(59 days)
Not Found
No
The summary describes a standard enzyme immunoassay for detecting a specific metabolite in urine. There is no mention of AI, ML, or any computational methods beyond basic analytical calculations for qualitative and semi-quantitative results. The performance studies focus on traditional analytical metrics like precision, recovery, and method comparison, not AI/ML-specific metrics or training/test sets in the context of model development.
No.
This device is an in vitro diagnostic assay used to detect the presence of heroin metabolite in human urine, not to treat a medical condition.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is for "in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine" and provides a "rapid analytical screening procedure to detect 6-Acetylmorphine in human urine." This directly aligns with the definition of a diagnostic device, which is used to diagnose diseases or conditions.
No
The device is an in vitro diagnostic assay consisting of buffers and lyophilized reagents, which are physical components, not software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine..." and "For In Vitro Diagnostic Use Only".
- Nature of the Test: The assay analyzes a sample (human urine) outside of the body to detect a substance (heroin metabolite). This is the fundamental definition of an in vitro diagnostic test.
- Intended Use: The assay is intended to be used in laboratories by trained professionals for analytical screening of human urine. This aligns with the typical use of IVD devices.
N/A
Intended Use / Indications for Use
The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only.
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 with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.
Clinical 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
DJG
Device Description
The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include mouse monoclonal antibodies to 6-Acetylmorphine, recombinant microbial "enzyme donor" - 6-Acetylmorphine conjugate, "enzyme acceptor", chlorophenol red ß-Dgalactopyranoside, stabilizers and preservatives.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
human urine
Indicated Patient Age Range
Not Found
Intended User / Care Setting
laboratories, trained professionals only.
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
- Analytical Performance:
- Precision Study: Performed with two replicates per run, twice a day, for 20 days (total n=80). Samples were prepared by spiking 6-Acetylmorphine methanol stock solution into drug-free urine at the cutoff, 25%, 50%, 75% & 100% above and below the cutoff and tested in both qualitative and semi-quantitative modes, using a Clinical Laboratory and Standards Institute (CLSI) protocol.
- Qualitative Results:
- 0 ng/mL ( -100% of Cutoff): 80 determinations, 80/0 (Negative/Positive)
- 2.5 ng/mL ( -75% of Cutoff): 80 determinations, 80/0
- 5 ng/mL ( -50% of Cutoff): 80 determinations, 80/0
- 7.5 ng/mL ( -25% of Cutoff): 80 determinations, 80/0
- 10 ng/mL (100% of Cutoff): 80 determinations, 25/55
- 12.5 ng/mL ( +25% of Cutoff): 80 determinations, 0/80
- 15 ng/mL ( +50% of Cutoff): 80 determinations, 0/80
- 17.5 ng/mL ( +75% of Cutoff): 80 determinations, 0/80
- 20 ng/mL ( +100% of Cutoff): 80 determinations, 0/80
- Semi-Quantitative Results:
- 0 ng/mL ( -100% of Cutoff): 80 determinations, 80/0
- 2.5 ng/mL ( -75% of Cutoff): 80 determinations, 80/0
- 5 ng/mL ( -50% of Cutoff): 80 determinations, 80/0
- 7.5 ng/mL ( -25% of Cutoff): 80 determinations, 80/0
- 10 ng/mL (100% of Cutoff): 80 determinations, 46/34
- 12.5 ng/mL ( +25% of Cutoff): 80 determinations, 0/80
- 15 ng/mL ( +50% of Cutoff): 80 determinations, 0/80
- 17.5 ng/mL ( +75% of Cutoff): 80 determinations, 0/80
- 20 ng/mL ( +100% of Cutoff): 80 determinations, 0/80
- Qualitative Results:
- Spike Recovery:
- Qualitative: No ± 2SD overlap between 10 ng/mL, 7.5ng/mL, and 12.5 ng/mL samples. All 20 replicates of spiked 7.5 ng/mL and 12.5 ng/mL samples detected as Negative and Positive, respectively, when compared to 10 ng/mL spiked sample.
- Semi-quantitative: Spiked samples recovered within 80-120% of the nominal values.
- Analytical Recovery and Dilution Linearity: Drug-free urine spiked to 20ng/mL with 6-Acetylmorphine and serially diluted to generate 10 intermediate levels. Each sample run in replicates of five in semi-quantitative mode.
- Recovery ranged from 97.3% to 115.6%.
- Method Comparison and Accuracy: One hundred and three patient samples analyzed by the CEDIA Heroin Metabolite (6-AM) Assay (qualitative and semi-quantitative modes) and compared to LC-MS/MS.
- Overall concordance between LC-MS/MS and CEDIA Heroin Metabolite (6-Acetylmorphine) Assay was 100%.
- Specificity: Cross-reactivity of Heroin Metabolite (6-AM) and its metabolites evaluated. 6-Acetylmorphine showed 100% cross-reactivity. Heroin showed 8.3% cross-reactivity.
- Cross-reactivity of Structurally Related or Unrelated Opiate Compounds: Various compounds tested, with most showing 10 ng/mL: 80/80, 80/80, 80/80, 80/80.
- At Cutoff (10 ng/mL): 25 Negative, 55 Positive out of 80 determinations.
- Precision Study: Performed with two replicates per run, twice a day, for 20 days (total n=80). Samples were prepared by spiking 6-Acetylmorphine methanol stock solution into drug-free urine at the cutoff, 25%, 50%, 75% & 100% above and below the cutoff and tested in both qualitative and semi-quantitative modes, using a Clinical Laboratory and Standards Institute (CLSI) protocol.
- Semi-Quantitative Results (Precision Study):
- Negative Agreement for 10 ng/mL: 80/80, 80/80, 80/80, 80/80.
- At Cutoff (10 ng/mL): 46 Negative, 34 Positive out of 80 determinations.
- Method Comparison and Accuracy (Qualitative Results):
- True Negative (LC-MS/MS = 10 ng/mL, Device Positive): 6 + 45 = 51
- False Positive: 0
- False Negative: 0
- Overall Concordance: 100%
- Method Comparison and Accuracy (Semi-Quantitative Results):
- True Negative (LC-MS/MS = 10 ng/mL, Device Positive): 6 + 45 = 51
- False Positive: 0
- False Negative: 0
- Overall Concordance: 100%
- Specificity (Cross-reactivity):
- 6-Acetylmorphine: 100%
- Heroin: 8.3%
- Various other opiates and structurally related compounds show varying, mostly low, cross-reactivity.
- Structurally unrelated compounds showed minimal cross-reactivity.
- Analytical Recovery: Average recovery for semi-quantitative mode ranged from 97.3% to 115.6%.
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 862.3650 Opiate test system.
(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate 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, which is a blue square with the letters "FDA" in white. To the right of the FDA logo is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
December 16, 2019
Microgenics Corporation Sharron Penn Senior Regulatory Affairs Specialist 46500 Kato Road Fremont, CA 94538
Re: K192943
Trade/Device Name: CEDIA Heroin Metabolite (6-AM) Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: DJG Dated: October 17, 2019 Received: October 18, 2019
Dear Sharron Penn:
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.
1
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 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 (QS) 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, M.T., 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) K192943
Device Name CEDIA Heroin Metabolite (6-AM) Assay
Indications for Use (Describe)
The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only.
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 with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.
Clinical 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) |
CONTINUE ON A SEPARATE PAGE IF NEEDEDThis section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
3
1. 510(k) Summary
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: | Sharron Penn |
| Senior Regulatory Affairs Specialist | |
| Email: Sharron.Penn@thermofisher.com | |
| Phone: 510-979-5000 | |
| FAX: 510-979-5002 | |
| Device Common Name: | 6-Acetylmorphine Immunoassay Test |
| Trade or Proprietary Name | CEDIA Heroin Metabolite (6-AM) Assay |
| Predicate Device Product | |
| Code, Classification, | |
| Classification Name & Panel | DJG, Class II, 21 CFR 862.3650 – Opiate test system, |
| 91 - Toxicology |
Predicate Device Information:
| Predicate Device: | CEDIA Heroin Metabolite (6-AM) Assay |
|---|---|
| Predicate Device | |
| Manufacturer: | Microgenics Corporation |
| Predicate Device Common | |
| Name: | 6-Acetylmorphine Immunoassay Test System |
| Predicate Device Premarket | |
| Notification #: | K173183 |
| Predicate Device Product | |
| Code, Classification, | |
| Classification Name & Panel | DJG, Class II, 21 CFR 862.3650 ">– Opiate test system, |
| 91 – Toxicology |
B. Date Summary Prepared December 12, 2019
C. Description of Device
The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include mouse monoclonal antibodies to 6-Acetylmorphine, recombinant microbial "enzyme donor" - 6-Acetylmorphine conjugate, "enzyme acceptor", chlorophenol red ß-Dgalactopyranoside, stabilizers and preservatives.
4
D. Intended Use
CEDIA Heroin Metabolite (6-AM) Assay
The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only.
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 with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.1
Clinical 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.
| Characteristic | Predicate Device: CEDIA Heroin Metabolite
(6-AM) Assay. K173183. | CEDIA Heroin
Metabolite (6-AM)
Assay |
|----------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------|
| Intended Use | The CEDIA Heroin Metabolite (6-AM) Assay is a
homogeneous enzyme immunoassay for the in vitro
qualitative and/or semi-quantitative determination
of the presence of heroin metabolite (6-AM) in
human urine at a cut-off concentration of 10 ng/mL.
The assay is intended to be used in laboratories and
provides a rapid analytical screening procedure to
detect 6-Acetylmorphine in human urine. The assay
is designed for use with a number of clinical
chemistry analyzers. This product is intended to be
used by trained professionals only.
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. | same |
E. Comparison to Predicate Device
5
| Characteristic | Predicate Device: CEDIA Heroin Metabolite
(6-AM) Assay. K173183. | CEDIA Heroin
Metabolite (6-AM)
Assay |
|-------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------|
| | 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
with tandem mass spectrometry (LC-MS/MS) is
the preferred confirmatory method
Clinical 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. | |
| Operating Principle
(Technology) | CEDIA | Same |
| Measured Analyte | Heroin and 6-Acetylmorphine | Same |
| Test Matrix | Urine | Same |
| Cutoff Levels | 10 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 | Mouse Monoclonal antibody | Same |
| Storage | 2-8°C until expiration date. | Same |
| Principal Operator | Trained professionals | Same |
| Reference Instrument | Beckman AU680 | Horiba Pentra C400 |
F. Test Principle
The CEDIA Heroin Metabolite Assay uses recombinant DNA technology (US Patent No. 4708929) to produce a unique homogeneous enzyme immunoassay system.10 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:
- a) Precision Study is performed for two replicates per run, twice a day, for 20 days (total n=80). Samples are prepared by spiking 6-Acetylmorphine methanol stock solution into drug free urine at the cutoff, 25%, 50%, 75% & 100% above and below the cutoff and tested in both qualitative and semi-quantitative modes, using a Clinical Laboratory and Standards Institute (CLSI) protocol.
6
Qualitative Results:
| 6-
Acetylmorphine | % of Cutoff
(10 ng/mL) | LC-MS/MS
(ng/mL) | Total Precision (n=80) | |
|------------------------------------|---------------------------|---------------------|-----------------------------|-----------------------------------------------|
| Spiked
Concentration
(ng/mL) | | | Number of
Determinations | Immunoassay
Results
(Negative/Positive) |
| 0 | -100 | N/A | 80 | 80/0 |
| 2.5 | -75 | 2.6 | 80 | 80/0 |
| 5 | -50 | 5.1 | 80 | 80/0 |
| 7.5 | -25 | 7.6 | 80 | 80/0 |
| 10 | 100 | 10.0 | 80 | 25/55 |
| 12.5 | +25 | 12.6 | 80 | 0/80 |
| 15 | +50 | 15.0 | 80 | 0/80 |
| 17.5 | +75 | 17.6 | 80 | 0/80 |
| 20 | +100 | 20.0 | 80 | 0/80 |
Semi-Quantitative Results:
| 6-
Acetylmorphine
Spiked
Concentration
(ng/mL) | % of Cutoff
(10 ng/mL) | LC-MS/MS
(ng/mL) | Total Precision (n=80) | |
|------------------------------------------------------------|---------------------------|---------------------|-----------------------------|-----------------------------------------------|
| | | | Number of
Determinations | Immunoassay
Results
(Negative/Positive) |
| 0 | -100 | N/A | 80 | 80/0 |
| 2.5 | -75 | 2.6 | 80 | 80/0 |
| 5 | -50 | 5.1 | 80 | 80/0 |
| 7.5 | -25 | 7.6 | 80 | 80/0 |
| 10 | 100 | 10.0 | 80 | 46/34 |
| 12.5 | +25 | 12.6 | 80 | 0/80 |
| 15 | +50 | 15.0 | 80 | 0/80 |
| 17.5 | +75 | 17.6 | 80 | 0/80 |
| 20 | +100 | 20.0 | 80 | 0/80 |
- b) Spike Recovery In qualitative mode, there is no ± 2SD overlap between the spiked 10 ng/mL, 7.5ng/mL and 12.5 ng/mL samples.
All 20 replicates of spiked 7.5 ng/mL and 12.5 ng/mL samples detect as Negative and Positive, respectively, when compared to 10 ng/mL spiked sample.
In semi-quantitative mode, the spiked samples recover within 80-120% of the nominal values.
- c) Analytical Recovery and Dilution Linearity -To demonstrate the dilution linearity for purpose of sample dilution and quality control of the entire assay range, drug free urine is spiked to the high calibrator level (20ng/mL) by using 6-Acetylmorphine methanol solution and diluted with drug free urine to generate 10 intermediate levels.
7
Each sample is run in replicates of five in semi-quantitative mode and the average is used to determine percent recovery compared to the expected target value.
| Level | Target
Concentration
(ng/mL) | Observed
Concentration
(ng/mL) | Average
Recovery (%) | Range of
Recovery (%) |
|-------|------------------------------------|--------------------------------------|-------------------------|--------------------------|
| 1 | 0 | 0.39 | N/A | N/A |
| 2 | 2 | 2.31 | 115.6 | 92.0 - 134.0 |
| 3 | 4 | 4.62 | 115.6 | 100.8 - 130.5 |
| 4 | 6 | 6.25 | 104.2 | 97.7 - 113.5 |
| 5 | 8 | 8.04 | 100.5 | 94.6 – 106.5 |
| 6 | 10 | 10.19 | 101.9 | 97.0 – 108.4 |
| 7 | 12 | 12.33 | 102.7 | 99.6 – 105.1 |
| 8 | 14 | 14.36 | 102.6 | 98.3 – 106.1 |
| 9 | 16 | 15.81 | 98.8 | 95.4 - 100.1 |
| 10 | 18 | 18.27 | 101.5 | 98.1 - 103.3 |
| 11 | 20 | 19.46 | 97.3 | 95.5 - 99.0 |
- d) Method Comparison and Accuracy One hundred and three patient samples were analyzed by the CEDIA Heroin Metabolite (6-AM) Assay in both qualitative and semiquantitative modes and the results were compared to LC-MS/MS. The overall concordance between LC-MS/MS and CEDIA Heroin Metabolite (6-Acetylmorphine) Assay was 100% for the samples evaluated in this study.
Qualitative Results
| Candidate
Device
Results | Negative | 15.0
ng/mL) |
|--------------------------------|----------|------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |
8
Semi-Quantitative Results
| Candidate
Device
Results | Negative | 15.0
ng/mL) |
|--------------------------------|----------|------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |
- e) Specificity The cross-reactivity of Heroin Metabolite (6-AM) and its metabolites was evaluated by adding known amounts of each analyte to drug-free negative urine. As indicated by the results in the table below, 6-Acetylmorphine showed 100% crossreactivity. Heroin showed 8.3% cross-reactivity.
| Heroin Metabolite (6-AM) and its
metabolites | Tested Concentration
(ng/mL) | Positive/
Negative | Cross-
reactivity
(%) |
|-------------------------------------------------|---------------------------------|-----------------------|-----------------------------|
| 6-Acetylmorphine | 10 | Positive | 100 |
| Heroin | 120 | Positive | 8.3 |
Cross Reactivity of Structurally Related or Unrelated Opiate Compounds
| Structurally related compounds and
other opiates | Tested
Concentration
(ng/mL) | Positive/
Negative | Cross-reactivity (%) |
|-----------------------------------------------------|------------------------------------|-----------------------|----------------------|
| 6-Acetylcodeine | 100,000 | Negative |