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

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.

AI/ML Overview

The provided document is a 510(k) premarket notification for a medical device called the DRI Hydrocodone Assay. This device is a homogeneous enzyme immunoassay designed for the qualitative and/or semi-quantitative determination of hydrocodone and its metabolites in human urine.

Here's an analysis of the acceptance criteria and study proving the device meets those criteria, based on the provided text:

Key Takeaway: This 510(k) is for a new instrument platform (Indiko Plus) for an already cleared assay (DRI Hydrocodone Assay, K150502). Therefore, much of the study focuses on showing comparable performance between the new instrument and the previous one. The acceptance criteria are implicitly met by demonstrating performance consistent with an already legally marketed predicate device.

1. Table of Acceptance Criteria and Reported Device Performance

Since this is a 510(k) for an existing assay on a new instrument, the "acceptance criteria" are not explicitly stated as quantitative targets (e.g., "sensitivity must be >95%"). Instead, the acceptance criteria are implicitly that the performance on the new analyzer (Indiko Plus) is comparable or equivalent to the performance on the predicate analyzer (AU 680). The study aims to demonstrate that the new device performs as intended and similarly to its predicate.

Test Category	Acceptance Criteria (Implicit)	Reported Device Performance (DRI Hydrocodone Assay on Indiko Plus)
Precision	Consistent and reliable results across replicates and days, reflecting expected performance relative to the cutoff. For qualitative mode, no overlap in 2SD ranges between critical concentrations (e.g., -25% and 100% of cutoff, and 100% and +25% of cutoff).	Qualitative Mode:- Lot #1: - -100% to -25% of cutoff: 80/80 Negative results. - 100% of cutoff (300 ng/mL, LC-MS/MS 318.50): 22 Negative / 58 Positive. - +25% to +100% of cutoff: 80/80 Positive results.- Lot #2: - -100% to -25% of cutoff: 80/80 Negative results. - 100% of cutoff (300 ng/mL, LC-MS/MS 318.50): 4 Negative / 76 Positive. - +25% to +100% of cutoff: 80/80 Positive results.Semi-quantitative Mode:- Lot #1: - -100% to -25% of cutoff: 80/80 Negative results. - 100% of cutoff (300 ng/mL, LC-MS/MS 318.50): 24 Negative / 56 Positive. - +25% to +100% of cutoff: 80/80 Positive results.- Lot #2: - -100% to -25% of cutoff: 80/80 Negative results. - 100% of cutoff (300 ng/mL, LC-MS/MS 318.50): 7 Negative / 73 Positive. - +25% to +100% of cutoff: 80/80 Positive results.Spike Recovery: No ± 2SD overlap between 300 ng/mL samples and 225 ng/mL (all 20 replicates negative) and 375 ng/mL (all 20 replicates positive). Conclusion: Meets Acceptance Criteria.
Analytical Recovery and Dilution Linearity	Expected: % Recovery should be within an acceptable range (e.g., 80-120%). Dilution should demonstrate linearity.	Samples run in replicates of five in semi-quantitative mode. % Recovery ranged from 91.1% (at 1000 ng/mL) to 110.3% (at 300 ng/mL for one lot, 200 ng/mL for another). Conclusion: Meets Acceptance Criteria.
Method Comparison and Accuracy	High overall concordance with the confirmatory method (LC-MS/MS). Discordant samples should be explainable.	Overall concordance with LC-MS/MS: 82.4%.- Qualitative & Semi-Quantitative Mode (combined table in document): - True Negatives: 42 - False Positives (immunoassay positive, LC-MS/MS <150 ng/mL or 150-299.9 ng/mL): 7+17 = 24 discordant samples (e.g., Immmunoassay positive, LC-MS/MS 126 ng/mL, 179 ng/mL hydromorphone-3β-glucuronide). - True Positives (immunoassay positive, LC-MS/MS >=300 ng/mL): 9+48 = 57 - False Negatives (immunoassay negative, LC-MS/MS <150 ng/mL or 150-299.9 ng/mL): 3+10 = 13 - False Negatives (immunoassay negative, LC-MS/MS >=300 ng/mL): 0Explanation for discordant samples: Cross-reactivity of immunoassay to hydromorphone and hydromorphone-3β-glucuronide.
Specificity (Cross-reactivity)	Known related compounds should show expected cross-reactivity. Structurally unrelated compounds should show minimal or no cross-reactivity, especially at critical concentrations.	Hydrocodone & Metabolites:- Hydrocodone: 100% cross-reactivity at 300 ng/mL (cutoff).- Hydromorphone: 92% cross-reactivity at 325 ng/mL.- Hydromorphone-3β-glucuronide: 162% cross-reactivity at 185 ng/mL (higher than 100% suggests higher potency than Hydrocodone in the assay).- Norhydrocodone: 2% at 13,000 ng/mL.- Dihydrocodeine: 2% at 12,500 ng/mL.Opiates & Structurally Related: Most showed <0.1% to <0.4% cross-reactivity at high concentrations (e.g., Codeine <0.2% at 150,000 ng/mL), indicating minimal/negligible assay interference from standard opiates at typical physiological levels relative to hydrocodone. Some showed higher e.g., Levorphanol (1.4% at 22,000 ng/mL), Naloxone (1.8% at 17,000 ng/mL), Naltrexone (0.4% at 75,000 ng/mL), Noroxycodone (0.3% at 110,000 ng/mL), Oxycodone (2.1% at 14,000 ng/mL), Oxymorphone-beta-D-glucuronide (2.1% at 14,000 ng/mL), Oxymorphone (2.1% at 140,000 ng/mL).Structurally Unrelated Compounds: All tested compounds (e.g., Acetaminophen, Ibuprofen, Amphetamine, Caffeine) showed no interference; samples spiked with Low Control (225 ng/mL hydrocodone) remained negative, and those with High Control (375 ng/mL hydrocodone) remained positive.
Interference (pH, Endogenous Substances)	No significant interference from common urinary pH variations or endogenous substances at physiologically relevant concentrations.	Interference Substances: No interference observed for common substances (e.g., Acetaminophen, Creatinine, Glucose, Hemoglobin, Urea) at specified concentrations. Low Control remained Negative, High Control remained Positive.pH: No interference observed across pH 4-10. Low Control remained Negative, High Control remained Positive.
Specific Gravity	No significant interference from varying specific gravity of urine samples.	No interference observed for specific gravity ranging from 1.003 to 1.031. Low Control remained Negative, High Control remained Positive.
Stability	Assay reagents should maintain performance over claimed shelf-life and on-board stability periods.	Open Vial Stability: 60 days (validated in predicate submission K150502).Reagent On-Board Stability: 60 days.Real Time Stability: 2 years at 2-8°C, with Low Control remaining negative and High Control remaining positive, and recoveries within 80-120%. Current shelf-life claim is 18 months, which is supported.
Traceability	Calibrators and controls should be traceable to a reliable reference standard.	Primary controls and calibrators are traceable to 1 mg/mL Hydrocodone stock solution (99.9% purity) from a commercial source. Confirmed by LC-MS/MS from three independent laboratories.

2. Sample Size Used for the Test Set and Data Provenance

Precision:
- Qualitative & Semi-quantitative Modes: Samples prepared by spiking hydrocodone into drug-free urine at cutoff, ±25%, ±50%, ±75%, and -100% (0 ng/mL hydrocodone).
- Sample Size: For each of the two reagent lots, each concentration was tested in replicates of 2, twice per day for 20 days.
  - Total n = 80 determinations per concentration per lot.
Analytical Recovery and Dilution Linearity:
- Sample Size: 10 intermediate dilution levels generated from a high calibrator (1000 ng/mL) spiked in drug-free urine. Each sample run in replicates of five. (Therefore, 11 concentrations total * 5 replicates = 55 measurements per lot, though not explicitly stated if multiple lots were used here).
Method Comparison and Accuracy:
- Sample Size: One hundred and thirty six (136) patient samples.
- Data Provenance: Not explicitly stated (e.g., country of origin), but implied to be from a laboratory setting. The terms "patient samples" suggest real-world samples, but it's not specified if they were retrospectively collected or prospectively collected. Given that it's compared against LC-MS/MS, it's likely retrospective.
Specificity (Cross-reactivity) & Interference & Specific Gravity:
- Sample Size:
  - Hydrocodone and its Metabolites: Tested using varying concentrations of each compound, with results given as concentrations achieving cutoff-equivalent response. Number of replicates per concentration not explicitly stated but implied multiple for curve fitting.
  - Opiates and Structurally Related Compounds: Tested at single high concentrations.
  - Structurally Unrelated Compounds: Spiked into 225 ng/mL or 375 ng/mL hydrocodone urine. Tested in 5 replicates for each compound and concentration.
  - Interference Substances & pH & Specific Gravity: Tested at single concentrations/pH values. Replicates not explicitly stated for these specific tests but implied by general "analytical performance" section to follow CLSI protocol, which often involves replicates.
- Data Provenance: Controlled laboratory experiments using spiked drug-free urine.

3. Number of Experts Used to Establish Ground Truth for Test Set and Qualifications

Ground Truth Method: LC-MS/MS (Liquid Chromatography/Tandem Mass Spectrometry) is used as the preferred confirmatory method for hydrocodone and its metabolites.
Experts: LC-MS/MS is a highly precise and definitive analytical chemical method, considered the gold standard for drug quantification in urine. Therefore, the "ground truth" is established by the analytical method itself, not by human experts interpreting results. The document notes that external laboratories confirmed the concentration of primary control and calibrator stocks by LC-MS/MS, adding a layer of external validation to the method's accuracy.

4. Adjudication Method for the Test Set

Adjudication: Not applicable. The "ground truth" is established by an objective, quantitative laboratory assay (LC-MS/MS), not by human interpretation requiring adjudication. Discordant results are analyzed against this LC-MS/MS truth and explained (e.g., cross-reactivity).

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

MRMC Study: No. This device is an immunoassay for drug detection, where the output is a chemical reaction read by an analyzer to provide a qualitative or semi-quantitative result. It does not involve human readers interpreting images or complex data in the same way an AI for radiology would. Therefore, an MRMC study is not relevant or applicable.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Standalone Performance: Yes, in essence. The studies for precision, analytical recovery, specificity, and interference evaluate the assay's performance independent of human interpretation for the final qualitative/quantitative determination. The device's output (positive/negative, or semi-quantitative concentration) is directly compared to the LC-MS/MS ground truth. Human involvement is limited to sample preparation, loading the instrument, and reviewing automated results, not interpreting raw data or making diagnostic decisions without the device's output.

7. The Type of Ground Truth Used

Type of Ground Truth: The ground truth for this device's performance studies is definitive analytical chemistry data, specifically Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) results. This is considered the gold standard for confirming and quantifying drug concentrations in biological samples. It's a highly objective and quantitative method.

8. The Sample Size for the Training Set

Training Set: Not explicitly mentioned or applicable in the context of this traditional immunoassay. This is not a machine learning/AI device that requires a distinct "training set" in the computational sense. Immunoassays leverage established biochemical principles and reagents. The "training" for such devices involves reagent formulation, calibration curve development, and optimization by the manufacturer, rather than machine learning on a dataset. The precision study uses two lots of reagents, which implies a level of manufacturing consistency, but these are not 'training sets' in the AI sense.

9. How the Ground Truth for the Training Set Was Established

Ground Truth for Training/Development: Not applicable as there is no computational "training set" in the AI sense. The development and optimization of the immunoassay reagents rely on known chemical properties, antibody specificity, and the relationship between drug concentration and enzyme activity. Calibrators and controls are used during development and routine use, and their concentrations are traceable to LC-MS/MS confirmed stock solutions (as noted in section 1g, "Traceability"). This analytical method ensures the accuracy of the calibrators used to establish the assay's performance curve.

Summary

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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

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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

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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 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.

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.

This 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."

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5. 510(k) Summary - K173195

A. Device Information

Category	Comments
Sponsor:	Microgenics CorporationThermo Fisher Scientific46500 Kato RoadFremont, CA 94538Phone: 510-979-5000FAX: 510-979-5002
Correspondent ContactInformation:	Minoti Patel, RACManager, Regulatory AffairsEmail: Minoti.patel@thermofisher.comPhone: 510-979-5000FAX: 510-979-5002
Device Common Name:	Homogeneous Hydrocodone Enzyme Immunoassay
Trade or Proprietary Name	DRI Hydrocodone Assay
Predicate Device ProductCode, 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 DeviceManufacturer:	Microgenics Corporation
Predicate Device CommonName:	Homogeneous Hydrocodone Enzyme Immunoassay
Predicate Device PremarketNotification #:	K150502
Predicate Device ProductCode, 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

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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.

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)

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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/MSvalues (ng/mL)	Total Run (n=80)
			Number ofdeterminants	ImmunoassayResults
-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	ExpectedConcentration(ng/mL)	ObservedConcentration(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	ExpectedConcentration(ng/mL)	ObservedConcentration(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

CandidateDevice Results	Negative	< 50% of Cutoffconcentration byLC-MS/MS (<150 ng/mL)	Near CutoffNegative (Between50% below thecutoff and the cutoffconcentration asdetermined by LC-MS/MS) (150 -299.9 ng/mL)	Near Cutoff Positive(Between the cutoffand 50% above thecutoff concentrationas determined by LC-MS/MS) (300 - 450ng/mL)	High Positives(Greater than50% above cutoffconcentration (>450 ng/mL)
Positive	0	7*	17*	9	48
Negative	42	3	10	0	0

Discordant samples

Semi-Quantitative Mode

CandidateDeviceResults	Negative	< 50% of Cutoffconcentration byLC-MS/MS (<150 ng/mL)	Near CutoffNegative (Between50% below thecutoff and the cutoffconcentration asdetermined by LC-MS/MS) (150 –299.9 ng/mL)	Near Cutoff Positive(Between the cutoffand 50% above thecutoff concentrationas determined by LC-MS/MS) (300 – 450ng/mL)	High Positives(Greater than 50%above cutoffconcentration (>450 ng/mL)
Positive	0	7*	17*	9	48
Negative	42	3	10	0	0

Discordant samples

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PreviousID	New ID	Immunoassay		LC-MS/MS (ng/mL)
		Qualitativemode	Semiquantitativemode	Hydrocodone	Hydromorphone	Hydromorphone3β-glucuronide
4	46	Positive	Positive	126	<LLOQ**	179
11	47	Positive	Positive	132	<LLOQ	160
15	48	Positive	Positive	126	<LLOQ	314
17	49	Positive	Positive	82.7	<LLOQ	128
18	50	Positive	Positive	138	<LLOQ	677
19	51	Positive	Positive	131	<LLOQ	126
29	52	Positive	Positive	73.3	<LLOQ	262
28	63	Positive	Positive	162	<LLOQ	218
31	64	Positive	Positive	249	<LLOQ	356
32	65	Positive	Positive	253	<LLOQ	265
33	66	Positive	Positive	160	<LLOQ	739
34	67	Positive	Positive	270	<LLOQ	88.6
35	68	Positive	Positive	217	<LLOQ	158
40	69	Positive	Positive	236	<LLOQ	90.4
49	70	Positive	Positive	282	<LLOQ	242
51	71	Positive	Positive	295	<LLOQ	433
59	72	Positive	Positive	167	<LLOQ	149
3	73	Positive	Positive	193	<LLOQ	756
10	74	Positive	Positive	214	<LLOQ	434
12	75	Positive	Positive	207	<LLOQ	706
13	76	Positive	Positive	298	<LLOQ	190
14	77	Positive	Positive	198	<LLOQ	262
21	78	Positive	Positive	157	<LLOQ	124
22	79	Positive	Positive	287	<LLOQ	79.9

* Discordant Result Table for Discrepant Sample near cutoff

Samples are discordant because of cross-reactivity of immunoassay to hydromorphone and hydromorphone-38glucuronide.

** Lower Limit of Quantitation (LLOQ) is 40 ng/mL.

e) Specificity The cross-reactivity of DRI Hydrocodone and its metabolites, opiate compounds and structurally related and unrelated compounds are evaluated by adding known amounts of each analyte to drug-free negative urine. In both qualitative and semi-quantitative modes, the assay cross-reacted with Hydrocodone at 300 ng/mL (100% cross-reactivity), Hydromorphone at 325 ng/mL (92% cross-reactivity), Norhydrocodone at 13,000 ng/mL (2% cross-reactivity), Dihydrocodeine at 12,500 ng/mL (2% cross-reactivity). In semi-quantitative mode the assay cross-reacted with Hydromorphone-3ß-glucuronide at 185 ng/mL (162% cross-reactivity).
In both qualitative and semi-quantitative modes, the assay exhibited minimal cross-reactivity with opiates and structurally related compounds at the tested concentrations.

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In both qualitative and semi-quantitative modes, the assay produced a negative result when structurally unrelated compounds were spiked into 225 ng/mL of hydrocodone (Low control level), at the tested concentrations. In both qualitative and semi-quantitative modes, the assay produced a positive result when structurally unrelated compounds were spiked into 375 ng/mL of hydrocodone (High control level) at the tested concentrations. These results indicated there was no cross-reactivity of the assay to the structurally unrelated compounds.

Hydrocodone and itsMetabolites	Tested Concentration(ng/mL)	Response Equivalentto Cutoff(Positive/Negative)	% Cross-Reactivity
Hydrocodone	250	Negative
	275	Negative
	300	Positive	100
	325	Positive
	350	Positive
Hydromorphone	200	Negative
	250	Negative
	300	Negative	92
	325	Positive
	350	Positive
	400	Positive
Hydromorphone-3β-glucuronide	175	Positive
	185	Positive
	200	Positive
	250	Positive	162
	300	Positive
	350	Positive
	400	Positive
Norhydrocodone	10,000	Negative
	11,000	Negative
	12,000	Negative
	13,000	Positive	2
	14,000	Positive
	15,000	Positive
6-Hydroxycodol(Dihydrocodeine)	11,000	Negative
	12,000	Negative
	12,500	Positive	2
	13,000	Positive
Opiates and Structurally RelatedCompounds	TestedConcentration(ng/mL)	ResponseEquivalent toCutoff(Positive/Negative)	% CrossReactivity
6-Acetyl morphine	100,000	Negative	< 0.3
Buprenorphine	100,000	Negative	< 0.3
Buprenorphone 3β-D glucuronide	100,000	Negative	< 0.3
Codeine	150,000	Negative	< 0.2
Dextromethorphan	250,000	Negative	< 0.1
EDDP	150,000	Negative	< 0.2
Fentanyl	100,000	Negative	< 0.3
Heroin	100,000	Negative	< 0.3
Levorphanol	18,000	Negative	1.4
	20,000	Negative
	22,000	Positive
Methadone	100,000	Negative	< 0.3
Meperidine	100,000	Negative	< 0.3
Morphine	150,000	Negative	< 0.2
Morphine-3β-glucuronide	70,000	Negative	< 0.4
Morphine-6β-glucuronide	75,000	Negative	< 0.4
Nalbuphine	150,000	Negative	< 0.2
Naloxone	15,000	Negative	1.8
	17,000	Positive
Naltrexone	50,000	Negative	0.4
	75,000	Positive
	100,000	Positive
Norbuprenorphine	100,000	Negative	< 0.3
Norcodeine	150,000	Negative	< 0.2
Normorphine	150,000	Negative	< 0.2
Noroxycodone	50,000	Negative	0.3
	100,000	Negative
	110,000	Positive
Oxycodone	8,000	Negative	2.1
	12,000	Negative
	14,000	Positive
Oxymorphone-β-D-glucuronide	8,000	Negative	2.1
	14,000	Positive
Oxymorphone	8,000	Negative	2.1
	12,0000	Negative
	14,0000	Positive
Opiates and Structurally RelatedCompounds	TestedConcentration(ng/mL)	ResponseEquivalent toCutoff(Positive/Negative)	% CrossReactivity
Tapentadol	100,000	Negative	< 0.3
Thebaine	100,000	Negative	< 0.3
Tramadol	100,000	Negative	< 0.3

Cross-reactivity of Hydrocodone and its metabolites

Opiates and structurally related compounds

Drug free urine was spiked with the indicated concentrations of compounds. Samples were tested in duplicates in both qualitative and semi-quantitative modes.

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Results in qualitative mode are expressed where a negative number indicates rates below cutoff calibrator (negative result), and a positive number indicates rates above cutoff calibrator (positive result). The results are shown in the table below.

Cross-reactivity of Opiates and structurally related compounds

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Structurally unrelated compounds

Structurally unrelated compounds were spiked into drug free urine that has been spiked to 225 ng/mL or 375 ng/mL of hydrocodone.

Samples were tested in 5 replicates in both qualitative and semi-quantitative modes. Results in qualitative mode are expressed as Δ Rate, where a negative number indicates rates below cutoff calibrator (negative result), and a positive number indicates rates above cutoff calibrator (positive result).

The results are shown in the table below.

Cross-Reactivity of structurally unrelated compounds
Compounds	TestedConcentration.(ng/mL)	Spiked Hydrocodone Level
		Low Control -25% ofCutoff (225 ng/mL)Positive/Negative	Low Control 25% ofCutoff (375 ng/mL)Positive/Negative
Negative Urine	0	Negative	Positive
Acetaminophen	500,000	Negative	Positive
Acetylsalicylic acid	500,000	Negative	Positive
Amitriptyline	100,000	Negative	Positive
Amoxicillin	100,000	Negative	Positive
Amphetamine	1,000,000	Negative	Positive
Benzoylecgonine	1,000,000	Negative	Positive
Caffeine	100,000	Negative	Positive
Carbamazepine	500,000	Negative	Positive
Chlorpromazine	100,000	Negative	Positive
Clomipramine	10,000	Negative	Positive
Cimetidine	500,000	Negative	Positive
Desipramine	100,000	Negative	Positive
Diphenhydramine	100,000	Negative	Positive
Doxepin	100,000	Negative	Positive
Ephedrine	1,000,000	Negative	Positive
Fluoxetine	100,000	Negative	Positive
Fluphenazine	100,000	Negative	Positive
Ibuprofen	500,000	Negative	Positive
Imipramine	100,000	Negative	Positive
Maprotiline	100,000	Negative	Positive
Nortriptyline	100,000	Negative	Positive

Cross-reactivity of structurally unrelated compounds

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Compounds	TestedConcentration.(ng/mL)	Spiked Hydrocodone Level
		Low Control -25% ofCutoff (225 ng/mL)Positive/Negative	Low Control 25% ofCutoff (375 ng/mL)Positive/Negative
Oxazepam	250,000	Negative	Positive
Phencyclidine (PCP)	100,000	Negative	Positive
Phenobarbital	100,000	Negative	Positive
Ranitidine	500,000	Negative	Positive
Secobarbital	100,000	Negative	Positive
Thioridazine	100,000	Negative	Positive

f) Interference The potential interference of pH and endogenous physiologic substances on recovery of Hydrocodone using DRI Hydrocodone Assay is assessed by spiking known compounds of potentially interfering substances into the Low Control (225 ng/mL cutoff) and High Control (375 ng/mL cutoff). In the presence of the compounds listed below, the controls are detected accurately, indicating that these compounds did not show interference in the assay.

Interference substances

Compounds	TestedConcentration.(mg/dL)	Spiked Hydrocodone Level
		Low Control -25% ofcutoff (225 ng/mL)	High Control 25% ofcutoff (375 ng/mL)
		Positive/ Negative	Positive/Negative
Negative Urine	0	Negative	Positive
Acetaminophen	10	Negative	Positive
Acetone	500	Negative	Positive
Acetyl Salicylic Acid	10	Negative	Positive
Ascorbic Acid	150	Negative	Positive
Caffeine	10	Negative	Positive
Creatinine	400	Negative	Positive
Ethanol	10	Negative	Positive
Galactose	5	Negative	Positive
Glucose	1000	Negative	Positive
Hemoglobin	150	Negative	Positive
Human Serum Albumin	200	Negative	Positive
Ibuprophen	10	Negative	Positive
Oxalic acid	50	Negative	Positive
Riboflavin	3	Negative	Positive
Sodium Chloride	1000	Negative	Positive
Urea	1000	Negative	Positive

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pH samples

Compound	Tested Concentration	Spiked Hydrocodone Level
		Low Control -25% of cutoff (225 ng/mL) Positive/Negative	High Control 25% of cutoff (375 ng/mL) Positive/Negative
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

h) Specific Gravity Drug free urine samples with specific gravity ranging in value within 1.003 to 1.031 are split and spiked to a final concentration of either 225ng/mL or 375ng/mL (the Low Control and High Concentrations, respectively). These samples are then evaluated in both qualitative and semi-quantitative modes. No interference is observed.

Specific gravity samples

Specificgravity	Spiked Hydrocodone Level
	Low Control -25% of cutoff (225ng/mL)Positive/Negative	High Control 25% of cutoff (375ng/mL)Positive/Negative
1.003	Negative	Positive
1.005	Negative	Positive
1.006	Negative	Positive
1.010	Negative	Positive
1.011	Negative	Positive
1.014	Negative	Positive
1.019	Negative	Positive
1.024	Negative	Positive
1.027	Negative	Positive
1.031	Negative	Positive

i) Traceability

The primary controls and calibrators are traceable to the 1 mg/mL Hydrocodone stock solution purchased from a commercial source which is established at 99.9% purity. The concentration of the primary control and calibrator stocks is confirmed by LC-MS/MS from three independent laboratories.

g) pH

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j) Stability

Open Vial Stability:

Open Vial stability studies for two lots stored at 2-8°C supports the claim of 60 days for qualitative and semi-quantitative modes. This data was presented in the original 510(k) submission K150502

Reagent On-Board Stability:

Reagent On-Board stability studies for one lot stored on-board clinical analyzer supports the claim of 60 days for qualitative and semi-quantitative modes.

Real Time Stability for Reagent

Real Time stability testing results show that the Low Control is detected as negative and the High Control is detected as positive for each time point for a period of 2 year at 2-8℃. The recoveries of the Low Control and High Control are within 80-120%. Our current shelf-life claim is 18 months.

k) Detection Limit Limit of detection is not provided in a 510(k) of this type of assay.

l) Clinical Studies

a. Clinical Sensitivity Clinical sensitivity is not provided in a 510(k) of this type of assay.
b. Clinical Specificity Clinical specificity is not provided in a 510(k) of this type of assay.
m) Clinical cutoff There are currently no SAMHSA recommendations for a clinical cutoff for hydrocodone as of today.

H. Conclusion

The information supports a determination of substantial equivalence between DRI Hydrocodone on Indiko Plus and DRI Hydrocodone Assay, Calibrators and Control Set on AU 680 (K150502).

Regulation Number and Section

§ 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).