate of Positive Control plus interferent |
| --- | --- | --- |
| Acetone | 1.0 g/dL | 466 mA/min |
| Ascorbic Acid | 1.5 g/dL | 465 mA/min |
| Bilirubin | 2.0 g/dL | 468 mA/min |
| Creatinine | 0.5 g/dL | 463 mA/min |
| Ethanol | 1.0 g/dL | 464 mA/min |
| Glucose | 2.0 g/dL | 467 mA/min |
| Hemoglobin | 115 mg/dL | 464 mA/min |

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| Human Serum Albumin | 0.5 g/dL | 467 mA/min |
| --- | --- | --- |
| IgG | 0.5 g/dL | 467 mA/min |
| Oxalic Acid | 0.1 g/dL | 466 mA/min |
| Riboflavin | 7.5 mg/dL | 466 mA/min |
| Sodium Chloride | 6.0 g/dL | 475 mA/min |
| Urea | 6.0 g/dL | 468 mA/min |

At the 500 ng/mL cutoff
Negative Control (375 ng/mL) = 454.6 mA/min (without interferent)
500 ng/mL cutoff = 494.3 mA/min

| Endogenous Compound | Concentration | Mean Rate of Negative Control plus interferent |
| --- | --- | --- |
| Acetone | 1.0 g/dL | 466 mA/min |
| Ascorbic Acid | 1.5 g/dL | 465 mA/min |
| Bilirubin | 2.0 g/dL | 468 mA/min |
| Creatinine | 0.5 g/dL | 463 mA/min |
| Ethanol | 1.0 g/dL | 464 mA/min |
| Glucose | 2.0 g/dL | 467 mA/min |
| Hemoglobin | 115 mg/dL | 464 mA/min |
| Human Serum Albumin | 0.5 g/dL | 467 mA/min |
| IgG | 0.5 g/dL | 467 mA/min |
| Oxalic Acid | 0.1 g/dL | 466 mA/min |
| Riboflavin | 7.5 mg/dL | 466 mA/min |
| Sodium Chloride | 6.0 g/dL | 475 mA/min |
| Urea | 6.0 g/dL | 468 mA/min |

At the 500 ng/mL cutoff
Positive Control (625 ng/mL) = 523.9 mA/min (without interferent)
500 ng/mL cutoff = 494.3 mA/min

| Endogenous Compound | Concentration | Mean Rate of Positive Control plus interferent |
| --- | --- | --- |
| Acetone | 1.0 g/dL | 529 mA/min |
| Ascorbic Acid | 1.5 g/dL | 525 mA/min |
| Bilirubin | 2.0 g/dL | 529 mA/min |
| Creatinine | 0.5 g/dL | 529 mA/min |
| Ethanol | 1.0 g/dL | 528 mA/min |
| Glucose | 2.0 g/dL | 526 mA/min |
| Hemoglobin | 115 mg/dL | 528 mA/min |
| Human Serum Albumin | 0.5 g/dL | 531 mA/min |
| IgG | 0.5 g/dL | 528 mA/min |
| Oxalic Acid | 0.1 g/dL | 526 mA/min |
| Riboflavin | 7.5 mg/dL | 526 mA/min |
| Sodium Chloride | 6.0 g/dL | 532 mA/min |
| Urea | 6.0 g/dL | 527 mA/min |

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# Testing for pH Interference at the 300 ng/mL cutoff

Negative Control (225 ng/mL) = 385.7 mA/min

300 ng/mL cutoff = 420.5 mA/min

Positive Control (375 ng/mL) = 454.6 mA/min

| pH tested | Mean Rate of the Negative Control (mA/min) | Mean Rate of the Positive Control (mA/min) |
| --- | --- | --- |
| 3.0 | 404 | 465 |
| 4.0 | 402 | 462 |
| 4.5 | 405 | 447 |
| 8.0 | 406 | 468 |
| 10.0 | 411 | 458 |
| 11.0 | 392 | 463 |

# Testing for pH Interference at the 500 ng/mL cutoff

Negative Control (375 ng/mL) = 454.6 mA/min

500 ng/mL cutoff = 494.1 mA/min

Positive Control (625 ng/mL) = 523.9 mA/min

| pH tested | Mean Rate of the Negative Control (mA/min) | Mean Rate of the Positive Control (mA/min) |
| --- | --- | --- |
| 3.0 | 465 | 531 |
| 4.0 | 462 | 531 |
| 4.5 | 447 | 547 |
| 8.0 | 468 | 526 |
| 10.0 | 458 | 529 |
| 11.0 | 463 | 521 |

There is the possibility that other substances and/or factors not listed above may interfere with the test and cause false results, e.g., technical or procedural errors.

## f. Assay cut-off:

The 500 ng/mL cutoff concentration of the assay is recommended for use by the Substance Abuse and Mental Health Services Administration (SAMHSA) for screening purposes for MDMA. The user may also choose to use a 300 ng/mL cutoff concentration, as recommended by the United Kingdom Laboratory Guidelines for Legally Defensible Workplace Drug Testing.

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Characterization of how the device performs analytically around the claimed cutoff concentration appears in the precision section, above.

2. Comparison studies:

a. Method comparison with predicate device:

Because the candidate device was compared to a reference method, GC/MS, it was not compared to a predicate device.

To evaluate the 300 ng/mL and 500 ng/mL cutoffs a total of 100 samples were evaluated by the candidate device and by GC/MS.

Sample description: for the evaluation of the 300 ng/mL cutoff, 78 unaltered clinical urine samples were evaluated. 22 additional diluted samples were also included in the study. For the evaluation of the 500 ng/mL cutoff, 77 unaltered clinical urine samples were evaluated plus an additional 23 diluted samples. The diluted samples were prepared by diluting clinical samples with high drug concentrations with drug-free urine. This was done in order to obtain samples near the cutoff concentration of the assay, because the sponsor was not able to obtain unaltered samples near the cutoff.

Sample selection: Samples were chosen such that the MDMA concentration by GC/MS would cover the assay range of the candidate device.

The study included an adequate number of samples that contained drugs near to the cutoff concentration of the assay. Approximately 10% of the study samples are evenly distributed between plus and minus 50% of the claimed cutoff concentration.

Number of study sites: one

Description of the site: Manufacturer's Research and Development facility

Operator description: Manufacturer's Research and Development staff

Number of instruments used: one

Note: due to differences in calibration curves, a semi-quantitative result in ng/mL may not always be in agreement with a qualitative result for the same sample. For example, when using the 300 ng/mL cutoff, the assay might produce a semi-quantitative result of 287 ng/mL and a qualitative result of positive.

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# Candidate Device Results vs. GC/MS Values - 300 ng/mL cutoff*

| | Positive by UK Confirmation Cutoff | Negative by UK Confirmation Cutoff |
| --- | --- | --- |
| Positive by Candidate Device | 55 | 0 |
| Negative by Candidate Device | 2 | 43 |

*Samples are classified as positive or negative based on the United Kingdom Laboratory Guidelines for Legally Defensible Workplace Drug Testing GC/MS confirmation cutoffs, which are: 200 ng/mL MDMA or 200 ng/mL MDEA or 200 ng/mL MDA.

# Candidate Device Results vs. stratified GC/MS Values - 300 ng/mL cutoff*

| Candidate Device Results | Less than half the cutoff concentration by GC/MS analysis | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration) | High Positive (greater than 50% above the cutoff concentration) |
| --- | --- | --- | --- | --- |
| Positive | 0 | 7 | 8 | 40 |
| Negative | 41 | 3 | 1 | 0 |

*Samples are classified as positive or negative determined by adding together the GC/MS concentrations for MDMA, MDA, and MDEA

% Agreement among positives is 98%

% Agreement among negatives is 86%

# Candidate Device Results vs. GC/MS Values - 500 ng/mL cutoff*

| | Positive by SAMHSA Confirmation Cutoff | Negative by SAMHSA Confirmation Cutoff |
| --- | --- | --- |
| Positive by Candidate Device | 53 | 0 |
| Negative by Candidate Device | 4 | 43 |

*Samples are classified as positive or negative based on the Proposed SAMHSA Mandatory Guidelines for Federal Workplace Drug Testing Programs confirmation cutoffs, which are: 250 ng/mL MDMA or 250 ng/mL MDEA or 250 ng/mL MDA

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Candidate Device Results vs. stratified GC/MS Values - 500 ng/mL cutoff*

| Candidate Device Results | Less than half the cutoff concentration by GC/MS analysis | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration) | High Positive (greater than 50% above the cutoff concentration) |
| --- | --- | --- | --- | --- |
| Positive | 0 | 8 | 8 | 37 |
| Negative | 43 | 1 | 3 | 0 |

*Samples are classified as positive or negative determined by adding together the GC/MS concentrations for MDMA, MDA, and MDEA

% Agreement among positives is 94%

% Agreement among negatives is 85%

b. Matrix comparison:
Not applicable.

3. Clinical studies:
a. Clinical sensitivity:
Not applicable. Clinical studies are not typically submitted for this device type.
b. Clinical specificity:
Not applicable. Clinical studies are not typically submitted for this device type.
c. Other clinical supportive data (when a and b are 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.