(147 days)
The ARK Hydrocodone Assay is an immunoassay intended for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Hydrocodone Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
The ARK Hydrocodone Assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay is based on competition between hydrocodone in the specimen and hydrocodone labeled with recombinant glucose-o-phosphate dehydrogenase (rG6PDH) for antibody binding sites. As the latter binds antibody, enzyme activity decreases. In the presence of hydrocodone from the specimen, enzyme activity increases and is directly related to the hydrocodone concentration. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay.
The ARK Hydrocodone Assay consists of reagents R1 anti-hydrocodone monoclonal rabbit antibodies with substrate and R2 hydrocodone derivative labeled with bacterial recombinant G6PDH enzyme.
The provided document is a 510(k) summary for the ARK Hydrocodone Assay, an immunoassay for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine.
Here's an analysis of the acceptance criteria and the study that proves the device meets them, based on the provided text:
1. Table of Acceptance Criteria & Reported Device Performance:
The document doesn't explicitly define formal "acceptance criteria" in a separate table with numerical thresholds for accuracy, precision, etc. Instead, it presents various performance characteristics that collectively demonstrate the device's suitability for its stated intended use and substantial equivalence to a predicate device. The implied acceptance criteria are that the device performs reliably and similarly to the predicate in key analytical measures.
Below is a table summarizing the reported device performance, which implicitly functions as the evidence meeting the "acceptance criteria" for analytical validation.
| Performance Characteristic | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Precision (Qualitative) | Consistent classification (Positive/Negative) at specified concentrations relative to the 300 ng/mL cutoff, especially near the cutoff. | 0 ng/mL: 160 Negative (N=160)75 ng/mL: 160 Negative (N=160)150 ng/mL: 160 Negative (N=160)225 ng/mL (-25% Cutoff): 160 Negative (N=160)300 ng/mL (Cutoff): 50 Negative / 110 Positive (N=160)375 ng/mL (+25% Cutoff): 160 Positive (N=160)450 ng/mL (+50% Cutoff): 160 Positive (N=160)525 ng/mL (+75% Cutoff): 160 Positive (N=160)600 ng/mL (+100% Cutoff): 160 Positive (N=160) |
| Precision (Semi-quantitative) | Consistent mean recovery and classification at specified concentrations relative to the 300 ng/mL cutoff. | 0 ng/mL: Mean 0, 160 Negative75 ng/mL: Mean 78, 160 Negative150 ng/mL: Mean 142, 160 Negative225 ng/mL: Mean 229, 160 Negative300 ng/mL (Cutoff): Mean 314, 24 Neg / 136 Pos375 ng/mL: Mean 388, 160 Positive450 ng/mL: Mean 459, 160 Positive525 ng/mL: Mean 539, 160 Positive600 ng/mL: Mean 620, 160 Positive |
| Analytical Recovery | Percent recovery close to 100% across the assay range. | Ranges from 86% to 103% (e.g., 80 ng/mL expected yielded 99% recovery, 720 ng/mL yielded 86% recovery, 800 ng/mL yielded 92% recovery). |
| Analytical Specificity (Cross-reactivity to Metabolites) | Cross-reactivity for hydrocodone should be high, while for metabolites it should be understood and ideally lower if they are not the primary target the assay is trying to quantify. | Hydrocodone: 103%Hydromorphone: 100%Hydromorphone-3β-Glucuronide: 0.7%Norhydrocodone: 13.2%Dihydrocodeine: <0.3% (at >100,000 ng/mL tested) |
| Analytical Specificity (Cross-reactivity to Structurally Related/Unrelated Compounds) | No significant cross-reactivity with other common opiate compounds or structurally unrelated substances should lead to false positives at specified concentrations. | All 30 tested structurally related or unrelated opiate compounds (e.g., Morphine, Codeine, Fentanyl, Oxycodone, Methadone, Buprenorphine, Heroin) showed NEGATIVE results at concentrations up to 100,000 ng/mL, with cross-reactivity generally <0.3% (or <0.6% for some at 50,000 ng/mL). |
| Interference (Endogenous Substances) | No interference (maintaining correct POS/NEG classification) from high concentrations of common endogenous substances. | All 17 tested endogenous substances (e.g., Acetaminophen, Creatinine, Glucose, Hemoglobin, Urea) showed no interference, maintaining NEG for 225 ng/mL hydrocodone and POS for 375 ng/mL hydrocodone. |
| Interference (Boric Acid) | No interference from boric acid. | Boric Acid (1% w/v) showed no interference, maintaining NEG for 225 ng/mL hydrocodone and NEG for 375 ng/mL hydrocodone (this second result seems like a typo in the table if it should be POS, but is recorded as NEG). Correction needed for interpretation of the Boric Acid result for 375 ng/mL hydrocodone. Given the other interference studies, it is likely it should have been POS, or implies an interfering effect with boric acid at the higher concentration. This would require clarification. |
| Specific Gravity Interference | No interference across a physiological range of urine specific gravity. | No interference observed from urine samples with specific gravity values ranging from 1.000 to 1.030, maintaining NEG for 225 ng/mL hydrocodone and POS for 375 ng/mL hydrocodone. |
| pH Interference | No interference across a physiological range of urine pH. | No interference observed from urine samples with pH values from 3.0 to 11.0, maintaining NEG for 225 ng/mL hydrocodone and POS for 375 ng/mL hydrocodone. |
| Method Comparison (Overall Concordance with LC-MS/MS) | High overall concordance with the confirmatory method (LC-MS/MS). | Overall concordance: 92.5% |
| Method Comparison (Qualitative Performance) | High sensitivity and specificity, especially around the cutoff. Discordant results should be explainable. | True Negatives: 134 samples (<150 ng/mL by LC-MS/MS) correctly identified as Negative. True Positives: 9 samples (300-450 ng/mL by LC-MS/MS) and 66 samples (>450 ng/mL by LC-MS/MS) correctly identified as Positive. Discordant Results: 8 samples (<150 ng/mL) and 8 samples (150-299 ng/mL) were assay Positive but LC-MS/MS Negative. 1 sample (300-450 ng/mL) was assay Negative but LC-MS/MS Positive. Discordances are explained as cross-reactivity with hydromorphone. |
2. Sample Sizes Used for the Test Set and Data Provenance:
- Precision Studies: N=160 for both qualitative and semi-quantitative precision (quadruplicate assays twice a day for 20 days for 8 concentration levels).
- Analytical Recovery: 5 replicates for each of the 11 concentration points across the assay range.
- Analytical Specificity (Cross-reactivity): Concentrations tested for various compounds (e.g., 100,000 ng/mL, 50,000 ng/mL). The exact number of individual tests/replicates is not specified but implied testing at least one sample per compound.
- Interference (Endogenous Substances, Boric Acid, Specific Gravity, pH): Not explicitly stated, but for Specific Gravity and pH, "N=3" is mentioned for each condition, suggesting triplicate testing.
- Method Comparison (Clinical Specimens): 226 unaltered clinical urine specimens.
- Data Provenance: The document states that the clinical urine specimens for method comparison were "unaltered clinical urine specimens that are not individually identifiable." It does not specify the country of origin or if they were retrospectively or prospectively collected. The analytical studies (precision, recovery, specificity, interference) appear to be laboratory-based studies using spiked drug-free human urine.
3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications:
This information is not provided in the document. The ground truth for the clinical specimens in the method comparison study was established by LC-MS/MS (Liquid Chromatography/tandem Mass Spectrometry), which is stated as the "preferred confirmatory method." LC-MS/MS is an analytical chemistry technique, not a human expert interpretation.
4. Adjudication Method for the Test Set:
This information is not applicable/provided. The ground truth was established by LC-MS/MS, an objective analytical method. Therefore, there was no need for human expert adjudication.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done:
No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. This device is an immunoassay (a laboratory diagnostic test), not an imaging AI or a device that assists human readers in interpreting complex data. The performance study compares the device's analytical results to a gold standard analytical method (LC-MS/MS), not a human's performance.
6. If a Standalone (Algorithm Only Without Human-In-The-Loop Performance) was done:
The device itself is a "standalone" test in the sense that it provides a direct analytical result (qualitative or semi-quantitative) without human interpretation of raw data for diagnosis. The study presented (precision, analytical recovery, specificity, method comparison) evaluates the algorithm's (assay's) performance independent of direct human-in-the-loop diagnostic assistance. While a human operates the instrument and interprets the final result, the assay's core function is automated chemical analysis.
7. The Type of Ground Truth Used:
The primary ground truth used for the method comparison study was LC-MS/MS (Liquid Chromatography/tandem Mass Spectrometry) results. This is considered a highly specific and sensitive "confirmatory method" in analytical chemistry, often serving as a gold standard for drug quantification. For the other analytical studies (precision, recovery, specificity, interference), the ground truth was the known concentrations of spiked analytes into drug-free urine.
8. The Sample Size for the Training Set:
This information is not applicable/provided. This is an immunoassay, not a machine learning/AI model that requires a "training set" in the conventional sense. The "training" for such a device occurs during its development and optimization of the reagents and assay parameters by the manufacturer, rather than through a dataset in a machine learning paradigm.
9. How the Ground Truth for the Training Set Was Established:
As mentioned above, this concept doesn't directly apply here. The "ground truth" during the development and optimization of the assay would have been established through controlled laboratory experiments using known concentrations of hydrocodone and related substances, verified perhaps by independent analytical methods (e.g., HPLC, LC-MS/MS) and chemical purity assessments. The goal is to optimize the assay's sensitivity, specificity, and precision to meet performance targets.
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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 acronym along with the full name of the agency on the right. The FDA part of the logo is in blue, with the acronym in a square and the full name written out to the right of it.
ARK Diagnostics, Inc. Thomas Houts, Ph.D. Sr. Director, Quality, Regulatory and Planning 48089 Fremont Boulevard Fremont, California 94538
Re: K231752
Trade/Device Name: ARK Hydrocodone Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: Class II Product Code: DJG Dated: October 6, 2023 Received: October 6, 2023
Dear Thomas Houts:
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 (the 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 available 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
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Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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, Joseph A. Digitally signed by Kotarek -S Date: 2023.11.09 Joseph Kotarek, Ph.D. Toxicology Branch Chief Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K231752
Device Name ARK Hydrocodone Assay
Indications for Use (Describe)
The ARK Hydrocodone Assay is an immunoassay intended for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL. The semi-quantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Hydrocodone Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
Type of Use (Select one or both, as applicable)
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) |
|---|
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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Section 05: 510(k) SUMMARY
This 510(k) Summary of Safety and Effectiveness information is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.
The assigned 510(k) number is K231752.
| 807.92 (a)(1): Name: | ARK Diagnostics, Inc. |
|---|---|
| Address: | 48089 Fremont BlvdFremont, CA 94538 USA |
| Owner Operator Number: | 10027663 |
| Establishment Registration: | 3005755244 |
| Phone: | (510) 270-6270 |
| FAX: | (510) 270-6298 |
| Contact: | Thomas Houts, Ph.D., FAACCARK Diagnostics, Inc.Director, Quality, Regulatory and PlanningTelephone: (510) 270-6296E-mail: tom@ark-tdm.com |
Date Prepared: June 5, 2023
807.92 (a)(2): Device Name – Trade Name, Common Name, and Classification
| Trade Name: | ARK Hydrocodone Assay |
|---|---|
| Common Name: | Homogeneous Enzyme Immunoassay, Opiate Test System |
| Classification: |
| Product Code | Classification | Regulation Section | Panel |
|---|---|---|---|
| DJG | Class II | 21 CFR 862.3650Opiate Test System | Toxicology(91) |
807.92 (a)(3): Identification of the Legally Marketed Predicate Device
DRI Hydrocodone Assay (K150502)
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807.92 (a)(4): Device Description
The ARK Hydrocodone Assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay. The assay is based on competition between hydrocodone in the specimen and hydrocodone labeled with recombinant glucose-o-phosphate dehydrogenase (rG6PDH) for antibody binding sites. As the latter binds antibody, enzyme activity decreases. In the presence of hydrocodone from the specimen, enzyme activity increases and is directly related to the hydrocodone concentration. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay.
The ARK Hydrocodone Assay consists of reagents R1 anti-hydrocodone monoclonal rabbit antibodies with substrate and R2 hydrocodone derivative labeled with bacterial recombinant G6PDH enzyme.
807.92 (a)(5): Intended Use / Indications for Use
ARK Hydrocodone Assay
The ARK Hydrocodone Assay is an immunoassay intended for the qualitative detection and/or semi-quantitative estimation of hydrocodone and its metabolites in human urine at a cutoff of 300 ng/mL.
The semi-quantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.
The ARK Hydrocodone Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.
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807.92 (a)(6): Technological Similarities and Differences to the Predicate
SUBSTANTIAL EQUIVALENCE COMPARATIVE TABLE
Comparison between the DRI Hydrocodone Assay and the ARK Hydrocodone Assay
| Characteristic | Predicate Device | Candidate Device |
|---|---|---|
| Similarities | ||
| Test System | Homogenous enzyme immunoassay(HEIA) | Same |
| Intended Use | The DRI® Hydrocodone Assay isintended for the qualitative and semi-quantitative detection and estimationof Hydrocodone and its metabolites inhuman urine at a cutoff of 300 ng/mL.The semi-quantitative mode is forpurposes of enabling laboratories todetermine an appropriate dilution ofspecimen for confirmation by aconfirmatory method such as LC-MS/MS or GCMS and permittinglaboratories toestablish quality control measures.This assay provides a preliminaryanalytical test result. A more specificalternative chemical method must beused in order toconfirm an analytical result. Gaschromatography/mass spectrometry(GC/MS) and LiquidChromatography/tandem massspectrometry (LC-MS/MS) are thepreferred confirmatory methods.Clinical consideration andprofessional judgmentshould be applied to any drug ofabuse test result, particularly whenpreliminary positive results are used. | Same |
| Sample Matrix | Human urine | Same |
| User Environment | Clinical laboratory; Prescription useonly | Same |
| MassSpectrometryConfirmation | Required to confirm preliminarypositive analytical results | Same |
| Platform Required | Automated clinical chemistryanalyzer | Same |
| Reagents Form | Liquid - Ready-to-use | Same |
| Reagent Materials | Two (2) reagent system:Antibody/substrate reagent (mousemonoclonal antibodies tohydrocodone) and enzyme labeledconjugate (hydrocodone derivativelabeled with enzyme)Sodium azide preservative | Two (2) reagent system:Antibody/substrate reagent (rabbitmonoclonal antibodies tohydrocodone) and enzyme labeledconjugate (hydrocodone derivativelabeled with enzyme)Sodium azide preservative |
| Quality Controls | Two levels - 225 and 375 ng/mL | Same |
| Storage | 2-8°C until expiration date | Same |
| Measured Analyte | Hydrocodone | Same |
| Detection | Absorbance change measuredspectrophotometrically at 340 nm | Same |
| Cutoff Level | 300 ng/mL | Same |
| Control Levels | 225 and 375 ng/mL | Same |
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| Characteristic | Predicate DeviceDRI Hydrocodone Assay (K150502) | Candidate DeviceARK Hydrocodone Assay |
|---|---|---|
| Differences | ||
| Antibody | Mouse monoclonal | Rabbit monoclonal |
| Calibrator Levels | 0, 100, 300, 500, 1000 ng/mL | 0, 50, 100, 300, 800 ng/mL |
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807.92 (b)(1) and 807.92 (b)(2): Brief Description of Nonclinical and Clinical Data
The following performance characteristics were obtained on the Beckman Coulter AU680® automated clinical chemistry analyzer.
Precision
Precision studies were performed using CLSI EP05-A3 as a guideline. Drug-free, negative human urine was supplemented with hydrocodone (0 to 600 ng/mL). Each level was assayed in quadruplicate twice a day for 20 days (N=160) and evaluated qualitatively and semiquantitatively. Results are summarized in the tables below.
| Hydrocodone(ng/mL) | Relative %Cutoff | # of Results | Precision Results |
|---|---|---|---|
| 0 | -100 | 160 | 160 Neg |
| 75 | -75 | 160 | 160 Neg |
| 150 | -50 | 160 | 160 Neg |
| 225 | -25 | 160 | 160 Negative |
| 300 | Cutoff | 160 | 50 Neg /110 Pos |
| 375 | +25 | 160 | 160 Pos |
| 450 | +50 | 160 | 160 Pos |
| 525 | +75 | 160 | 160 Pos |
| 600 | +100 | 160 | 160 Pos |
Qualitative Precision
Semi-quantitative Precision
| Hydrocodone(ng/mL) | Relative%Cutoff | # ofResults | Mean(ng/mL) | PrecisionResults |
|---|---|---|---|---|
| 0 | -100 | 160 | 0 | 160 Neg |
| 75 | -75 | 160 | 78 | 160 Neg |
| 150 | -50 | 160 | 142 | 160 Neg |
| 225 | -25 | 160 | 229 | 160 Neg |
| 300 | Cutoff | 160 | 314 | 24 Neg /136 Pos |
| 375 | +25 | 160 | 388 | 160 Pos |
| 450 | +50 | 160 | 459 | 160 Pos |
| 525 | +75 | 160 | 539 | 160 Pos |
| 600 | +100 | 160 | 620 | 160 Pos |
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Analytical Recovery
Drug-free, negative human urine was spiked with hydrocodone across the assay range of the semi-quantitative calibration curve. Each sample was run in replicates of 5 in semi-quantitative mode and the average was used to determine percent recovery compared to the expected value.
| Expected Value(ng/mL) | Observed Value(ng/mL) | Recovery (%) |
|---|---|---|
| 0 | 0 | N/A |
| 80 | 80 | 99 |
| 160 | 151 | 94 |
| 240 | 247 | 103 |
| 320 | 322 | 101 |
| 400 | 386 | 96 |
| 480 | 472 | 98 |
| 560 | 537 | 96 |
| 640 | 606 | 95 |
| 720 | 621 | 86 |
| 800 | 738 | 92 |
Analytical Specificity
All compounds tested were added to drug-free, negative human urine and tested with the ARK Hydrocodone Assay in both qualitative and semi-quantitative modes.
The cross-reactivity of hydrocodone and its metabolites was evaluated by spiking these compounds into drug-free, negative human urine and evaluated by dose-response to determine the approximate equivalence to the 300 ng/mL hydrocodone cutoff. These concentrations were used to determine the percent cross-reactivity according to the formula:
% Cross-reactivity = (Cutoff concentration / Concentration approximately equivalent to the 300 ng/mL cutoff) X 100
For compounds that did not produce a positive result, the highest concentration tested was used to calculate percent cross-reactivity.
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| Compound | ConcentrationApproximatelyEquivalent to theCutoff(ng/mL) | Cross-reactivity (%) |
|---|---|---|
| Hydrocodone | 292 | 103 |
| Hydromorphone | 299 | 100 |
| Hydromorphone-3β-Glucuronide | 45,439 | 0.7 |
| Norhydrocodone | 2,277 | 13.2 |
| Dihydrocodeine | >100,000 | <0.3 |
Cross-reactivity of hydrocodone and its metabolites
Cross-reactivity of structurally related or unrelated opiate compounds
| Compound | ConcentrationTested(ng/mL) | POS/NEG | Cross-reactivity (%) |
|---|---|---|---|
| 6-Acetyl morphine | 100,000 | NEG | <0.3 |
| Buprenorphine | 100,000 | NEG | <0.3 |
| Buprenorphine-3β-D-glucuronide | 50,000 | NEG | <0.6 |
| Codeine | 100,000 | NEG | <0.3 |
| Codeine-6β-D-glucuronide | 100,000 | NEG | <0.3 |
| Dextromethorphan | 250,000 | NEG | <0.1 |
| EDDP | 100,000 | NEG | <0.3 |
| EMDP | 100,000 | NEG | <0.3 |
| Ethyl morphine | 100,000 | NEG | <0.3 |
| Fentanyl | 100,000 | NEG | <0.3 |
| Heroin | 100,000 | NEG | <0.3 |
| Levorphanol | 100,000 | NEG | <0.3 |
| Meperidine | 100,000 | NEG | <0.3 |
| Methadone | 100,000 | NEG | <0.3 |
| Morphine | 100,000 | NEG | <0.3 |
| Morphine-3β-D-glucuronide | 100,000 | NEG | <0.3 |
| Morphine-6β-D-glucuronide | 100,000 | NEG | <0.3 |
| Nalbuphine | 100,000 | NEG | <0.3 |
| Naloxegol | 100,000 | NEG | <0.3 |
| Naloxone | 100,000 | NEG | <0.3 |
| Naltrexone | 100,000 | NEG | <0.3 |
| Norbuprenorphine | 100,000 | NEG | <0.3 |
| Norcodeine | 100,000 | NEG | <0.3 |
| Normorphine | 100,000 | NEG | <0.3 |
| Noroxycodone | 100,000 | NEG | <0.3 |
| Nortilidine | 100,000 | NEG | <0.3 |
| Oxycodone | 100,000 | NEG | <0.3 |
| Oxymorphone | 100,000 | NEG | <0.3 |
| Oxymorphone-3β-D-glucuronide | 50,000 | NEG | <0.6 |
| Pentazocine | 100,000 | NEG | <0.3 |
| Tapentadol | 100,000 | NEG | <0.3 |
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| Thebaine | 100,000 | NEG | <0.3 |
|---|---|---|---|
| Tilidine | 100,000 | NEG | <0.3 |
| Tramadol | 100,000 | NEG | <0.3 |
| Compound | ConcentrationTested(ng/mL) | POS/NEG |
|---|---|---|
| (+)-MDA | 100,000 | NEG |
| 11-hydroxy-delta-9-THC | 100,000 | NEG |
| 11-nor-9 carboxy THC | 50,000 | NEG |
| 1R,2S(-)-Ephedrine | 100,000 | NEG |
| 1S,2R(+)-Ephedrine | 100,000 | NEG |
| 4-Bromo-2,5,Dimethoxyphenethylamine | 100,000 | NEG |
| 7-Aminoclonazepam | 100,000 | NEG |
| Acetaminophen | 500,000 | NEG |
| Acetylsalicylic acid | 500,000 | NEG |
| Alprazolam | 100,000 | NEG |
| Amitriptyline | 100,000 | NEG |
| Amobarbital | 100,000 | NEG |
| Amoxicillin | 100,000 | NEG |
| Amphetamine | 100,000 | NEG |
| Atorvastatin | 100,000 | NEG |
| Benzoylecgonine | 1,000,000 | NEG |
| Benzylpiperazine | 100,000 | NEG |
| Bupropion | 100,000 | NEG |
| Butabarbital | 100,000 | NEG |
| Caffeine | 100,000 | NEG |
| Canagliflozin | 50,000 | NEG |
| Cannabidiol | 100,000 | NEG |
| Cannabinol | 100,000 | NEG |
| Carbamazepine | 500,000 | NEG |
| Carisoprodol | 100,000 | NEG |
| Chlordiazepoxide | 100,000 | NEG |
| Chlorpromazine | 100,000 | NEG |
| Cimetidine | 500,000 | NEG |
| Clobazam | 100,000 | NEG |
| Clomipramine | 100,000 | NEG |
| Clopidogrel | 100,000 | NEG |
| Cocaine | 100,000 | NEG |
| Cotinine | 100,000 | NEG |
| Cyclobenzaprine | 100,000 | NEG |
| Desipramine | 100,000 | NEG |
| Diazepam | 100,000 | NEG |
| Diphenhydramine | 100,000 | NEG |
| Doxepin | 100,000 | NEG |
| Ecgonine | 100,000 | NEG |
| Ephedrine | 1,000,000 | NEG |
| Fluoxetine | 100,000 | NEG |
| Fluphenazine | 100,000 | NEG |
| Ibuprofen | 500,000 | NEG |
| Imipramine | 100,000 | NEG |
| Ketamine | 100,000 | NEG |
| Lamotrigine | 100,000 | NEG |
| Lidocaine | 100,000 | NEG |
| LSD | 100,000 | NEG |
| Maprotiline | 100,000 | NEG |
| MDMA | 50,000 | NEG |
| Meprobamate | 100,000 | NEG |
| Metformin | 100,000 | NEG |
| Methylphenidate | 250,000 | NEG |
| Metronidazole | 100,000 | NEG |
| Naproxen | 100,000 | NEG |
| Norpseudoephedrine | 50,000 | NEG |
| Nortriptyline | 100,000 | NEG |
| Omeprazole | 100,000 | NEG |
| Ondansetron | 100,000 | NEG |
| Oxazepam | 250,000 | NEG |
| Phencyclidine | 100,000 | NEG |
| Phenobarbital | 100,000 | NEG |
| Phentermine | 100,000 | NEG |
| Phenylephrine | 100,000 | NEG |
| Phenylpropanolamine | 100,000 | NEG |
| Phenytoin | 100,000 | NEG |
| PMA | 100,000 | NEG |
| Propranolol | 100,000 | NEG |
| Protriptyline | 100,000 | NEG |
| R,R(-)-Pseudoephedrine | 100,000 | NEG |
| Ranitidine | 500,000 | NEG |
| Ritalinic Acid | 100,000 | NEG |
| S(+)-Methamphetamine | 100,000 | NEG |
| S,S(+)-Pseudoephedrine | 100,000 | NEG |
| Salicylic Acid | 100,000 | NEG |
| Secobarbital | 100,000 | NEG |
| Sertraline | 100,000 | NEG |
| Temazepam | 100,000 | NEG |
| Theophylline | 50,000 | NEG |
| Thioridazine | 100,000 | NEG |
| Trazodone | 100,000 | NEG |
| Triazolam | 250,000 | NEG |
| Trimipramine | 100,000 | NEG |
| Venlafaxine | 100,000 | NEG |
| Zolpidem | 100,000 | NEG |
Structurally unrelated compounds
{11}------------------------------------------------
{12}------------------------------------------------
Interference
Endogenous Substances
High concentrations of the following endogenous substances were added into hydrocodonespiked urine (± 25% of the cutoff concentration). No interference was observed when tested with the ARK Hydrocodone Assay.
| Compound | ConcentrationTested(mg/dL) | 225 ng/mL(-25% Cutoff) | 375 ng/mL(+25% Cutoff) |
|---|---|---|---|
| Acetaminophen | 10 | NEG | POS |
| Acetone | 500 | NEG | POS |
| Acetyl Salicylic Acid | 10 | NEG | POS |
| Ascorbic acid | 150 | NEG | POS |
| Caffeine | 10 | NEG | POS |
| Creatinine | 400 | NEG | POS |
| Ethanol | 10 | NEG | POS |
| Galactose | 5 | NEG | POS |
| Glucose | 1000 | NEG | POS |
| Hemoglobin | 150 | NEG | POS |
| Human Albumin | 200 | NEG | POS |
| Human y- Globulin | 500 | NEG | POS |
| Ibuprofen | 10 | NEG | POS |
| NaCl | 1000 | NEG | POS |
| Oxalic Acid | 50 | NEG | POS |
| Riboflavin | 3 | NEG | POS |
| Urea | 1000 | NEG | POS |
Interference – Boric Acid
One percent (1%) w/v of boric acid was added into hydrocodone-spiked urine (± 25% of the cutoff concentration). Results are provided in the table below.
| Compound | ConcentrationTested | 225 ng/mL(-25% Cutoff) | 375 ng/mL(+25% Cutoff) |
|---|---|---|---|
| Boric Acid | 1% w/v | NEG | NEG |
{13}------------------------------------------------
Specific Gravity
Urine samples with specific gravity values ranging from 1.000 to 1.030 were tested in the presence of the two levels of hydrocodone at ± 25% of the 300 ng/mL cutoff concentration. No interference was observed when tested with the ARK Hydrocodone Assay in both qualitative and semi-quantitative modes.
| Compound Tested | 225 ng/mLHydrocodoneN=3(POS/NEG) | 375 ng/mLHydrocodoneN=3(POS/NEG) |
|---|---|---|
| Specific Gravity 1.0000 | NEG | POS |
| Specific Gravity 1.0021 | NEG | POS |
| Specific Gravity 1.0043 | NEG | POS |
| Specific Gravity 1.0179 | NEG | POS |
| Specific Gravity 1.0187 | NEG | POS |
| Specific Gravity 1.0262 | NEG | POS |
| Specific Gravity 1.0303 | NEG | POS |
pH
Urine samples with pH values from 3.0 to 11.0 were tested in the presence of the two levels of hydrocodone at ± 25% of the 300 ng/mL cutoff concentration. No interference was observed when tested with the ARK Hydrocodone Assay in both qualitative and semi-quantitative modes.
| Compound Tested | 225 ng/mLHydrocodoneN=3(POS/NEG) | 375 ng/mLHydrocodoneN=3(POS/NEG) |
|---|---|---|
| Urine pH 3 | NEG | POS |
| Urine pH 4 | NEG | POS |
| Urine pH 5 | NEG | POS |
| Urine pH 6 | NEG | POS |
| Urine pH 7 | NEG | POS |
| Urine pH 8 | NEG | POS |
| Urine pH 9 | NEG | POS |
| Urine pH 10 | NEG | POS |
| Urine pH 11 | NEG | POS |
{14}------------------------------------------------
Method Comparison
Two hundred twenty-six (226) unaltered clinical urine specimens that are not individually identifiable were analyzed by ARK Hydrocodone Assay in both qualitative and semi-quantitative modes and the results were compared to LC-MS/MS. The overall concordance between LC-MS/MS and the ARK Hydrocodone Assay was 92.5%.
Qualitative method comparison with LC-MS/MS as reference method
| ARKHydrocodoneAssay Results | <50% of cutoffconcentration byLC-MS/MS(<150 ng/mL) | Near CutoffNegative(Between 50%below the cutoffandthe cutoffconcentration byLC-MS/MS)(150-299 ng/mL) | Near CutoffPositive(Between thecutoff and 50%above the cutoffconcentration byLC-MS/MS)(300-450 ng/mL) | High Positive(Greater than50% above thecutoffconcentration byLC-MS/MS)(>450 ng/mL) |
|---|---|---|---|---|
| Positive | 8* | 8* | 9 | 66 |
| Negative | 134 | 0 | 1* | 0 |
Semi-quantitative method comparison with LC-MS/MS as reference method
| ARKHydrocodoneAssay Results | <50% of cutoffconcentration byLC-MS/MS(<150 ng/mL) | Near CutoffNegative(Between 50%below the cutoffandthe cutoffconcentration byLC-MS/MS)(150-299 ng/mL) | Near CutoffPositive(Between thecutoff and 50%above the cutoffconcentration byLC-MS/MS)(300-450 ng/mL) | High Positive(Greater than50% above thecutoffconcentration byLC-MS/MS)(>450 ng/mL) |
|---|---|---|---|---|
| Positive | 8* | 8* | 9 | 66 |
| Negative | 134 | 0 | 1* | 0 |
*Discordant Results
{15}------------------------------------------------
| Sample # | ARKQualitative(POS/NEG) | LC-MS/MS (ng/mL) | ARKSemi-quantitative(ng/mL) | ||
|---|---|---|---|---|---|
| Hydrocodone | Hydromorphone | AdjustedTotal | |||
| 3 | POS | 287.8 | 210.0 | 497.8 | 476.0 |
| 15 | POS | 226.9 | 150.5 | 377.4 | 390.0 |
| 18 | POS | 156.0 | 174.7 | 330.8 | 335.2 |
| 23 | POS | 5.4 | 1317.7 | 1323.1 | 387.7 |
| 38 | NEG | 306.5 | 22.4 | 328.9 | 277.6 |
| 39 | POS | 162.0 | 52.4 | 214.5 | 316.1 |
| 48 | POS | 200.5 | 61.7 | 262.2 | 358.4 |
| 51 | POS | 174.4 | 29.0 | 203.4 | 357.4 |
| 66 | POS | 146.7 | 190.3 | 337.0 | 463.1 |
| 68 | POS | 181.2 | 150.3 | 331.5 | 382.2 |
| 70 | POS | Not Detected | 545.7 | 545.7 | 445.7 |
| 75 | POS | 5.9 | 10524.1 | 10530.0 | 2549.1 |
| 86 | POS | 255.8 | 30.7 | 286.5 | 471.4 |
| 90 | POS | 106.5 | 214.0 | 320.4 | 335.5 |
| 97 | POS | Not Detected | 1769.8 | 1769.8 | 501.3 |
| 99 | POS | Not Detected | 706.1 | 706.1 | 657.9 |
| 101 | POS | Not Detected | 5461.7 | 5461.7 | 2014.2 |
Seventeen (17) samples were considered discordant relative to the ARK Hydrocodone 300 ng/mL cutoff. Cross-reactivity to the major metabolite hydromorphone contributed to the positive results for samples with <300 ng/mL hydrocodone by LC-MS/MS. Sample #38 tested negative and had semi-quantitative results within 25% of the cutoff.
Traceability and Value Assignment
ARK Hydrocodone Calibrators and Controls are prepared by volumetric dilution of high purity hydrocodone (certified solution traceable to HPLC) into non-sterile, processed human urine free of hydrocodone. Testing is performed with the ARK Hydrocodone Assay on the Beckman Coulter AU680 automated clinical chemistry analyzer, calibrated with the ARK Hydrocodone Calibrator.
Calibration Curve Stability
A stored calibration curve was effective up to at least 14 days based on supporting data. Calibration curve stability may depend on individual laboratory performance.
{16}------------------------------------------------
807.92 (b)(3): Conclusions from Nonclinical Testing
As summarized above, the ARK Hydrocodone Assay is substantially equivalent to the legally marketed predicate device, DRI Hydrocodone Assay (K150502) for the declared intended use. Substantial equivalence has been demonstrated through a comparison of the intended use and device characteristics when comparing the subject device to the legally marketed predicate. Performance testing was completed to verify that the device functions as intended and that design specifications have been satisfied. The content of the pre-market notification for the DRI Hydrocodone Assay provides evidence that the device is safe and effective for the intended use.
§ 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).