(134 days)
Not Found
No
The device description and performance studies focus on a homogeneous enzyme immunoassay and standard analytical methods, with no mention of AI or ML.
No
The device is a diagnostic assay for detecting hydrocodone and its metabolites in urine, not a device used for treating or preventing disease.
Yes
The device is intended for the qualitative and/or semi-quantitative determination of hydrocodone and its metabolites in human urine, providing a preliminary analytical test result for screening, which falls under the definition of a diagnostic device.
No
The device is a homogeneous enzyme immunoassay supplied as liquid reagents, which are physical components, not software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states the device is for the "qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine". This is a diagnostic test performed on a sample taken from the human body.
- Sample Type: The device analyzes "human urine". This is a biological sample.
- Purpose: The assay provides a "preliminary analytical test result" to detect the presence of hydrocodone and its metabolites. This information is used in a laboratory setting to aid in diagnosis or monitoring.
- Device Description: The description details a "homogeneous enzyme immunoassay" that uses specific antibodies to detect substances in the urine sample. This is a common method used in IVD tests.
- Performance Studies: The performance studies describe analytical performance evaluations, method comparison with a confirmatory method (LC-MS/MS), specificity testing, and interference testing. These are all standard evaluations for IVD devices.
- Intended User/Care Setting: The intended user is "laboratories", which is a typical setting for performing IVD tests.
The definition of an In Vitro Diagnostic (IVD) device is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state, or concerning a congenital abnormality, or to monitor therapeutic measures. This device clearly fits this definition.
N/A
Intended Use / Indications for Use
DRI Hydrocodone Assay:
The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine at a cut-off concentration of 300 ng/mL. The assay is intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect hydrocodone and its metabolites in human urine. The assay is designed for use with a number of clinical chemistry analyzers.
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
Product codes (comma separated list FDA assigned to the subject device)
DJG
Device Description
The DRI Hydrocodone assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect Hydrocodone and Hydromorphone and Hydromorphone glucuronide. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in enzyme activity. This phenomenon creates a direct relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.
The assay consists of reagents (A and E).
Reagent A contains mouse monoclonal anti-Hydrocodone antibody, glucose-6-phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as a preservative.
Reagent E: Contains Hydrocodone derivative labeled with glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with sodium azide as a preservative.
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
laboratories
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 was evaluated at the manufacturer's site on an Indiko Plus clinical analyzer.
- Precision: Samples were prepared by spiking Hydrocodone into drug-free urine at the cutoff (100%), 25%, 50%, and 75% above and below the cutoff. These were tested in both qualitative and semi-quantitative modes using a Clinical Laboratory and Standards Institute (CLSI) protocol. Results were generated by testing all samples in replicates of 2, twice per day for 20 days, for a total of n=80 determinations per concentration per lot.
- Key Results (Qualitative mode - Lot #1):
- 0 ng/mL: 80 Negative
- 75 ng/mL: 80 Negative
- 150 ng/mL: 80 Negative
- 225 ng/mL: 80 Negative
- 300 ng/mL: 22 Negative/58 Positive
- 375 ng/mL: 80 Positive
- 450 ng/mL: 80 Positive
- 525 ng/mL: 80 Positive
- 600 ng/mL: 80 Positive
- Key Results (Qualitative mode - Lot #2):
- 0 ng/mL: 80 Negative
- 75 ng/mL: 80 Negative
- 150 ng/mL: 80 Negative
- 225 ng/mL: 80 Negative
- 300 ng/mL: 4 Negative/76 Positive
- 375 ng/mL: 80 Positive
- 450 ng/mL: 80 Positive
- 525 ng/mL: 80 Positive
- 600 ng/mL: 80 Positive
- Key Results (Semi-quantitative mode - Lot #1):
- 0 ng/mL: 80 Negative
- 75 ng/mL: 80 Negative
- 150 ng/mL: 80 Negative
- 225 ng/mL: 80 Negative
- 300 ng/mL: 24 Negative/56 Positive
- 375 ng/mL: 80 Positive
- 450 ng/mL: 80 Positive
- 525 ng/mL: 80 Positive
- 600 ng/mL: 80 Positive
- Key Results (Semi-quantitative mode - Lot #2):
- 0 ng/mL: 80 Negative
- 75 ng/mL: 80 Negative
- 150 ng/mL: 80 Negative
- 225 ng/mL: 80 Negative
- 300 ng/mL: 7 Negative/73 Positive
- 375 ng/mL: 80 Positive
- 450 ng/mL: 80 Positive
- 525 ng/mL: 80 Positive
- 600 ng/mL: 80 Positive
- Key Results (Qualitative mode - Lot #1):
- Spike Recovery: In qualitative mode, there was no ± 2SD overlap between the spiked 300 ng/mL samples and spiked 225 and 375 ng/mL samples. All 20 replicates of spiked 225 ng/mL (Low Control) were below the spiked 300 ng/mL (Cutoff) sample, and all 20 replicates of spiked 375 ng/mL (High Control) samples were above the spiked 300 ng/mL sample. Spike recovery meets Acceptance Criteria.
- Analytical Recovery and Dilution Linearity: Drug free urine spiked to 1000 ng/mL Hydrocodone and diluted to generate 10 intermediate levels. Each sample run in replicates of five in semi-quantitative mode. The percent recovery ranged from 91.1% to 110.3%. The dilution linearity study meets the Acceptance Criteria.
- Method Comparison and Accuracy: One hundred and thirty six patient samples were analyzed by the DRI Hydrocodone Assay in both qualitative and semi-quantitative modes and compared to LC-MS/MS. The overall concordance between LC-MS/MS and DRI Hydrocodone Assay is 82.4%.
- Qualitative Mode Concordance (Sample Sizes within categories not specified, but total 136):
- Positive results: 0 ( 450 ng/mL)
- Negative results: 42 (Negative), 3 ( 450 ng/mL)
- *Discordant samples (7+17) mostly due to cross-reactivity with hydromorphone and hydromorphone-3β-glucuronide.
- Semi-Quantitative Mode Concordance: Same results as qualitative mode for method comparison.
- Qualitative Mode Concordance (Sample Sizes within categories not specified, but total 136):
- Specificity: Cross-reactivity was evaluated.
- Hydrocodone: 100% cross-reactivity at 300 ng/mL.
- Hydromorphone: 92% cross-reactivity at 325 ng/mL.
- Norhydrocodone: 2% cross-reactivity at 13,000 ng/mL.
- Dihydrocodeine: 2% cross-reactivity at 12,500 ng/mL.
- Hydromorphone-3β-glucuronide: 162% cross-reactivity at 185 ng/mL (semi-quantitative mode).
- Minimal cross-reactivity with other opiates and structurally related compounds at tested concentrations.
- No cross-reactivity with structurally unrelated compounds at tested concentrations, as they did not cause false positive or false negative results when spiked with low or high control hydrocodone levels.
- Interference: The potential interference of pH and endogenous physiologic substances (e.g., Acetaminophen, Acetone, Caffeine, Creatinine, Ethanol, Glucose, Hemoglobin, Urea) was assessed by spiking known amounts into Low Control (225 ng/mL) and High Control (375 ng/mL) samples. All tested compounds did not show interference in the assay, with controls detected accurately.
- Specific Gravity: Drug-free urine samples within specific gravity 1.003 to 1.031 were split and spiked to 225ng/mL or 375ng/mL. No interference was observed in qualitative and semi-quantitative modes.
- Traceability: Primary controls and calibrators are traceable to 1 mg/mL Hydrocodone stock solution (99.9% purity from commercial source), confirmed by LC-MS/MS from three independent laboratories.
- Stability:
- Open Vial Stability: Supports 60 days for qualitative and semi-quantitative modes at 2-8°C. (Data from K150502).
- Reagent On-Board Stability: Supports 60 days for qualitative and semi-quantitative modes on clinical analyzer.
- Real Time Stability for Reagent: Low Control negative and High Control positive for 2 years at 2-8°C, with recoveries within 80-120%. Current shelf-life claim is 18 months.
- Detection Limit: Not provided in this type of 510(k) assay.
- Clinical Studies:
- Clinical Sensitivity: Not provided in this type of 510(k) assay.
- Clinical Specificity: Not provided in this type of 510(k) assay.
- Clinical cutoff: Currently no SAMHSA recommendations for hydrocodone clinical cutoff.
- Precision: Samples were prepared by spiking Hydrocodone into drug-free urine at the cutoff (100%), 25%, 50%, and 75% above and below the cutoff. These were tested in both qualitative and semi-quantitative modes using a Clinical Laboratory and Standards Institute (CLSI) protocol. Results were generated by testing all samples in replicates of 2, twice per day for 20 days, for a total of n=80 determinations per concentration per lot.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Overall concordance between LC-MS/MS and DRI Hydrocodone Assay is 82.4%.
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
§ 862.3650 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). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.
February 13, 2018
Microgenics Corporation Minoti Patel Manager, Regulatory Affairs 46500 Kato Road Fremont, CA 94538
Re: K173195
Trade/Device Name: DRI Hydrocodone Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: DJG Dated: January 5, 2018 Received: January 8, 2018
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. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR
1
Part 820); 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Kellie B. Kelm -S
for Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration
Indications for Use
Form Approved: OMB No. 0910-0120 Expiration Date: 06/30/2020 See PRA Statement below.
510(k) Number (if known) K173195
Device Name DRI Hydrocodone Assay
Indications for Use (Describe)
DRI Hydrocodone Assay:
The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine at a cut-off concentration of 300 ng/mL. The assay is intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect hydrocodone and its metabolites in human urine. The assay is designed for use with a number of clinical chemistry analyzers.
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
| 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 NEEDED.
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5. 510(k) Summary - K173195
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, RAC |
| Manager, Regulatory Affairs | |
| Email: Minoti.patel@thermofisher.com | |
| Phone: 510-979-5000 | |
| FAX: 510-979-5002 | |
| Device Common Name: | Homogeneous Hydrocodone Enzyme Immunoassay |
| Trade or Proprietary Name | DRI Hydrocodone Assay |
| Predicate Device Product | |
| Code, Classification, | |
| Classification Name & | |
| Panel | DJG, Class II, 21 CFR 862.3650 – Opiate test system, 91 – |
| Toxicology | |
| Device Classification Name: | 21 CFR Part 862.3650 |
Predicate Device Information:
| Predicate Device: | DRI Hydrocodone Assay |
|---|---|
| Predicate Device | |
| Manufacturer: | Microgenics Corporation |
| Predicate Device Common | |
| Name: | Homogeneous Hydrocodone Enzyme Immunoassay |
| Predicate Device Premarket | |
| Notification #: | K150502 |
| 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 15, 2017
C. Description of Device
The DRI Hydrocodone assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect Hydrocodone and Hydromorphone and Hydromorphone glucuronide. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in
4
enzyme activity. This phenomenon creates a direct relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.
The assay consists of reagents (A and E).
Reagent A contains mouse monoclonal anti-Hydrocodone antibody, glucose-6-phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as a preservative.
Reagent E: Contains Hydrocodone derivative labeled with glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with sodium azide as a preservative.
D. Intended Use
a. Intended Use:
See indications for use below.
b. Indication(s) Use:
DRI Hvdrocodone Assay:
The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine at a cutoff concentration of 300 ng/mL. The assay is intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect hydrocodone and its metabolites in human urine. The assay is designed for use with a number of clinical chemistry analyzers.
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
E. Comparison to Predicate Device
Both devices are exactly same product. The performance data collected for predicate is on AU 680 Analyzer and the performance data collected for this submission is on Indiko Plus. Both devices use same reagent and similar instrument systems. Both devices detect Hydrocodone and its metabolites. Both device's specific antibody detects Hydrocodone and its metabolites at cutoff concentration of 300 ng/mL. Comparison of DRI Hydrocodone Assay, Calibrators, Controls on Indiko and predicate devices demonstrated that the technological characteristics and intended use are substantially equivalent to the currently marketed predicate devices, DRI Hydrocodone Assay, (K150502)
5
F. Test Principle
The DRI Hydrocodone Assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay uses specific antibody that can detect Hydrocodone and its metabolites. The assay is based on competition between a drug labeled with glucose-6phosphate dehydrogenase (G6PDH), and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in enzyme activity. In the presence of free drug, the free drug occupies the antibody binding sites, allowing the drug bound G6PDH to interact with the substrate, resulting in enzyme activity. This phenomenon creates a direct relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 mm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.
G. Summary of Supporting Data
1. Analytical Performance:
Performance is evaluated at the manufacturer's site on Indiko Plus clinical analyzer.
- a) Precision -Samples were prepared by spiking Hydrocodone into drug free urine at the cutoff (100%), 25%, 50% & 75% above and below the cutoff and tested in both qualitative and semi-quantitative modes using a Clinical Laboratory and Standards Institute (CLSI) protocol. Results presented below were generated by testing all samples in replicates of 2, twice per day for 20 days, total n=80.
LC-MS/MS values of spiked samples
Negative urine was spiked in 25% increments or decrements from the cutoff prepared and tested by LC-MS/MS.
| Spiked Conc.
(ng/mL) | LC-MS/MS values
(ng/mL) | % Recovery |
|-------------------------|----------------------------|------------|
| 0 | 0.00 | N/A |
| 75 | 89.50 | 119 |
| 150 | 174.85 | 117 |
| 225 | 242.37 | 108 |
| 300 | 318.50 | 106 |
| 375 | 410.66 | 110 |
| 450 | 501.62 | 111 |
| 525 | 592.11 | 113 |
| 600 | 667.52 | 111 |
The LC-MS/MS results are shown in the table below,
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Qualitative mode
Spiked samples were tested in duplicates twice a day, at least 2 hours apart, for 20 days in qualitative mode. The results of the precision study from 20 days are shown below.
Lot #1
| % of Cutoff | Spiked Conc. (ng/mL) | LC-MS/MS values (ng/mL) | Total Run (n=80)
Number of determinants | Immunoassay Results |
|-------------|----------------------|-------------------------|--------------------------------------------|-----------------------------|
| -100% | 0 | 0.00 | 80 | 80 Negative |
| -75% | 75 | 89.50 | 80 | 80 Negative |
| -50% | 150 | 174.85 | 80 | 80 Negative |
| -25% | 225 | 242.37 | 80 | 80 Negative |
| 100% | 300 | 318.50 | 80 | 22 Negative/
58 Positive |
| +25% | 375 | 410.66 | 80 | 80 Positive |
| +50% | 450 | 501.62 | 80 | 80 Positive |
| +75% | 525 | 592.11 | 80 | 80 Positive |
| +100% | 600 | 667.52 | 80 | 80 Positive |
Lot #2
| % of Cutoff | Spiked Conc. (ng/mL) | LC-MS/MS values (ng/mL) | Total Run (n=80) | |
|---|---|---|---|---|
| Number of determinants | Immunoassay Results | |||
| -100% | 0 | 0.00 | 80 | 80 Negative |
| -75% | 75 | 89.50 | 80 | 80 Negative |
| -50% | 150 | 174.85 | 80 | 80 Negative |
| -25% | 225 | 242.37 | 80 | 80 Negative |
| 100% | 300 | 318.50 | 80 | 4 Negative/ |
| 76 Positive | ||||
| +25% | 375 | 410.66 | 80 | 80 Positive |
| +50% | 450 | 501.62 | 80 | 80 Positive |
| +75% | 525 | 592.11 | 80 | 80 Positive |
| +100% | 600 | 667.52 | 80 | 80 Positive |
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Semi-quantitative mode
Spiked samples were tested in duplicates twice a day, at least 2 hours apart, for 20 days in semi-quantitative mode. The results of the precision study from 20 days are shown below.
| % of Cutoff | Spiked Conc.
(ng/mL) | LC-MS/MS
values (ng/mL) | Total Run (n=80) | |
|-------------|-------------------------|----------------------------|---------------------------|-----------------------------|
| | | | Number of
determinants | Immunoassay
Results |
| -100% | 0 | 0.00 | 80 | 80 Negative |
| -75% | 75 | 89.50 | 80 | 80 Negative |
| -50% | 150 | 174.85 | 80 | 80 Negative |
| -25% | 225 | 242.37 | 80 | 80 Negative |
| 100% | 300 | 318.50 | 80 | 24 Negative/
56 Positive |
| +25% | 375 | 410.66 | 80 | 80 Positive |
| +50% | 450 | 501.62 | 80 | 80 Positive |
| +75% | 525 | 592.11 | 80 | 80 Positive |
| +100% | 600 | 667.52 | 80 | 80 Positive |
Lot #1
Lot #2
| % of Cutoff | Spiked Conc.
(ng/mL) | LC-MS/MS values (ng/mL) | Total Run (n=80) | |
|-------------|-------------------------|-------------------------|------------------------|----------------------------|
| | | | Number of determinants | Immunoassay Results |
| -100% | 0 | 0.00 | 80 | 80 Negative |
| -75% | 75 | 89.50 | 80 | 80 Negative |
| -50% | 150 | 174.85 | 80 | 80 Negative |
| -25% | 225 | 242.37 | 80 | 80 Negative |
| 100% | 300 | 318.50 | 80 | 7 Negative/
73 Positive |
| +25% | 375 | 410.66 | 80 | 80 Positive |
| +50% | 450 | 501.62 | 80 | 80 Positive |
| +75% | 525 | 592.11 | 80 | 80 Positive |
| +100% | 600 | 667.52 | 80 | 80 Positive |
- b) Spike Recovery In qualitative mode, there was no ± 2SD overlap between the spiked 300 ng/mL samples and spiked 225 and 375 ng/mL samples.
All 20 replicates of spiked 225 ng/mL (Low Control) samples were below the spiked 300 ng/mL (Cutoff) sample, and all 20 replicates of spiked 375 ng/mL (High Control) samples were above the spiked 300 ng/mL sample.
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| Replicate | 225 ng/mL
(n=20) | 375 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 |
| 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 below C/O |
Spike Recovery Rate Data
In conclusion, spike recovery meets the Acceptance Criteria.
- 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 of Hydrocodone (1000 ng/mL) and diluted with drug free urine to generate 10 intermediate levels.
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. The percent recovery is summarized in the table below.
| Level | Expected
Concentration
(ng/mL) | Observed
Concentration
(ng/mL) | % Recovery |
|-------|--------------------------------------|--------------------------------------|------------|
| 1 | 0 | 0 | N/A |
| 2 | 100 | 95 | 95.4 |
| 3 | 200 | 219 | 109.3 |
% Recovery of samples
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| Level | Expected
Concentration
(ng/mL) | Observed
Concentration
(ng/mL) | % Recovery |
|-------|--------------------------------------|--------------------------------------|------------|
| 4 | 300 | 331 | 110.3 |
| 5 | 400 | 427 | 106.7 |
| 6 | 500 | 529 | 105.8 |
| 7 | 600 | 624 | 104.1 |
| 8 | 700 | 696 | 99.4 |
| 9 | 800 | 798 | 99.7 |
| 10 | 900 | 893 | 99.2 |
| 11 | 1000 | 911 | 91.1 |
In conclusion, the dilution linearity study meets the Acceptance Criteria.
- d) Method Comparison and Accuracy One hundred and thirty six patient samples are analyzed by the DRI Hydrocodone Assay in both qualitative and semi-quantitative modes and the results are compared to LC-MS/MS. The overall concordance between LC-MS/MS and DRI Hydrocodone Assay is 82.4%.
Qualitative Mode
| Candidate
Device Results | Negative |
450 ng/mL) |
|-----------------------------|----------|------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Positive | 0 | 7* | 17* | 9 | 48 |
| Negative | 42 | 3 | 10 | 0 | 0 |
- Discordant samples
Semi-Quantitative Mode
| Candidate
Device
Results | Negative |
450 ng/mL) |
|--------------------------------|----------|------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Positive | 0 | 7* | 17* | 9 | 48 |
| Negative | 42 | 3 | 10 | 0 | 0 |
- Discordant samples
10
| Previous
| ID | New ID | Immunoassay | LC-MS/MS (ng/mL) | |||
|---|---|---|---|---|---|---|
| Qualitative | ||||||
| mode | Semi | |||||
| quantitative | ||||||
| mode | Hydrocodone | Hydromorphone | Hydromorphone | |||
| 3β-glucuronide | ||||||
| 4 | 46 | Positive | Positive | 126 |