Immunalysis Benzodiazepines Urine Enzyme Immunoassay, Immunalysis Multi-Drug Calibrators

K151771 · Immunalysis Corporation · JXM · Jan 5, 2016 · Clinical Toxicology

Device Facts

• Record ID: K151771
• Device Name: Immunalysis Benzodiazepines Urine Enzyme Immunoassay, Immunalysis Multi-Drug Calibrators
• Applicant: Immunalysis Corporation
• Product Code: JXM · Clinical Toxicology
• Decision Date: Jan 5, 2016
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3170
• Device Class: Class 2

Intended Use

The Immunalysis Benzodiazepines Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay with a cutoff of 200ng/mL. The assay is intended for use in laboratories for the qualitative and semi-quantitative analysis of Benzodiazepines in human urine with automated clinical chemistry analyzers. This assay is calibrated against Oxazepam. This in-vitro 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 Gas Chromatography/ Mass Spectrometry (GC-MS) or permitting laboratories to establish quality control procedures. The Immunalysis Benzodiazepines Urine Enzyme Immunoassay Kit provides only a preliminary analytical test result. A more specific alternate chemical must be used in order to obtain a confirmed analytical result. GC-MS or Liquid Chromatography / Mass Spectrometry (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 Multi-Drug Calibrators are intended for in vitro diagnostic use for the calibration of assays for the analytes currently listed in the package insert: Benzoylecgonine, Morphine and Oxazepam. The calibrators are designed for prescription use with immunoassays.

Device Story

The Immunalysis Benzodiazepines Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay used in clinical laboratories to detect benzodiazepines in human urine. The device utilizes monoclonal antibodies to benzodiazepines, glucose-6-phosphate, and NAD in an antibody/substrate reagent, combined with a benzodiazepine derivative labeled with G6PDH enzyme. When benzodiazepines are present in the urine sample, they compete with the enzyme-labeled drug for antibody binding sites, resulting in a measurable change in enzyme activity. The assay is performed on automated clinical chemistry analyzers (e.g., Beckman Coulter AU 400e). Results are provided as qualitative (positive/negative relative to a 200ng/mL cutoff) or semi-quantitative. The output assists clinicians in identifying potential drug presence, which must be confirmed by GC-MS or LC/MS. The device aids in clinical decision-making regarding drug abuse testing and quality control procedures.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated over 20 days (N=80) at concentrations ±25%, ±50%, ±75%, and ±100% of the 200 ng/mL cutoff. Linearity confirmed from 100-1100 ng/mL. Method comparison with LC/MS using 86 clinical urine samples showed 98% positive agreement and 100% negative agreement. Interference testing performed for structurally related/unrelated compounds, endogenous substances, pH, and specific gravity.

Technological Characteristics

Homogeneous enzyme immunoassay; two-reagent liquid format (antibody/substrate and enzyme conjugate). Reagents contain monoclonal antibodies, G6PDH-labeled benzodiazepine derivative, G6P, NAD, Tris/HEPES buffers, and sodium azide preservative. Operates via spectrophotometric measurement at 340 nm. Designed for use on automated clinical chemistry analyzers. Storage at 2-8°C.

Indications for Use

Indicated for the qualitative and semi-quantitative analysis of benzodiazepines in human urine in laboratory settings using automated clinical chemistry analyzers. Intended for prescription use to provide preliminary analytical results requiring confirmation by GC-MS or LC/MS.

Regulatory Classification

Identification

A benzodiazepine test system is a device intended to measure any of the benzodiazepine compounds, sedative and hypnotic drugs, in blood, plasma, and urine. The benzodiazepine compounds include chlordiazepoxide, diazepam, oxazepam, chlorzepate, flurazepam, and nitrazepam. Measurements obtained by this device are used in the diagnosis and treatment of benzodiazepine use or overdose and in monitoring levels of benzodiazepines to ensure appropriate therapy.

Special Controls

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

• DRI® Benzodiazepines Assay (k930529)
• LZI Multiple Analyte Drugs of Abuse Calibrators and Controls (k051088)

Submission Summary (Full Text)

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

January 5, 2016

IMMUNALYSIS CORPORATION JOSEPH GINETE REGULATORY AFFAIRS SPECIALIST II 829 TOWNE CENTER DR POMONA CA 91767

Re: K151771

Trade/Device Name: Immunalysis Benzodiazepines Urine Enzyme Immunoassay. Immunalysis Multi-drug Calibrators Regulation Number: 21 CFR 862.3170 Regulation Name: Benzodiazepine test system Regulatory Class: II Product Code: JXM, DKB Dated: December 24, 2015 Received: December 28, 2015

Dear Mr. Ginete:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Courtney

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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## Indications for Use

510(k) Number (if known)

#### Device Name

Immunalysis Benzodiazepines Urine Enzyme Immunoassay Immunalysis Multi-Drug Calibrators

#### Indications for Use (Describe)

The Immunalysis Benzodiazepines Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay with a cutoff of 200ng/mL. The assay is intended for use in laboratories for the qualitative and semi-quantitative analysis of Benzodiazepines in human urine with automated clinical chemistry analyzers. This assay is calibrated against Oxazepam. This in-vitro 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 Gas Chromatography/ Mass Spectrometry (GC-MS) or permitting laboratories to establish quality control procedures.

The Immunalysis Benzodiazepines Urine Enzyme Immunoassay Kit provides only a preliminary analytical test result. A more specific alternate chemical must be used in order to obtain a confirmed analytical result. GC-MS or Liquid Chromatography / Mass Spectrometry (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 Multi-Drug Calibrators are intended for in vitro diagnostic use for the calibration of assays for the analytes currently listed in the package insert: Benzoylecgonine, Morphine and Oxazepam. The calibrators are designed for prescription use with immunoassays.

Type of Use (Select one or both, as applicable)

| <div> <span></span>Prescription Use (Part 21 CFR 801 Subpart D) </div> | <div> <span></span>Over-The-Counter Use (21 CFR 801 Subpart C) </div> |
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## 510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92(c).

- A. Contact Information

| 1. Manufacturer: | Immunalysis Corporation |
|-------------------------|-----------------------------------------|
| 2. Contact Name: | Joseph Ginete |
| 3. Contact Title: | Regulatory Affairs Specialist II |
| 4. Address: | 829 Towne Center Drive Pomona, CA 91767 |
| 5. Phone: | [phone] |
| 6. Fax: | [phone] |
| 7. Email: | [e-mail] |
| 8. Summary prepared on: | December 24, 2015 |

- B. Device Information
- 1. Trade Name: Immunalysis Benzodiazepine Urine Enzyme Immunoassay Immunalysis Multi-Drug Calibrators
- 2. Common Name: Immunalysis Benzodiazepine Urine Enzyme Immunoassay Immunalysis Multi-Drug Calibrators

### C. Regulatory Information

- 1. Device Classification:
- 2. Regulation Number: 21 CFR862.3170 Benzodiazepine Test System

II

21 CFR 862.3200 Clinical Toxicology Calibrator

- 3. Panel: Toxicology(91)
- JXM 4. Product Code:

DKB

- D. Legally Marketed Device to Which We are Claiming Equivalence (807.92(A)(3))

| 1. | Predicate Device: | DRI® Benzodiazepines Assay<br>LZI Multiple Analyte Drugs of Abuse Calibrators<br>and Controls |
|----|---------------------|-----------------------------------------------------------------------------------------------|
| 2. | Predicate Company: | Microgenics<br>Lin-Zhi International, Inc. |
| 3. | Predicate K Number: | K930529<br>K051088 |

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- E. Device Description
- 1. The assay consists of antibody/ substrate reagent and enzyme conjugate reagent. The antibody/ substrate reagent includes monoclonal antibodies to Benzodiazepine, glucose-6-phosphate (G6P) and nicotinamide adenine dinucleotide (NAD) in HEPES buffer with Sodium Azide as a preservative. The enzyme conjugate reagent includes Benzodiazepines derivative labeled with glucose-6-phosphate dehydrogenase (G6PDH) in HEPES buffer with Sodium Azide as a preservative.
- 2. All of the Immunalysis Multi-Drug Calibrators are liquid and ready to use. Each contains a known concentration of a specific drug analyte as a mixture.

The negative calibrator is a processed, drug-free synthetic urine matrix with sodium azide as a preservative. The Level 1, 2, 3 and 4 calibrators are prepared by spiking known concentrations of drug analyte into the negative calibrator matrix. These five calibrators (negative, Level 1, 2, 3 and 4) are sold as individual bottles. The concentration of drug analyte in the corresponding calibrators is summarized as follows:

| Table 1 Immunalysis Multi-Drug Calibrators | | | | |
|--------------------------------------------|-----------|----------|----------|-----------|
| Analyte | Level 1 | Level 2 | Level 3 | Level 4 |
| Benzoylecgonine | 150ng/mL | 300ng/mL | 500ng/mL | 1000ng/mL |
| Morphine | 100ng/mL | 300ng/mL | 500ng/mL | 1000ng/mL |
| PCP | 12.5ng/mL | 25ng/mL | 50ng/mL | 100ng/mL |
| Oxazepam | 100ng/mL | 200ng/mL | 500ng/mL | 1000ng/mL |

### F. Intended Use

- 1. Immunalysis Benzodiazepine Urine Enzyme Immunoassay
The Immunalysis Benzodiazepine Urine Enzyme Immunoassay is a homogeneous enzyme immunoassay with a cutoff of 200ng/mL. The assay is intended for use in laboratories for the qualitative and semi-quantitative analysis of Benzodiazepine in human urine with automated clinical chemistry analyzers. This assay is calibrated against Oxazepam. This in-vitro 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 Gas Chromatography/ Mass Spectrometry (GC-MS) or permitting laboratories to establish quality control procedures.

The Immunalysis Benzodiazepine 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. GC-MS or Liquid Chromatography / Mass Spectrometry (LC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to

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any drug of abuse test result, particularly when preliminary positive results are used.

- 2. Immunalysis Multi-Drug Calibrators
The Immunalysis Multi-Drug Calibrators are intended for in vitro diagnostic use for the calibration of assays for the analytes currently listed in the package insert: Benzoylecgonine, Morphine, PCP and Oxazepam. The calibrators are designed for prescription use with immunoassays.

- G. Comparison of the new device with the predicate device

| Item | Benzodiazepines Assay K930529 | Immunalysis Benzodiazepines Urine EIA |
|--------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | For the qualitative and semi-quantitative determination of the presence of Benzodiazepines in human urine at a cutoff of 200ng/mL | For the qualitative and semi-quantitative determination of the presence of Benzodiazepines in human urine at a cutoff of 200ng/mL |
| Type of Product | Analytical Reagents | Analytical Reagents |
| Measured<br>Analytes | Benzodiazepine | Benzodiazepine |
| Test Matrix | Urine | Urine |
| Cutoff Levels | 200ng/mL of Oxazepam | 200ng/mL of Oxazepam |
| Test System | Homogeneous Enzyme<br>Immunoassay | Homogeneous Enzyme Immunoassay |
| Materials | Liquid Ready-to-Use Two Reagent Assay (R1 and R2) | Antibody/Substrate Reagents and Enzyme Labeled Conjugate |
| Mass<br>Spectroscopy<br>Confirmation | Required for preliminary positive analytical results | Required for preliminary positive analytical results |
| Antibody | Sheep Polyclonal to Benzodiazepine | Monoclonal antibody to Benzodiazepines |
| Storage | 2 – 8°C until expiration date | 2 - 8°C until expiration date |

| Item | LZI Multiple Analyte K051088 | Immunalysis Multi-Drug Calibrator |
|-------------------|---------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------|
| Analyte | benzoylecgonine, d-<br>methamphetamine, methadone,<br>morphine, oxazepam,<br>secobarbital, phencyclidine,<br>propoxyphene | benzoylecgonine, morphine, PCP, oxazepam |
| Matrix | Urine | Urine |
| Calibrator Levels | 5 Levels – See Table 2 Below | 5 Levels (Negative and Level 1, 2, 3 and 4) -<br>See Device Description Table 1 |
| Storage | 2 – 8°C until expiration date | 2 - 8°C until expiration date |

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| Table 2 LZI Multiple Analyte DAU Calibrators and Controls | | | | |
|-----------------------------------------------------------|-----------|-----------|--------------|-----------|
| Multiple Analyte Calibrators | | | | |
| Analyte | Low | Cutoff | Intermediate | High |
| d-Methamphetamine | 250ng/mL | 500ng/mL | 750ng/mL | 1000ng/mL |
| Morphine | 1000ng/mL | 2000ng/mL | 4000ng/mL | 5000ng/mL |
| Phencyclidine | 12.5ng/mL | 25ng/mL | 50ng/mL | 100ng/mL |
| Benzoylecgonine | 75ng/mL | 150ng/mL | 300ng/mL | 1000ng/mL |
| Oxazepam | 100ng/mL | 200ng/mL | 500ng/mL | 1000ng/mL |
| Secobarbital | 100ng/mL | 200ng/mL | 500ng/mL | 1000ng/mL |
| Propoxyphene | 150ng/mL | 300ng/mL | 600ng/mL | 1000ng/mL |
| Methadone | 150ng/mL | 300ng/mL | 600ng/mL | 1000ng/mL |

- H. The following laboratory performance studies were performed to determine substantial equivalence of the Immunalysis Benzodiazepines Urine Enzyme Immunoassay to the predicate
- 1. Precision/ Cutoff Characterization/ Reproducibility Precision/Cutoff Characterization - Study was performed for 20 days, 2 runs per day in duplicate on drug free urine (N=80) spiked with oxazepam to concentration of ±25%, ±50%, ±75%, and ±100% of the cutoff. The spiked concentrations were confirmed by mass spectrometry (MS). The study verified that the cutoff serves as a boundary between a negative and positive interpretation of a qualitative result. The instruments used for this was Beckman Coulter AU 400e.
- a. The following is a summary table of the Qualitative Analysis for the 200ng/mL cutoff test data results.

| Table 3 - Qualitative Analysis (for 200ng/mL cutoff) | | | |
|------------------------------------------------------|-------------|---------------------|-----------------|
| Concentration (ng/mL) | % of cutoff | # of determinations | Result |
| 0 | -100% | 80 | 80 Negative |
| 50 | -75% | 80 | 80 Negative |
| 100 | -50% | 80 | 80 Negative |
| 150 | -25% | 80 | 80 Negative |
| 200 | Cutoff | 80 | 37 Neg / 43 Pos |
| 250 | +25% | 80 | 80 Positive |
| 300 | +50% | 80 | 80 Positive |
| 350 | +75% | 80 | 80 Positive |
| 400 | +100% | 80 | 80 Positive |

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| Table 4 - Semi-Quantitative Analysis (for 200ng/mL cutoff) | | | |
|------------------------------------------------------------|-------------|---------------------|-----------------|
| Concentration (ng/mL) | % of cutoff | # of determinations | Result |
| 0 | -100% | 80 | 80 Negative |
| 50 | -75% | 80 | 80 Negative |
| 100 | -50% | 80 | 80 Negative |
| 150 | -25% | 80 | 80 Negative |
| 200 | Cutoff | 80 | 34 Neg / 46 Pos |
| 250 | +25% | 80 | 80 Positive |
| 300 | +50% | 80 | 80 Positive |
| 350 | +75% | 80 | 80 Positive |
| 400 | +100% | 80 | 80 Positive |

- b. The following is a summary table of the Semi-Quantitative Analysis for the 200ng/mL cutoff test data results.
- 2. Specificity and Cross-Reactivity Various Benzodiazepines or structurally similar compounds were spiked into drug free urine at levels that will yield a result that is equivalent to the cutoff. The study verified assay performance relative to the ability of the device to exclusively determine certain drugs, in both the qualitative and semi-quantitative modes. The instrument used for this test was a Beckman Coulter AU 400e.

| ucturelly Roloted Compounds (for 200 ng/m] cutoff) - Quelitotive | |
|-----------------------------------------------------------------------------|--|
| below: | |
| a. The qualitative result summary table for the 200ng/mL cutoff is outlined | |
| | |
| | |

| Table 5 - Structurally Related Compounds (for 200 ng/mL cutoff) - Qualitative | | | |
|-------------------------------------------------------------------------------|------------------------------|--------|----------------------|
| Compound | Concentration Tested (ng/mL) | Result | Cross-Reactivity (%) |
| Oxazepam | 200 | POS | 100.0 |
| Alpha-hydroxyalprazolam | 110 | POS | 181.8 |
| Alprazolam | 120 | POS | 166.7 |
| 7-Aminoclonazepam | 100,000 | NEG | < 0.002 |
| 7-Aminoflunitrazepam | 3,500 | POS | 5.7 |
| 7-Aminonitrazepam | 35,000 | POS | 0.6 |
| Bromazepam | 750 | POS | 26.7 |
| Chlordiazepoxide | 1,600 | POS | 12.5 |
| Clorazepate | 200 | POS | 100.0 |
| Clobazam | 650 | POS | 30.8 |
| Clonazepam | 180 | POS | 111.1 |
| Demoxepam | 5,500 | POS | 3.6 |
| Desalkyflurazepam | 75 | POS | 266.7 |
| Diazepam | 100 | POS | 200.0 |
| Estazolam | 225 | POS | 88.9 |
| Flunitrazepam | 125 | POS | 160.0 |
| Flurazepam | 110 | POS | 181.8 |
| Lorazepam | 60 | POS | 333.3 |
| Lorazepam glucuronide | 180 | POS | 111.1 |
| Lormetazepam | 50 | POS | 400.0 |

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## IMMUNALYSIS

| Table 5 - Structurally Related Compounds (for 200 ng/mL cutoff) - Qualitative | | | |
|-------------------------------------------------------------------------------|------------------------------|--------|----------------------|
| Compound | Concentration Tested (ng/mL) | Result | Cross-Reactivity (%) |
| Medazepam | 500 | POS | 40.0 |
| Midazolam | 40 | POS | 500.0 |
| Nitrazepam | 700 | POS | 28.6 |
| Norchlordiazepoxide | 2,200 | POS | 9.1 |
| Nordiazepam | 180 | POS | 111.1 |
| Oxazepam glucuronide | 1,300 | POS | 15.4 |
| Prazepam | 95 | POS | 210.5 |
| Temazepam | 110 | POS | 181.8 |
| Temazepam glucuronide | 700 | POS | 28.6 |
| Triazolam | 50 | POS | 400.0 |

b. The semi-quantitative result summary table for the 200ng/mL cutoff is outlined below:

| Table 6 - Structurally Related Compounds (for 200ng/mL cutoff) - Semi-Quantitative | | | | | | |
|------------------------------------------------------------------------------------|------------------------------|--------|----------------------|--|--|--|
| Compound | Concentration Tested (ng/mL) | Result | Cross-Reactivity (%) | | | |
| Oxazepam | 200 | POS | 100.0 | | | |
| Alpha-hydroxyalprazolam | 110 | POS | 181.8 | | | |
| Alprazolam | 120 | POS | 166.7 | | | |
| 7-Aminoclonazepam | 100,000 | NEG | < 0.002 | | | |
| 7-Aminoflunitrazepam | 3,500 | POS | 5.7 | | | |
| 7-Aminonitrazepam | 35,000 | POS | 0.6 | | | |
| Bromazepam | 750 | POS | 26.7 | | | |
| Chlordiazepoxide | 1,600 | POS | 12.5 | | | |
| Clorazepate | 200 | POS | 100.0 | | | |
| Clobazam | 650 | POS | 30.8 | | | |
| Clonazepam | 180 | POS | 11.1 | | | |
| Demoxepam | 5,500 | POS | 3.6 | | | |
| Desalkyflurazepam | 75 | POS | 266.7 | | | |
| Diazepam | 100 | POS | 200.0 | | | |
| Estazolam | 225 | POS | 88.9 | | | |
| Flunitrazepam | 125 | POS | 160.0 | | | |
| Flurazepam | 110 | POS | 181.8 | | | |
| Lorazepam | 60 | POS | 333.3 | | | |
| Lorazepam glucuronide | 180 | POS | 111.1 | | | |
| Lormetazepam | રે0 | POS | 400.0 | | | |
| Medazepam | 500 | POS | 40.0 | | | |
| Midazolam | 40 | POS | 500.0 | | | |
| Nitrazepam | 700 | POS | 28.6 | | | |
| Norchlordiazepoxide | 2,200 | POS | 9.1 | | | |
| Nordiazepam | 180 | POS | 11.1 | | | |
| Oxazepam glucuronide | 1,300 | POS | 15:4 | | | |
| Prazepam | તેર | POS | 210.5 | | | |
| Temazepam | 110 | POS | 181.8 | | | |
| Temazepam glucuronide | 700 | POS | 28.6 | | | |
| Triazolam | ર૦ | POS | 400.0 | | | |

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## IMMUNALYSIS

- 3. Interference Structurally unrelated compounds were evaluated in qualitative and semi-quantitative modes by spiking the potential interferent into drug free urine containing oxazepam at ±25% of the cutoff. All potential interferents analyzed verified that assay performance is unaffected by externally ingested compounds. The instrument used for this test was a Beckman Coulter AU 400e.
- a. The following is a table of the structurally non-similar compounds for the 200ng/mL cutoff:

| Table 7 - Structurally Unrelated Compounds (for 200ng/mL cutoff) | | | | | |
|------------------------------------------------------------------|-------------------|------------------------|-------------------|------------------------|-------------------|
| | Concentration | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| Compound | Tested<br>(ng/mL) | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| 4-Bromo-<br>2,5,Dimethoxyphenethylamine | 100,000 | Negative | Negative | Positive | Positive |
| 6-Acetylcodeine | 100,000 | Negative | Negative | Positive | Positive |
| 6-Acetylmorphine | 100,000 | Negative | Negative | Positive | Positive |
| Acetaminophen | 500,000 | Negative | Negative | Positive | Positive |
| Acetylsalicylic Acid | 500,000 | Negative | Negative | Positive | Positive |
| Amitriptyline | 100,000 | Negative | Negative | Positive | Positive |
| Amobarbital | 100,000 | Negative | Negative | Positive | Positive |
| S-(+) Amphetamine | 100,000 | Negative | Negative | Positive | Positive |
| Benzoylecgonine | 500,000 | Negative | Negative | Positive | Positive |
| Benzylpiperazine | 100,000 | Negative | Negative | Positive | Positive |
| Buprenorphine | 100,000 | Negative | Negative | Positive | Positive |
| Bupropion | 100,000 | Negative | Negative | Positive | Positive |
| Butabarbital | 100,000 | Negative | Negative | Positive | Positive |
| Caffeine | 500,000 | Negative | Negative | Positive | Positive |
| Carbamazepine | 100,000 | Negative | Negative | Positive | Positive |
| Chlorpromazine | 100,000 | Negative | Negative | Positive | Positive |
| cis-Tramadol | 100,000 | Negative | Negative | Positive | Positive |
| Clomipramine | 100,000 | Negative | Negative | Positive | Positive |
| Cannabidiol | 100,000 | Negative | Negative | Positive | Positive |
| Cannabinol | 100,000 | Negative | Negative | Positive | Positive |
| Carisoprodol | 100,000 | Negative | Negative | Positive | Positive |
| Cocaine | 100,000 | Negative | Negative | Positive | Positive |
| Codeine | 100,000 | Negative | Negative | Positive | Positive |
| Cotinine | 100,000 | Negative | Negative | Positive | Positive |
| Cyclobenzaprine | 100,000 | Negative | Negative | Positive | Positive |
| Delta-9-THC | 100,000 | Negative | Negative | Positive | Positive |
| Desipramıne | 100,000 | Negative | Negative | Positive | Positive |
| N-desmethyltapentadol | 100,000 | Negative | Negative | Positive | Positive |
| Dextromethorphan | 100,000 | Negative | Negative | Positive | Positive |
| Dihydrocodeine | 100,000 | Negative | Negative | Positive | Positive |
| Diphenhydramine | 500,000 | Negative | Negative | Positive | Positive |
| Doxepin | 100,000 | Negative | Negative | Positive | Positive |
| Ecgonine | 100,000 | Negative | Negative | Positive | Positive |
| Ecgonine methyl ester | 100,000 | Negative | Negative | Positive | Positive |
| EDDP | 100,000 | Negative | Negative | Positive | Positive |
| | Concentration | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| Compound | Tested<br>(ng/mL) | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| 1R,2S(-)-Ephedrine | 100,000 | Negative | Negative | Positive | Positive |
| 1S,2R(+)-Ephedrine | 100,000 | Negative | Negative | Positive | Positive |
| Ethyl β-D-glucuronide | 100,000 | Negative | Negative | Positive | Positive |
| Ethylmorphine | 100,000 | Negative | Negative | Positive | Positive |
| Fenfluramine | 100,000 | Negative | Negative | Positive | Positive |
| Fentanyl | 100,000 | Negative | Negative | Positive | Positive |
| Fluoxetine | 100,000 | Negative | Negative | Positive | Positive |
| Heroin | 100,000 | Negative | Negative | Positive | Positive |
| Hexobarbital | 100,000 | Negative | Negative | Positive | Positive |
| Hydrocodone | 100,000 | Negative | Negative | Positive | Positive |
| Hydromorphone | 100,000 | Negative | Negative | Positive | Positive |
| 11-hydroxy-delta-9-THC | 100,000 | Negative | Negative | Positive | Positive |
| Ibuprofen | 100,000 | Negative | Negative | Positive | Positive |
| Imipramine | 100,000 | Negative | Negative | Positive | Positive |
| Ketamine | 100,000 | Negative | Negative | Positive | Positive |
| Lamotrigine | 100,000 | Negative | Negative | Positive | Positive |
| Levorphanol Tartrate | 100,000 | Negative | Negative | Positive | Positive |
| Lidocaine | 100,000 | Negative | Negative | Positive | Positive |
| LSD | 100,000 | Negative | Negative | Positive | Positive |
| Maprotiline | 100,000 | Negative | Negative | Positive | Positive |
| (+)-MDA | 100,000 | Negative | Negative | Positive | Positive |
| MDEA | 100,000 | Negative | Negative | Positive | Positive |
| MDMA | 100,000 | Negative | Negative | Positive | Positive |
| Meperidine | 100,000 | Negative | Negative | Positive | Positive |
| Meprobamate | 100,000 | Negative | Negative | Positive | Positive |
| Methadone | 500,000 | Negative | Negative | Positive | Positive |
| S(+)-Methamphetamine | 100,000 | Negative | Negative | Positive | Positive |
| Methaquolone | 100,000 | Negative | Negative | Positive | Positive |
| Methylphenidate | 100,000 | Negative | Negative | Positive | Positive |
| Morphine | 100,000 | Negative | Negative | Positive | Positive |
| Morphine-3 -glucuronide | 100,000 | Negative | Negative | Positive | Positive |
| Morphine-6 -glucuronide | 100,000 | Negative | Negative | Positive | Positive |
| Nalorphine | 100,000 | Negative | Negative | Positive | Positive |
| Naloxone | 100,000 | Negative | Negative | Positive | Positive |
| Naltrexone | 100,000 | Negative | Negative | Positive | Positive |
| Norbuprenorphine | 100,000 | Negative | Negative | Positive | Positive |
| Norcodeine | 100,000 | Negative | Negative | Positive | Positive |
| Normorphine | 100,000 | Negative | Negative | Positive | Positive |
| Norpropoxyphene | 100,000 | Negative | Negative | Positive | Positive |
| Norpseudoephedrine | 100,000 | Negative | Negative | Positive | Positive |
| Nortriptyline | 100,000 | Negative | Negative | Positive | Positive |
| Oxycodone | 100,000 | Negative | Negative | Positive | Positive |
| Oxymorphone | 100,000 | Negative | Negative | Positive | Positive |
| Table 7 - Structurally Unrelated Compounds (for 200ng/mL cutoff) | | | | | |
| | Concentration | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| Compound | Tested<br>(ng/mL) | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| PCP | 100,000 | Negative | Negative | Positive | Positive |
| Pentazocine | 100,000 | Negative | Negative | Positive | Positive |
| Pentobarbital | 100,000 | Negative | Negative | Positive | Positive |
| Phentermine | 100,000 | Negative | Negative | Positive | Positive |
| Phenobarbital | 100,000 | Negative | Negative | Positive | Positive |
| Phenylephedrine | 100,000 | Negative | Negative | Positive | Positive |
| Phenylpropanolamine | 100,000 | Negative | Negative | Positive | Positive |
| Phenytoin | 100,000 | Negative | Negative | Positive | Positive |
| PMA | 100,000 | Negative | Negative | Positive | Positive |
| Propoxyphene | 100,000 | Negative | Negative | Positive | Positive |
| Propranolol | 100,000 | Negative | Negative | Positive | Positive |
| Protriptyline | 100,000 | Negative | Negative | Positive | Positive |
| R,R(-)-Pseudoephedrine | 100,000 | Negative | Negative | Positive | Positive |
| S,S(+)-Pseudoephedrine | 100,000 | Negative | Negative | Positive | Positive |
| Ranitidine | 100,000 | Negative | Negative | Positive | Positive |
| Ritalinic Acid | 100,000 | Negative | Negative | Positive | Positive |
| Salicylic Acid | 100,000 | Negative | Negative | Positive | Positive |
| Secobarbial | 100,000 | Negative | Negative | Positive | Positive |
| Sertraline | 100,000 | Negative | Negative | Positive | Positive |
| Sufentanil Citrate | 100,000 | Negative | Negative | Positive | Positive |
| 11-nor-9 carboxy THC | 100,000 | Negative | Negative | Positive | Positive |
| Theophylline | 100,000 | Negative | Negative | Positive | Positive |
| Thioridazine | 100,000 | Negative | Negative | Positive | Positive |
| Trifluoromethylphenyl-piperazine | 100,000 | Negative | Negative | Positive | Positive |
| Trimipramine | 100,000 | Negative | Negative | Positive | Positive |
| Trazodone | 100,000 | Negative | Negative | Positive | Positive |
| Venlafaxine | 100,000 | Negative | Negative | Positive | Positive |
| Zolpidem Tartrate | 100,000 | Negative | Negative | Positive | Positive |

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b. Endogenous compounds interference was evaluated in qualitative and semi-quantitative modes by spiking the potential interferent into drug free urine containing oxazepam at ± 25% of the cutoff. All potential interferents analyzed verified that assay performance is unaffected by internally existing physiological conditions. The instrument used for this test was a Beckman Coulter AU 400e. The following is a summary table of the endogenous compounds results for the 200ng/mL cutoff:

| Table 8 - Endogenous Compounds (for 200ng/mL cutoff) | | | | | |
|------------------------------------------------------|------------------------------------|------------------------|-------------------|------------------------|-------------------|
| Compound | Concentration<br>Tested<br>(ng/mL) | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| | | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| Acetone | 1.0 g/dL | Negative | Negative | Positive | Positive |
| Ascorbic Acid | 1.5 g/dL | Negative | Negative | Positive | Positive |
| Bilirubin | 0.002 g/dL | Negative | Negative | Positive | Positive |

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| Table 8 - Endogenous Compounds (for 200ng/mL cutoff) | | | | | |
|------------------------------------------------------|-------------------|------------------------|-------------------|------------------------|-------------------|
| | Concentration | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| Compound | Tested<br>(ng/mL) | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| Creatinine | 0.5 g/dL | Negative | Negative | Positive | Positive |
| Ethanol | 1.0 g/dL | Negative | Negative | Positive | Positive |
| Galactose | 0.01 g/dL | Negative | Negative | Positive | Positive |
| y-Globulin | 0.5 g/dL | Negative | Negative | Positive | Positive |
| Glucose | 2.0 g/dL | Negative | Negative | Positive | Positive |
| Hemoglobin | 0.115 g/dL | Negative | Negative | Positive | Positive |
| Human Serum Albumin | 0.5 g/dL | Negative | Negative | Positive | Positive |
| Oxalic Acid | 0.1 g/dL | Negative | Negative | Positive | Positive |
| Riboflavin | 0.0075 g/dL | Negative | Negative | Positive | Positive |
| Sodium Azide | 1% w/v | Negative | Negative | Positive | Positive |
| Sodium Chloride | 6.0 g/dL | Negative | Negative | Positive | Positive |
| Sodium Fluoride | 1% w/v | Negative | Negative | Positive | Positive |
| Urea | 6.0 g/dL | Negative | Negative | Positive | Positive |

- c. Boric Acid interference was also evaluated at a concentration of 1% w/v, at ±25% and ±50%of the cutoff, in both the qualitative and semiquantitative modes. The following is a summary table of the test results at ±25% of the assay cutoff of 200ng/mL:

| Table 9 - Boric Acid (for 200ng/mL cutoff) | | | | | |
|--------------------------------------------|--------------------------|------------------------|-------------------|------------------------|-------------------|
| Compound | Concentration<br>(ng/mL) | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| | Tested | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| Boric Acid | 1% w/v | Negative | Negative | Negative | Negative |

d. The following is a summary table of the test results at ±50% of the assay cutoff of 200ng/mL:

| Compound | Concentration Tested (ng/mL) | -50% Cutoff (100ng/mL) | | +50% Cutoff (300ng/mL) | |
|------------|------------------------------|------------------------|----------------------------|------------------------|----------------------------|
| Boric Acid | 1% w/v | Qualitative Negative | Semi-Quantitative Negative | Qualitative Negative | Semi-Quantitative Negative |

e. Boric Acid at a concentration of 1% w/v was found to cause false negative results at ±25% and ±50% ng/mL of the cutoff in both the qualitative and semi-quantitative modes. The following statement is provided in 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.

- f. To evaluate potential interference from th 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 oxazepam at ±25% of the 200ng/mL cutoff. No positive or negative interference was observed at urine pH values ranging from 3.0 to 11.0 for

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| Table 11 - Effect of pH (for 200ng/mL cutoff) | | | | | |
|-----------------------------------------------|-------|------------------------|-------------------------------|------------------------|-------------------------------|
| Test Parameter | Value | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| | | | Qualitative Semi-Quantitative | | Qualitative Semi-Quantitative |
| pH | 3.0 | NEG | NEG | POS | POS |
| pH | 4.0 | NEG | NEG | POS | POS |
| pH | 5.0 | NEG | NEG | POS | POS |
| pH | 6.0 | NEG | NEG | POS | POS |
| pH | 7.0 | NEG | NEG | POS | POS |
| pH | 8.0 | NEG | NEG | POS | POS |
| pH | 9.0 | NEG | NEG | POS | POS |
| pH | 10.0 | NEG | NEG | POS | POS |
| pH | 11.0 | NEG | NEG | POS | POS |

each test mode. The following is a summary table of the effect of pH results for the 200ng/mL cutoff:

g. To evaluate potential interference from the specific gravity of urine, device performance in the qualitative and semi-quantitative modes was tested using a range of physiologically relevant 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 oxazepam at ±25% of the 200ng/mL cutoff. No positive or negative interference was observed at urine specific gravity values ranging from 1.000 to 1.030 for each test mode. The following is a summary table of the effect of specific gravity results for the 200ng/mL cutoff:

| Table 12 - Effect of Specific Gravity (for 200ng/mL cutoff) | | | | | |
|-------------------------------------------------------------|-------|------------------------|-------------------|------------------------|-------------------|
| Test Parameter | Value | -25% Cutoff (150ng/mL) | | +25% Cutoff (250ng/mL) | |
| | | Qualitative | Semi-Quantitative | Qualitative | Semi-Quantitative |
| Specific Gravity | 1.000 | NEG | NEG | POS | POS |
| Specific Gravity | 1.002 | NEG | NEG | POS | POS |
| Specific Gravity | 1.005 | NEG | NEG | POS | POS |
| Specific Gravity | 1.010 | NEG | NEG | POS | POS |
| Specific Gravity | 1.015 | NEG | NEG | POS | POS |
| Specific Gravity | 1.020 | NEG | NEG | POS | POS |
| Specific Gravity | 1.025 | NEG | NEG | POS | POS |
| Specific Gravity | 1.030 | NEG | NEG | POS | POS |

- 4. Linearity/ Recovery A linearity study in the semi-quantitative mode was conducted by spiking a drug free urine pool with a high concentration of oxazepam as a high value specimen. Additional pools were made by serially diluting the high value specimen with drug free urine to achieve concentrations ranging from 100ng/mL to 1100ng/mL. Each pool was tested in triplicate to calculate the mean concentration values that were used to calculate drug recovery. The instrument used for this test was a Beckman Coulter AU 400e.

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| Table 13 - Linearity/ Recovery | | |
|--------------------------------|----------------------------|--------------|
| Expected Concentration (ng/mL) | Mean Concentration (ng/mL) | Recovery (%) |
| 100 | 94.2 | 94.2 |
| 200 | 213.7 | 106.8 |
| 300 | 313.9 | 104.6 |
| 400 | 389.7 | 97.4 |
| 500 | 511.6 | 102.3 |
| 600 | 637.2 | 106.2 |
| 700 | 693.2 | 99.0 |
| 800 | 820.7 | 102.6 |
| 900 | 907.6 | 100.8 |
| 1000 | 1025.3 | 102.5 |
| 1100 | 1061.1 | 96.5 |

a. The following is a summary table of the linearity/recovery:

- 5. Method Comparison Eighty unaltered, anonymous and discarded clinical urine samples obtained from clinical testing laboratories were analyzed for benzidiazepines with the candidates on a Beckman Coulter AU 400e clinical chemistry analyzer and with LC.MS. Results were obtained in both qualitative and semi-quantitative modes.
- a. The following is a comparison table of qualitative assay performance for the 200ng/mL cutoff:

# Table 14 - Method Comparison (for 200ng/mL cutoff) - Qualitative

| | LC/MS Confirmation | | |
|----------------|--------------------|-----|----|
| | (+) | (-) | |
| Test<br>Device | (+) | 42 | 0 |
| | (-) | 1 | 43 |

- b. The following is a summary table of qualitative assay performance for the 200ng/mL cutoff:

| Table 15 - Assay Performance verified by LC/MS – 200ng/mL Cutoff | | | | | |
|------------------------------------------------------------------|-------------------------------|-----------------|-----------------|-------------|---------------|
| Type | Benzodiazepines Concentration | | | | Agreement (%) |
| | < 100ng/mL | 100 ~ 199 ng/mL | 200 ~ 300 ng/mL | > 300 ng/mL | |
| Qualitative/<br>Positive | 0 | 0 | 4 | 38 | 100 |
| Qualitative/<br>Negative | 36 | 7 | 1 | 0 | 98 |

c. The following is a comparison table of semi-quantitative assay performance for the 200ng/mL cutoff:

### Table 16 - Method Comparison (for 200ng/mL cutoff) - Semi-Quantitative LC/MS Confirmation

| LC/MS Confirmation | | | |
|--------------------|-----|-----|---|
| | (+) | (-) | |
| Test<br>Device | (+) | 42 | 0 |
| (-) | 1 | 43 | |

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| Type | Benzodiazepines Concentration | | | Agreement (%) | |
|------------------------------|-------------------------------|-----------------|-----------------|---------------|-----|
| | < 100ng/mL | 100 ~ 199 ng/mL | 200 ~ 300 ng/mL | > 300 ng/mL | |
| Semi-Quantitative/ Positive | 0 | 0 | 4 | 38 | 100 |
| Semi-Quantitative / Negative | 36 | 7 | 1 | 0 | 98 |

- d. The following is a summary table of semi-quantitative assay performance for the 200ng/mI_cutoff.
## 6. Immunalysis Multi-Drug Calibrators Analytical Performance

- a. Traceability all components of the calibrators have been traced to a commercially available oxazepam solution.
- b.Closed Vial Stability (Accelerated) A closed vial stability study was performed at 25℃ to establish the initial vial expiration dating. All calibrator levels (1, 2, 3, and 4) were within specifications for Day 0, 8, 16, 24, 32, and 40. This accelerated stability study was performed to establish initial expiration dating. The stability study supported an initial expiration date of 12 months after testing on LC/MS. Real time stability studies are ongoing.
- c. Open Vial Stability An open vial stability study was performed at 5°C to establish the initial open vial expiration dating on LC/MS. All calibrator levels (1, 2, 3, and 4) were within specifications for Day 0, 19, 26, 33, 41, and 60. This stability study supported an initial open vial expiration date of 60 days.
- d. Value Assignment Calibrators are manufactured and are tested by mass spectrometry. The negative calibrator is a processed, drug free urine matrix. The standard is compared to a reference negative standard to ensure that it is free of analyte. The non-zero calibrators are prepared by spiking a known concentration of oxazepam in the negative calibrator matrix. If any of the analytes are not of the acceptable range, then the calibrator is adjusted and re-tested. Values are assigned to the calibrators once the mass spectrometry results are within the acceptable ranges.
- I. Proposed Labeling

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

- J. Conclusion
The information provided in this pre-market notification demonstrates that the Immunalysis Benzodiazepines Urine Enzyme Immunoassay is substantially equivalent to the legally marketed predicate device for its general intended use.