(241 days)
The CEDIA™ Benzodiazepine Assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi-quantitative determination of benzodiazepines in human urine at a cutoff concentration of 200 ng/mL.
The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The assay provides only a preliminary analytical test result. A more specific alternative chemical must be used to obtain a confirmed analytical result. Gas chromatography / Mass spectrometry (GC/MS) or Liquid chromatography/ tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.
Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
CEDIA™ technology uses recombinant DNA technology to produce a unique homogeneous enzyme immunoassay system. The assay is based on the bacterial enzyme ß-galactosidase, which has been genetically engineered into two inactive fragments, Enzyme acceptor (EA) and Enzyme Donor (ED). These fragments spontaneously re-associate to form fully active enzyme that, in the assay format, cleaves a substrate. This generates a color change that can be measured spectrophotometrically.
The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include sheep polyclonal anti-benzodiazepine antibody, recombinant microbial "enzyme donor'' - benzodiazepine conjugate, "enzyme acceptor", chlorophenol red ß-Dgalactopyranoside, stabilizers and preservatives. Add ß-glucuronidase enzyme to the reconstituted EA solution before using the assay. All specimens must be tested with ßglucuronidase enzyme. This enzyme will hydrolyze the glucuronidated metabolites of benzodiazepines in the samples, thereby enabling the detection of benzodiazepine glucuronides.
The provided text describes the analytical performance of the CEDIA™ Benzodiazepine Assay, an in vitro diagnostic device, and does not contain information related to an AI/ML-based device. Therefore, a direct response to the request for acceptance criteria and study details for an AI/ML device is not possible based on the provided text.
However, I can extract and present the acceptance criteria and the reported device performance for this in vitro diagnostic device based on the provided document.
Acceptance Criteria and Study for CEDIA™ Benzodiazepine Assay (In Vitro Diagnostic Device)
The CEDIA™ Benzodiazepine Assay is an in vitro diagnostic device for the qualitative and/or semi-quantitative determination of benzodiazepines in human urine. The document details its analytical performance studies to establish substantial equivalence to a predicate device, not an AI/ML-based system.
Here's a summary of the acceptance criteria (implied by the reported precision and accuracy goals) and the reported device performance from the provided document:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined acceptance criteria in terms of numerical thresholds for precision, accuracy, or other analytical performance metrics. Instead, it presents results intended to demonstrate that the device performs adequately and is substantially equivalent to a predicate device. For the purpose of this response, I will interpret the reported performance as meeting the implicit acceptance criteria for an in vitro diagnostic assay of this type.
Cut-off Concentration: 200 ng/mL
| Performance Metric | Implied Acceptance Criteria (Based on expected IVD performance) | Reported Device Performance |
|---|---|---|
| Precision | Qualitative Mode: Samples significantly below cutoff should be consistently negative. Samples significantly above cutoff should be consistently positive. Samples around cutoff show expected distribution. | Spiked Concentration 150 ng/mL (-25% of Cutoff): 80/80 Negative Spiked Concentration 200 ng/mL (Cutoff): 6/74 Negative/Positive (i.e., 6 negative, 74 positive) Spiked Concentration 250 ng/mL (+25% of Cutoff): 0/80 Negative/Positive (i.e., 0 negative, 80 positive) |
| Semi-Quantitative Mode: Similar to qualitative, with additional expectation of quantitative consistency. | Spiked Concentration 150 ng/mL (-25% of Cutoff): 79/1 Negative/Positive (i.e., 79 negative, 1 positive) Spiked Concentration 200 ng/mL (Cutoff): 1/79 Negative/Positive (i.e., 1 negative, 79 positive) Spiked Concentration 250 ng/mL (+25% of Cutoff): 0/80 Negative/Positive (i.e., 0 negative, 80 positive) | |
| Spike Recovery (Qualitative) | Samples spiked below cutoff should be negative; samples spiked above cutoff should be positive. | 150 ng/mL (below C/O): All 20 replicates Negative 250 ng/mL (above C/O): All 20 replicates Positive |
| Analytical Recovery and Linearity | Average recovery should be close to 100% within the assay range (e.g., 90-110%). | Range of Recovery: 95.2 – 107.8% (for expected concentrations from 100 ng/mL to 800 ng/mL) |
| Method Comparison and Accuracy (vs. LC-MS/MS) | High concordance (agreement) with the reference method (LC-MS/MS), particularly for samples clearly positive or negative. | Overall Concordance: 97% in Qualitative mode (for 200 ng/mL cutoff)96% in Semi-Quantitative mode (for 200 ng/mL cutoff)Qualitative Mode:Agreement among Positives: 100% (68/68)Agreement among Negatives: 93% (56/60)Semi-Quantitative Mode:Agreement among Positives: 99% (67/68)Agreement among Negatives: 93% (56/60) |
| Specificity (Cross-reactivity) | Minimal to no interference from structurally unrelated compounds. Relevant benzodiazepines and metabolites should show expected reactivity. | Structurally unrelated compounds (e.g., Acetaminophen, Amphetamine, Caffeine, Codeine etc. at high concentrations up to 100,000 ng/mL) showed no significant cross-reactivity, maintaining expected results for spiked low/high controls. Data provided for 30+ benzodiazepines and metabolites, showing varying cross-reactivity percentages. |
| Interference (pH, Endogenous/Exogenous Substances) | No interference from common physiological substances or varying pH levels. | Various compounds (e.g., Acetone, Ascorbic Acid, Glucose, Hemoglobin, Human Serum Albumin, Urea) and pH levels (3-11) showed no interference, maintaining expected results for spiked low/high controls. |
| Specific Gravity Interference | No interference from varying specific gravity of urine samples. | Urine samples with specific gravity from 1.002 to 1.029 showed no interference, maintaining expected results for spiked low/high controls. |
2. Sample sizes used for the test set and the data provenance
-
Precision Study:
- Samples were tested in replicates of 2, twice per day for 20 days.
- Total n=80 for each spiked concentration level (0 to 400 ng/mL).
- Data Provenance: Not explicitly stated, but the study was performed at the manufacturer's site. "Drug free urine" was used for spiking. It is implied to be laboratory-controlled samples, not patient data from a specific country or retrospective/prospective collection.
-
Spike Recovery Study:
- 20 replicates for each concentration (150 ng/mL and 250 ng/mL).
- Data Provenance: "Drug free urine" was used for spiking. Implied laboratory-controlled samples.
-
Method Comparison and Accuracy Study:
- Sample Size: One hundred and twenty-eight (128) samples.
- Data Provenance: Not explicitly stated. The samples were "treated with ß-glucuronidase reagent prior to analysis". This suggests potentially patient-derived urine samples, but their origin (country, retrospective/prospective collection) is not mentioned.
-
Specificity (Cross-reactivity) & Interference Studies:
- Samples made by adding known amounts of compounds to "drug-free negative urine" or "Oxazepam spiked" urine.
- Data Provenance: Laboratory-controlled samples.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
Not applicable for this type of in vitro diagnostic device study. The "ground truth" (or reference method) for the method comparison study was established by Liquid Chromatography/tandem mass spectrometry (LC-MS/MS), which is a highly accurate chemical analytical method. It does not involve human expert interpretation in the way AI/ML systems for medical imaging, for example, would.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
Not applicable. Ground truth was established by LC-MS/MS, an objective chemical analytical method, not by human expert consensus requiring adjudication.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
Not applicable. This is an in vitro diagnostic assay, not an AI/ML-based device that would assist human readers/operators in interpretation. Its performance is measured directly against a reference analytical method.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, in the context of an IVD, the CEDIA™ Benzodiazepine Assay's performance is inherently standalone in the sense that the assay's output (positive/negative/concentration) is directly measured and compared against the LC-MS/MS reference method. There is no human interpretative step within the 'device' itself, nor is its primary purpose to augment human interpretation.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The ground truth for the method comparison study was Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) results, which is a gold standard chemical analytical method for drug concentration determination.
8. The sample size for the training set
Not applicable. This document describes an in vitro diagnostic assay, which is a chemical and biological reagent-based system, not an AI/ML algorithm that requires a "training set" in the computational sense. Its formulation and calibration are based on chemical principles and standard laboratory practices.
9. How the ground truth for the training set was established
Not applicable, as there is no "training set" for this type of device. The assay is developed and optimized as a chemical system.
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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
December 9, 2019
Microgenics Corporation Minoti Patel Director, Regulatory Affairs 45600 Kato Road Fremont, CA 94538
Re: K190968
Trade/Device Name: CEDIA™ Benzodiazepine Assay Regulation Number: 21 CFR 862.3170 Regulation Name: Benzodiazepine Test System Regulatory Class: Class II Product Code: JXM Dated: October 21, 2019 Received: October 22, 2019
Dear Minoti Patel:
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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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
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requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Marianela Perez-Torres, Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known)
Device Name CEDIATM Benzodiazepine Assay
Indications for Use (Describe)
The CEDIA™ Benzodiazepine Assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semiquantitative determination of benzodiazepines in human urine at a cutoff concentration of 200 ng/mL.
The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The assay provides only a preliminary analytical test result. A more specific alternative chemical must be used to obtain a confirmed analytical result. Gas chromatography / Mass spectrometry (GC/MS) or Liquid chromatography/ tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.
Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
Type of Use (Select one or both, as applicable)
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) | ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
|---|---|
| ---------------------------------------------------------------------------------------------------------- | --------------------------------------------------------------------------------------------------------- |
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1. 510(k) Summary
K190968
This 510(k) Summary of Safety and Effectiveness is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.
A. Device Information
| Category | Comments |
|---|---|
| Sponsor: | Microgenics CorporationThermo Fisher Scientific46500 Kato RoadFremont, CA 94538Phone: 510-979-5000FAX: 510-979-5002 |
| Correspondent ContactInformation: | Minoti PatelDirector, Regulatory AffairsEmail: Minoti.patel@thermofisher.comPhone: 510-979-5000FAX: 510-979-5002 |
| Device Common Name: | Benzodiazepine Enzyme Immunoassay |
| Trade or Proprietary Name | CEDIATM Benzodiazepine Assay |
| Candidate Device ProductCode, Classification,Classification Name &Panel | JXM, Class II, 21 CFR 862. 3170 – BenzodiazepineTest System, 91 – Toxicology |
Predicate Device Information:
| Predicate Device: | CEDIA™ DAU Benzodiazepine Assay |
|---|---|
| Predicate Device Manufacturer: | Microgenics Corp. |
| Predicate Device Premarket Notification #: | K962734 |
B. Date Summary Prepared December 09, 2019
C. Description of Device
CEDIA™ technology uses recombinant DNA technology to produce a unique homogeneous enzyme immunoassay system. The assay is based on the bacterial enzyme ß-galactosidase, which has been genetically engineered into two inactive fragments, Enzyme acceptor (EA) and Enzyme Donor (ED). These fragments spontaneously re-associate to form fully active enzyme that, in the assay format, cleaves a substrate. This generates a color change that can be measured spectrophotometrically.
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The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include sheep polyclonal anti-benzodiazepine antibody, recombinant microbial "enzyme donor'' - benzodiazepine conjugate, "enzyme acceptor", chlorophenol red ß-Dgalactopyranoside, stabilizers and preservatives. Add ß-glucuronidase enzyme to the reconstituted EA solution before using the assay. All specimens must be tested with ßglucuronidase enzyme. This enzyme will hydrolyze the glucuronidated metabolites of benzodiazepines in the samples, thereby enabling the detection of benzodiazepine glucuronides.
D. Intended Use
The CEDIA™ Benzodiazepine Assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi-quantitative determination of benzodiazepines in human urine at a cutoff concentration of 200 ng/mL.
The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.
The 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) or Liquid chromatography/tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.
Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.
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E. Comparison to Predicate Device
| Characteristic | Candidate Device:CEDIATM Benzodiazepine Assay | Predicate Device:CEDIATM DAU Benzodiazepine(K962734) |
|---|---|---|
| Intended Use | The CEDIATM BenzodiazepineAssay is a homogeneous enzymeimmunoassay intended for thequalitative and/or semi-quantitative determination ofbenzodiazepines in human urineat a cutoff concentration of 200ng/mL.The semi-quantitative mode is forthe purpose of enablinglaboratories to determine anappropriate dilution of thespecimen for confirmation by aconfirmatory method such asLiquid Chromatography/tandemmass spectrometry (LC-MS/MS)or permitting laboratories toestablish quality controlprocedures.The assay provides only apreliminary analytical testresult. A more specificalternative chemical methodmust be used to obtain aconfirmed analytical result. Gaschromatography / Massspectrometry (GC/MS) or Liquidchromatography/tandem massspectrometry (LC-MS/MS) is thepreferred confirmatory method. | Same |
| Operating Principle(Technology) | CEDIATM | Same |
| Characteristic | Candidate Device:CEDIA™ Benzodiazepine Assay | Predicate Device:CEDIA™ DAU Benzodiazepine(K962734) |
| Measured Analyte | Benzodiazepines and theirmetabolites | Same |
| Test Matrix | Urine | Same |
| Cutoff Levels | HS 200 ng/mL | Same |
| Methodology | Homogeneous EnzymeImmunoassay | Same |
| Reagents Form | EA and ED: Lyophilized(Reconstitution Required)EARB and EDRB liquid ready-to-use. | Same |
| Antibody | Sheep polyclonal antibodies | Same |
| Storage | 2–8 °C until expiration date | Same |
| Principal Operator | Trained professionals | Same |
| Calibrator Analyte | Oxazepam | Nitrazepam |
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F. Test Principle
The CEDIA™ Benzodiazepine Assay uses recombinant DNA technology (US Patent No. 4708929) to produce a unique homogeneous enzyme immunoassay system. This assay is based on the bacterial enzyme ß-galactosidase, which has been genetically engineered into two inactive fragments. These fragments, termed Enzyme Acceptor (EA) and Enzyme Donor (ED) spontaneously re-associate to form a fully active enzyme that, in the assay format, cleaves a substrate, generating a color change that can be measured spectrophotometrically.
G. Summary of Supporting Data
1. Analytical Performance:
Performance is evaluated at the manufacturer's site on the Beckman Coulter AU680 clinical analyzer.
a) Precision
Precision studies were performed in accordance with CLSI Guideline EP05-A3. Samples were prepared by spiking Oxazepam into drug free urine at the cutoff, 25%, 50%, 75% and 100% above and below the cutoff and tested in both qualitative and semi-quantitative modes. Results presented below were generated by testing all samples in replicates of 2, twice per day for 20 days, total n=80. The results for both cutoffs are summarized in the tables below.
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| Total Precision (n=80) | ||||
|---|---|---|---|---|
| SpikedConcentration(ng/mL) | % of Cutoff(200 ng/mL) | # ofDeterminants | QualitativeImmunoassayResults(Negative/Positive) | Semi-quantitativeImmunoassayResults(Negative/Positive) |
| 0 | -100% | 80 | 80/0 | 80/0 |
| 50 | -75% | 80 | 80/0 | 80/0 |
| 100 | -50% | 80 | 80/0 | 80/0 |
| 150 | -25% | 80 | 80/0 | 79/1 |
| 200 | 100% | 80 | 6/74 | 1/79 |
| 250 | +25% | 80 | 0/80 | 0/80 |
| 300 | +50% | 80 | 0/80 | 0/80 |
| 350 | +75% | 80 | 0/80 | 0/80 |
| 400 | +100% | 80 | 0/80 | 0/80 |
Qualitative and Semi-Quantitative Analysis
b) Spike Recovery
The study was performed using 20 replicates. This study was carried out by testing spiked samples containing Oxazepam at the cutoff calibrator and control levels. The spiked samples were prepared by spiking Oxazepam into drug free urine. Samples were tested in both Qualitative and Semi-Quantitative mode. The qualitative results for both cutoffs are summarized in the tables below.
| Replicates | 150 ng/mL(n=20) | 250 ng/mL(n=20) |
|---|---|---|
| 1 | Negative | Positive |
| 2 | Negative | Positive |
| 3 | Negative | Positive |
| 4 | Negative | Positive |
| 5 | Negative | Positive |
| 6 | Negative | Positive |
| 7 | Negative | Positive |
| 8 | Negative | Positive |
| 9 | Negative | Positive |
| 10 | Negative | Positive |
| 11 | Negative | Positive |
| 12 | Negative | Positive |
| 13 | Negative | Positive |
| 14 | Negative | Positive |
| 15 | Negative | Positive |
Qualitative Data
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| Replicates | 150 ng/mL(n=20) | 250 ng/mL(n=20) |
|---|---|---|
| 16 | Negative | Positive |
| 17 | Negative | Positive |
| 18 | Negative | Positive |
| 19 | Negative | Positive |
| 20 | Negative | Positive |
| Overlap | No | No |
| Relative to C/O | All 20 below C/O | All 20 above C/O |
c) Analytical Recoverv and Linearity
Linearity studies were performed in accordance with CLSI Guideline EP06-A. To demonstrate the dilution linearity for purposes of sample dilution and quality control upto 800 ng/mL assay range, drug free urine was spiked to 900 ng/mL level calibrator using Oxazepam and diluted with drug free urine to generate 8 intermediate levels.
Each sample was run in replicates of five in semi-quantitative mode and the average was used to determine percent recovery compared to the expected target value. The percent recovery is summarized in the table below.
| Level | ExpectedConcentration(ng/mL) | ObservedConcentration(ng/mL) | Average Recovery(%) | Range of Recovery(%) |
|---|---|---|---|---|
| 1 | 0 | 0 | N/A | |
| 2 | 100 | 107.8 | 107.8 | |
| 3 | 200 | 205.8 | 102.9 | |
| 4 | 300 | 289.4 | 96.5 | |
| 5 | 400 | 412.4 | 103.1 | 95.2 – 107.8 |
| 6 | 500 | 517.2 | 103.4 | |
| 7 | 600 | 595.0 | 99.2 | |
| 8 | 700 | 666.2 | 95.2 | |
| 9 | 800 | 766.2 | 95.8 |
d) Method Comparison and Accuracy
The method comparison study was performed in accordance with CLSI Guideline EP09-A3.
One hundred and twenty eight samples were treated with ß-glucuronidase reagent prior to analysis by the CEDIA™ Benzodiazepine Assay in both qualitative and semi-quantitative modes. The results were compared to LC-MS/MS where samples were also treated with ßglucuronidase. The overall concordance between LC-MS/MS and CEDIA™ Benzodiazepine Assay is 97% in Qualitative mode and 96% in Semi-Quantitative mode for high sensitivity 200 ng/mL cutoff.
The qualitative and semi-quantitative results is summarized in the tables below.
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Qualitative Mode Accuracy study with LC-MS/MS as reference method for high sensitivity 200 ng/mL cutoff
| CandidateDeviceResults | < 50% of Cutoffconcentration byLC-MS/MS(< 100 ng/mL) | Near CutoffNegative (Between50% below thecutoff and thecutoff concentrationas determined byLC-MS/MS)(100-199 ng/mL) | Near Cutoff Positive(Between the cutoffand 50% above thecutoff concentrationas determined byLC-MS/MS)(200-300 ng/mL) | High Positives(Greater than50% above cutoffconcentration(> 300 ng/mL) |
|---|---|---|---|---|
| Positive | 0 | 4*b | 13 | 55 |
| Negative | 54 | 2 | 0 | 0 |
Agreement among Positives: 68/68 = 100%
Agreement among Negative: 56/60 = 93%
Semi-Quantitative Mode Accuracy study with LC-MS/MS as reference method for high sensitivity 200 ng/mL cutoff
| CandidateDeviceResults | < 50% of Cutoffconcentration byLC-MS/MS (<100 ng/mL) | Near CutoffNegative (Between50% below thecutoff and thecutoff concentrationas determined byLC-MS/MS)(100-199 ng/mL) | Near CutoffPositive (Betweenthe cutoff and50% above thecutoffconcentration asdetermined byLC-MS/MS)(200-300 ng/mL) | High Positives(Greater than 50%above cutoffconcentration(> 300 ng/mL) |
|---|---|---|---|---|
| Positive | 0 | 4*b | 13 | 55 |
| Negative | 54 | 2 | 0 | 0 |
Agreement among Positives: 67/68 = 99%
Agreement among Negative: 56/60 = 93%
*0 Discordant sample results for high sensitivity 200 ng/mL cutoff
| Qualitative | Semi-Quantitative | LC-MS/MS | |||
|---|---|---|---|---|---|
| Sample ID | Negative/Positive | ObservedConcentration(ng/mL) | Negative/Positive | TotalBenzodiazepineParent Only(ng/mL) | Negative/Positive |
| CA160606-045*1 | Positive | 3168 | Positive | 111 | Negative |
| CA170605-001*1 | Positive | 3723 | Positive | 171 | Negative |
| CA160926-057*1 | Positive | 1441 | Positive | 199 | Negative |
| CA180820-014*2 | Positive | 291 | Positive | 197 | Negative |
*1 These samples are discordant due to the presence of parent benzodiazepine and also benzodiazepine metabolites as follows: CA160606-045 contains 7-aminoclonazepam at 3155 ng/ml. CA170605-001 7-aminoclonazepam at 560 ng/mL. CA160926-057 contains 7-aminoclonazepam at 411 ng/mL and 13 ng/mL of a hydroxyprazolam. *2 Sample CA180820-014 is borderline negative by LC-MS/MS at 197ng/ml compared to the 200 ng/ml cut-off.
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e) Specificity
The cross-reactivity of benzodiazepine compounds were evaluated by adding known amounts of each compound to drug-free negative urine. The results are summarized in the tables below.
| Benzodiazepines and metabolites | Tested Concentration(ng/mL) | Cross-reactivity (%) |
|---|---|---|
| α-Hydroxyalprazolam | 110 | 182 |
| α-Hydroxytriazolam | 140 | 143 |
| Alprazolam | 100 | 200 |
| 7-Aminoclonazepam | 800 | 25 |
| 7-Aminoflunitrazepam | 225 | 89 |
| 7-Aminonitrazepam | 500 | 40 |
| Bromazepam | 300 | 67 |
| Chlordiazepoxide | 2000 | 10 |
| Clobazam | 450 | 44 |
| Clonazepam | 350 | 57 |
| Clorazepate | 100 | 200 |
| Delorazepam | 100 | 200 |
| Demoxepam | 1500 | 13 |
| Desalkylflurazepam(Norfludiazepam) | 110 | 182 |
| Diazepam | 80 | 250 |
| Estazolam | 115 | 174 |
| Flunitrazepam | 125 | 160 |
| Flurazepam | 70 | 286 |
| Lorazepam | 250 | 80 |
| Lorazepam glucuronide | 400 | 50 |
| Lormetazepam | 175 | 114 |
| Medazepam | 200 | 100 |
| Nitrazepam | 290 | 69 |
| Nordiazepam(Desmethyldiazepam) | 70 | 286 |
| Oxazepam | 200 | 100 |
| Oxazepam glucuronide | 350 | 57 |
| Prazepam | 140 | 143 |
| Temazepam | 130 | 154 |
| Temazepam glucuronide | 250 | 80 |
| Triazolam | 90 | 222 |
Cross reactivity of benzodiazepines and metabolites
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Structurally unrelated compounds were evaluated by adding each substance to Oxazepam spiked at 150 ng/mL (-25% of the 200 ng/mL cutoff concentration), 250 ng/mL (+25% of the 200 ng/mL cutoff concentration), at the concentrations indicated. As shown in the table below, the Controls were detected accurately, Low Control as Negative and the High Control as Positive, indicating that all the compounds evaluated exhibited no significant cross-reactivity at the concentrations tested.
| Structurally Unrelated Compounds | TestedConcentration(ng/mL) | Low Control(150 ng/mL) | High Control(250 ng/mL) |
|---|---|---|---|
| 6-Acetyl Morphine | 100,000 | Negative | Positive |
| 10,11 Dihydrocarbamazepine | 100,000 | Negative | Positive |
| 11-nor-Δ9-THC-COOH | 100,000 | Negative | Positive |
| Acetaminophen | 100,000 | Negative | Positive |
| Acetylsalicylic Acid | 100,000 | Negative | Positive |
| Amitriptyline | 75,000 | Negative | Positive |
| Amoxicillin | 100,000 | Negative | Positive |
| Amphetamine | 100,000 | Negative | Positive |
| Benzoylecgonine | 100,000 | Negative | Positive |
| Brompheniramine | 100,000 | Negative | Positive |
| Buprenorphine | 100,000 | Negative | Positive |
| Caffeine | 100,000 | Negative | Positive |
| Captopril | 100,000 | Negative | Positive |
| Cimetidine | 100,000 | Negative | Positive |
| Codeine | 100,000 | Negative | Positive |
| Desipramine | 100,000 | Negative | Positive |
| Dextromethorphan | 100,000 | Negative | Positive |
| Digoxin | 100,000 | Negative | Positive |
| Diphenhydramine | 30,000 | Negative | Positive |
| Enalapril | 100,000 | Negative | Positive |
| EDDP | 100,000 | Negative | Positive |
| EMDP | 3,000 | Negative | Positive |
| Fentanyl | 100,000 | Negative | Positive |
| Fluoxetine | 75,000 | Negative | Positive |
| Fluphenazine | 75,000 | Negative | Positive |
| Haloperidol | 100,000 | Negative | Positive |
| Heroin | 100,000 | Negative | Positive |
| Hydrocodone | 100,000 | Negative | Positive |
| Hydromorphone | 100,000 | Negative | Positive |
| Ibuprofen | 100,000 | Negative | Positive |
| Structurally Unrelated Compounds | TestedConcentration(ng/mL) | Low Control(150 ng/mL) | High Control(250 ng/mL) |
| Levorphanol | 100,000 | Negative | Positive |
| Levothyroxine | 75,000 | Negative | Positive |
| Meperidine | 100,000 | Negative | Positive |
| Methadone | 75,000 | Negative | Positive |
| Methamphetamine | 100,000 | Negative | Positive |
| Morphine | 100,000 | Negative | Positive |
| Morphine-3β-D-glucuronide | 100,000 | Negative | Positive |
| Morphine-6β-D-glucuronide | 100,000 | Negative | Positive |
| Nalbuphine | 100,000 | Negative | Positive |
| Nalorphine | 100,000 | Negative | Positive |
| Naloxone | 100,000 | Negative | Positive |
| Naltrexone | 100,000 | Negative | Positive |
| Naproxen | 100,000 | Negative | Positive |
| Nifedipine | 100,000 | Negative | Positive |
| Oxaprozin | 5,000 | Negative | Positive |
| Oxycodone | 100,000 | Negative | Positive |
| Oxymorphone | 100,000 | Negative | Positive |
| Perphenazine | 30,000 | Negative | Positive |
| Phencyclidine | 90,000 | Negative | Positive |
| Phenobarbital | 100,000 | Negative | Positive |
| Procyclidine | 100,000 | Negative | Positive |
| Propoxyphene | 100,000 | Negative | Positive |
| Ranitidine | 100,000 | Negative | Positive |
| Secobarbital | 100,000 | Negative | Positive |
| Sertraline | 7,000 | Negative | Positive |
| Sulpiride | 100,000 | Negative | Positive |
| Tapentadol | 100,000 | Negative | Positive |
| Thioridazine | 100,000 | Negative | Positive |
| Tramadol | 100,000 | Negative | Positive |
| Triprolidine | 40,000 | Negative | Positive |
| Verapamil | 100,000 | Negative | Positive |
| Zolpidem | 40,000 | Negative | Positive |
| Enalapril | 100,000 | Negative | Positive |
| Salicyluric Acid | 100,000 | Negative | Positive |
| Tolmetin | 100,000 | Negative | Positive |
Structurally unrelated compounds spiked at the concentration listed below into Low and High control urine
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f) Interference
The interference studies were performed in accordance with CLSI Guideline EP07-A2, using both Qualitative and Semi-quantitative modes. The potential interference of pH, endogenous and
{13}------------------------------------------------
exogenous physiologic substances on recovery of Oxazepam using CEDIA™ Benzodiazepine Assay was assessed by spiking known compounds of potentially interfering substances into the Low Control, 150 ng/mL (-25% of the cutoff concentration of 200 ng/mL) and High Control, 250 ng/mL (+25% of the cutoff concentration of 200 ng/mL). In the presence of the compounds listed below, the controls were detected accurately, indicating that these compounds did not show interference in the assay.
| Compound | TestedConcentration(mg/dL) | 200 ng/mL cutoff | |
|---|---|---|---|
| Low Control(150 ng/mL) | High Control(250 ng/mL) | ||
| Acetaminophen | 10 | Negative | Positive |
| Acetone | 500 | Negative | Positive |
| Acetyl Salicylic Acid | 10 | Negative | Positive |
| Ascorbic Acid | 150 | Negative | Positive |
| Caffeine | 5 | Negative | Positive |
| Creatinine | 400 | Negative | Positive |
| Ethanol | 1000 | Negative | Positive |
| Galactose | 5 | Negative | Positive |
| Glucose | 1000 | Negative | Positive |
| Hemoglobin | 150 | Negative | Positive |
| Human Serum Albumin | 200 | Negative | Positive |
| Ibuprofen | 10 | Negative | Positive |
| Oxalic Acid | 50 | Negative | Positive |
| Riboflavin | 3 | Negative | Positive |
| Sodium Chloride | 1000 | Negative | Positive |
| Urea | 1000 | Negative | Positive |
| pH | |||
| pH | 3 | Negative | Positive |
| pH | 4 | Negative | Positive |
| pH | 5 | Negative | Positive |
| pH | 6 | Negative | Positive |
| pH | 7 | Negative | Positive |
| pH | 8 | Negative | Positive |
| pH | 9 | Negative | Positive |
| pH | 10 | Negative | Positive |
| pH | 11 | Negative | Positive |
Interference substances for 200 ng/mL
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g) Specific Gravity
Drug free urine samples with specific gravity ranging in value within 1.002 to 1.029 were split and spiked with Oxazepam to a final concentration of either 150 ng/mL or 225 ng/mL (the Low Control concentrations). These samples were then evaluated in both qualitative and semi-quantitative modes. The Controls were detected accurately, indicating that no interference was observed.
| SpecificGravity | 200 ng/mL cutoff | |
|---|---|---|
| Low Control(150 ng/mL) | High Control(250 ng/mL) | |
| 1.002 | Negative | Positive |
| 1.004 | Negative | Positive |
| 1.005 | Negative | Positive |
| 1.007 | Negative | Positive |
| 1.010 | Negative | Positive |
| 1.012 | Negative | Positive |
| 1.014 | Negative | Positive |
| 1.019 | Negative | Positive |
| 1.023 | Negative | Positive |
| 1.025 | Negative | Positive |
| 1.029 | Negative | Positive |
Specific gravity interference data for 200 ng/mL
H. Conclusion
The information supports a determination of substantial equivalence between CEDIA™ Benzodiazepine Assay and the predicate device CEDIA™ DAU Benzodiazepine Assay (K962734)
§ 862.3170 Benzodiazepine test system.
(a)
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.(b)
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).