(241 days)
Not Found
No
The device description and performance studies focus on a homogeneous enzyme immunoassay based on recombinant DNA technology and spectrophotometric measurement. There is no mention of AI, ML, or any computational algorithms that would suggest their use in data analysis or interpretation.
No
The CEDIA™ Benzodiazepine Assay is an in vitro diagnostic device used for the qualitative and/or semi-quantitative determination of benzodiazepines in human urine. It is explicitly stated "For In Vitro Diagnostic Use Only" and is used to provide preliminary analytical test results for drug screening, not for treatment.
Yes
The device is explicitly stated as an "In Vitro Diagnostic Use Only" device intended for the determination of benzodiazepines in human urine, which falls under the definition of a diagnostic device.
No
The device is a homogeneous enzyme immunoassay that relies on chemical reagents and a spectrophotometer for measurement, not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Explicit Statement: The document explicitly states "For In Vitro Diagnostic Use Only" in the Intended Use section.
- Intended Use: The assay is intended for the determination of benzodiazepines in human urine, which is a biological specimen. This analysis is performed outside of the body (in vitro).
- Purpose: The purpose is to provide a preliminary analytical test result for the presence of benzodiazepines, which is a diagnostic step.
- Device Description: The description details a laboratory-based assay using chemical reagents and spectrophotometric measurement, typical of IVD devices.
N/A
Intended Use / Indications for 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 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.
Product codes (comma separated list FDA assigned to the subject device)
JXM
Device Description
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.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Trained professionals
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Performance: Performance evaluated on the Beckman Coulter AU680 clinical analyzer.
- Precision: Studies performed in accordance with CLSI Guideline EP05-A3. Samples prepared by spiking Oxazepam into drug-free urine at cutoff, 25%, 50%, 75%, and 100% above and below the cutoff. Tested in qualitative and semi-quantitative modes. Total n=80 samples per spiked concentration (replicates of 2, twice per day for 20 days).
- At 150 ng/mL (-25% of cutoff), 79/80 determined negative in semi-quantitative mode.
- At 200 ng/mL (cutoff), 6/74 (qualitative) and 1/79 (semi-quantitative) determined negative.
- Spike Recovery: Study performed using 20 replicates of spiked samples containing Oxazepam at cutoff calibrator and control levels in drug-free urine. Tested in Qualitative and Semi-Quantitative mode.
- 150 ng/mL (n=20) samples were all negative.
- 250 ng/mL (n=20) samples were all positive.
- Analytical Recover and Linearity: Linearity studies performed in accordance with CLSI Guideline EP06-A. Drug-free urine spiked to 900 ng/mL with Oxazepam and diluted to generate 8 intermediate levels. Each sample run in replicates of five in semi-quantitative mode.
- Average recovery for concentrations from 100 ng/mL to 800 ng/mL ranged from 95.2% to 107.8%.
- Method Comparison and Accuracy: Study performed in accordance with CLSI Guideline EP09-A3. 128 samples treated with ß-glucuronidase and analyzed by CEDIA™ Benzodiazepine Assay (qualitative and semi-quantitative modes). Results compared to LC-MS/MS (also with ß-glucuronidase treatment).
- Overall concordance between LC-MS/MS and CEDIA™ Benzodiazepine Assay: 97% in qualitative mode and 96% in semi-quantitative mode for 200 ng/mL cutoff.
- Qualitative Mode Accuracy (200 ng/mL cutoff):
- Agreement among Positives: 68/68 = 100%
- Agreement among Negative: 56/60 = 93%
- Semi-Quantitative Mode Accuracy (200 ng/mL cutoff):
- Agreement among Positives: 67/68 = 99%
- Agreement among Negative: 56/60 = 93%
- Discordant samples (4 samples where LC-MS/MS was negative and assay was positive) were due to presence of parent benzodiazepine and/or metabolites not fully captured by the LC-MS/MS total benzodiazepine parent-only value.
- Specificity (Cross-reactivity): Evaluated benzodiazepine compounds and structurally unrelated compounds.
- Benzodiazepines and metabolites: Cross-reactivity varies (e.g., α-Hydroxyalprazolam 182%, Alprazolam 200%, 7-Aminoclonazepam 25%, Diazepam 250%, Oxazepam 100%).
- Structurally Unrelated Compounds: 50 compounds (e.g., Acetaminophen, Amphetamine, Caffeine, Morphine) tested at high concentrations (e.g., 100,000 ng/mL). All low controls (150 ng/mL Oxazepam) were negative and all high controls (250 ng/mL Oxazepam) were positive, indicating no significant cross-reactivity.
- Interference: Evaluated potential interference of pH, endogenous, and exogenous physiologic substances (e.g., Acetaminophen, Acetone, Creatinine, Ethanol, Glucose, Hemoglobin, Urea).
- Low Control (150 ng/mL) remained negative and High Control (250 ng/mL) remained positive across tested concentrations and pH ranges (3-11), indicating no interference.
- Specific Gravity: Drug-free urine samples with specific gravity 1.002 to 1.029 spiked with Oxazepam (150 ng/mL or 225 ng/mL).
- Controls detected accurately (low negative, high positive), indicating no interference from specific gravity.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Qualitative Mode Accuracy (200 ng/mL cutoff):
Agreement among Positives: 68/68 = 100%
Agreement among Negative: 56/60 = 93%
Semi-Quantitative Mode Accuracy (200 ng/mL cutoff):
Agreement among Positives: 67/68 = 99%
Agreement among Negative: 56/60 = 93%
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 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).
0
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
1
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
2
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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3
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 Corporation |
| Thermo Fisher Scientific | |
| 46500 Kato Road | |
| Fremont, CA 94538 | |
| Phone: 510-979-5000 | |
| FAX: 510-979-5002 | |
| Correspondent Contact | |
| Information: | Minoti Patel |
| Director, Regulatory Affairs | |
| Email: Minoti.patel@thermofisher.com | |
| Phone: 510-979-5000 | |
| FAX: 510-979-5002 | |
| Device Common Name: | Benzodiazepine Enzyme Immunoassay |
| Trade or Proprietary Name | CEDIATM Benzodiazepine Assay |
| Candidate Device Product | |
| Code, Classification, | |
| Classification Name & | |
| Panel | JXM, Class II, 21 CFR 862. 3170 – Benzodiazepine |
| Test 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.
4
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.
5
E. Comparison to Predicate Device
| Characteristic | Candidate Device:
CEDIATM Benzodiazepine Assay | Predicate Device:
CEDIATM DAU Benzodiazepine
(K962734) |
|-------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------|
| Intended Use | The CEDIATM 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. | Same |
| Operating Principle
(Technology) | CEDIATM | Same |
| Characteristic | Candidate Device:
CEDIA™ Benzodiazepine Assay | Predicate Device:
CEDIA™ DAU Benzodiazepine
(K962734) |
| Measured Analyte | Benzodiazepines and their
metabolites | Same |
| Test Matrix | Urine | Same |
| Cutoff Levels | HS 200 ng/mL | Same |
| Methodology | Homogeneous Enzyme
Immunoassay | 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 |
6
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.
7
| Total Precision (n=80) | ||||
|---|---|---|---|---|
| Spiked | ||||
| Concentration | ||||
| (ng/mL) | % of Cutoff | |||
| (200 ng/mL) | # of | |||
| Determinants | Qualitative | |||
| Immunoassay | ||||
| Results | ||||
| (Negative/Positive) | Semi-quantitative | |||
| Immunoassay | ||||
| Results | ||||
| (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
8
| 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 | Expected
Concentration
(ng/mL) | Observed
Concentration
(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.
9
Qualitative Mode Accuracy study with LC-MS/MS as reference method for high sensitivity 200 ng/mL cutoff
| Candidate
Device
Results | 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
| Candidate
Device
Results | 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 | Observed | ||||
| Concentration | |||||
| (ng/mL) | Negative/ | ||||
| Positive | Total | ||||
| Benzodiazepine | |||||
| Parent 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.
10
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
11
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 | Tested
Concentration
(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 | Tested
Concentration
(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
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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 | Tested
Concentration
(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.
| Specific
| Gravity | 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)