The ETERBIO Fentany]Norfentary] Rapid Test (Colloidal Gold) is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

The test is available as a single panel for FYL, or as a dual panel combining both FYL and NFYL. It provides a preliminary screening result only. For a confirmed analytical outcome, a more specific chemical is required. GC/MS or LC/MS is the preferred confirmatory method.

The ETERBIO Fentanyl Norfentanyl Home Test is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

The test is available as a single panel for FYL, or as a dual panel combining both FYL and NFYL. This test provides only preliminary results. For a confirmed analytical outcome, a more specific chemical method is required. GC/MS or LC/MS is the preferred confirmatory method.

Device Description

Rapid Test ETERBIO Fentanyl/Norfentanyl Gold) and ETERBIO The Fentanyl/Norfentanyl Home Test are immunoassays intended for the qualitative detection of fentanyl and norfentanyl in human urine. Each ETERBIO fentanyl/norfentanyl urine test device consists of a Test Panel and a package insert. Each Test Panel is sealed with sachets of desiccant in an aluminum pouch.

AI/ML Overview

The provided text describes the performance characteristics of the ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold) and ETERBIO Fentanyl/Norfentanyl Home Test, which are immunoassay devices for qualitative detection of Fentanyl and Norfentanyl in human urine.

Here's a breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text:

Acceptance Criteria for Performance (Implicit)

The document primarily details the analytical performance and comparison studies without explicitly stating numeric "acceptance criteria" for precision, specificity, or comparison study concordance rates. However, the reported results demonstrate that the device performs as expected for a qualitative drug test around the specified cutoff levels. For the Lay-User Study, the implicit acceptance criterion is a high percentage of correct results and ease of understanding for lay users.

1. Table of Acceptance Criteria and Reported Device Performance

Performance Characteristic	Acceptance Criteria (Implicit from context)	Reported Device Performance (Fentanyl)	Reported Device Performance (Norfentanyl)
Precision	Consistent results across multiple runs and lots, especially near the cutoff concentration. Optimal: 0% false positives/negatives at concentrations far from cutoff, and a mix of positive/negative results near cutoff as expected for qualitative tests.	* -100% to -50% cut off: 60-/0+ (100% negative) across 3 lots. * -25% cut off: Lot 1: 60-/0+, Lot 2: 59-/1+, Lot 3: 60-/0+. * Custom Cutoff Concentration (not explicitly stated): Lot 1: 27+/33-, Lot 2: 26+/34-, Lot 3: 26+/34-. * +25% to +100% cut off: 60+/0- (100% positive) across 3 lots.	* -100% to -50% cut off: 60-/0+ (100% negative) across 3 lots. * -25% cut off: Lot 1: 58-/2+, Lot 2: 58-/2+, Lot 3: 59-/1+. * Custom Cutoff Concentration (not explicitly stated): Lot 1: 27+/33-, Lot 2: 26+/34-, Lot 3: 26+/34-. * +25% to +100% cut off: 60+/0- (100% positive) across 3 lots.
Stability	Device remains stable and functional for a specified shelf life.	24 months at 36-86°F based on real stability study.	24 months at 36-86°F based on real stability study.
Interference	No interference from common physiological/pathological substances or specified concentrations of other compounds.	No interference observed from a comprehensive list of 60+ compounds at specified concentrations (e.g., Acetaminophen, Ethanol, Albumin, Glucose, etc.)	No interference observed from a comprehensive list of 60+ compounds at specified concentrations (e.g., Acetaminophen, Ethanol, Albumin, Glucose, etc.)
Specificity (Cross-Reactivity)	Minimal to no cross-reactivity with structurally similar but non-target compounds, and non-opioid compounds. Appropriate cross-reactivity with relevant metabolites.	Fentanyl: 100% cross-reactivity with Fentanyl at 1 ng/mL. Cross-reactivity observed with several fentanyl analogues (e.g., Acetyl fentanyl (83.3%), Acrylfentanyl (83.3%), Butyryl fentanyl (62.5%), Furanyl fentanyl (57.1%), Isobutyryl fentanyl (66.7%), Ocfentanil (66.7%), Para-fluoro fentanyl (33.3%), Para-fluorobutyryl fentanyl (33.3%), Valeryl fentanyl (40%)). Low/no cross-reactivity with others.	Norfentanyl: 100% cross-reactivity with Norfentanyl at 5 ng/mL. Cross-reactivity observed with several fentanyl analogues (e.g., (+)-3-cis-methyl fentanyl (50%), Acetyl fentanyl (50%), Acrylfentanyl (50%), Butyryl fentanyl (50%), Furanyl fentanyl (55.6%), Isobutyryl fentanyl (62.5%), Para-fluoro fentanyl (100%)). Low/no cross-reactivity with others.
Effect of Urine Specific Gravity & pH	Consistent results across a range of urine specific gravity and pH levels.	All samples at and above +50% Cut-Offs were positive; all samples at and below -50% Cut-Offs were negative, for specific gravity 1.000-1.035 and pH 4-9.	All samples at and above +50% Cut-Offs were positive; all samples at and below -50% Cut-Offs were negative, for specific gravity 1.000-1.035 and pH 4-9.
Method Comparison (Concordance with LC/MS)	High concordance with results from LC/MS (Liquid Chromatography-Mass Spectrometry), especially for samples far from the cutoff. Some discordance expected near cutoff.	Fentanyl: - High agreement for Low Negative and High Positive samples (100% for most operators). - Discordant results mainly near cutoff (e.g., some LC/MS negatives read as positive, some LC/MS positives read as negative). For example, Operator 1 had 1 negative result with LC/MS result (1.01 ng/mL) and 2 positive results with LC/MS results (0.986 ng/mL, 0.937 ng/mL).	Norfentanyl: - High agreement for Low Negative and High Positive samples (100% for most operators). - Discordant results mainly near cutoff. For example, Operator 1 had 1 negative result with LC/MS result (5.2 ng/mL) and 1 positive result (4.85 ng/mL).
Lay-User Study (Accuracy)	High percentage of correct results by lay users, demonstrating ease of use and interpretability.	Fentanyl: 100% correct for samples at -100%, -75%, -50% cutoff, and +25%, +50%, +75% cutoff. 95% correct at -25% cutoff. Norfentanyl: 100% correct for samples at -100%, -75%, -50%, -25% cutoff, and +25%, +50%, +75% cutoff.	Fentanyl: 100% correct for samples at -100%, -75%, -50% cutoff, and +25%, +50%, +75% cutoff. 95% correct at -25% cutoff. Norfentanyl: 100% correct for samples at -100%, -75%, -50%, -25% cutoff, and +25%, +50%, +75% cutoff.
Lay-User Study (Usability)	Instructions are easily understood and followed by lay users.	All lay users indicated that the device instructions can be easily followed. Flesch-Kincaid reading grade level < 7.	All lay users indicated that the device instructions can be easily followed. Flesch-Kincaid reading grade level < 7.

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

Precision Study: For each drug (Fentanyl, Norfentanyl) and each of the 9 concentration levels, 6 tests were performed per day per device lot for 10 days, across 3 lots.
- Total Fentanyl Precision Samples: 9 concentrations * 6 tests/day * 10 days * 3 lots = 1620 tests.
- Total Norfentanyl Precision Samples: 9 concentrations * 6 tests/day * 10 days * 3 lots = 1620 tests.
Interference Study: Not explicitly stated, but implies multiple tests for each of the many interfering substances across three batches of each device.
Specificity Study: Not explicitly stated, but implied tests for each of the listed cross-reactants.
Method Comparison Studies: 80 "unaltered clinical samples" (40 negative, 40 positive) were tested for each drug.
- Total Method Comparison Samples (Fentanyl): 80 samples.
- Total Method Comparison Samples (Norfentanyl): 80 samples.
Lay-User Study: 140 lay persons participated. Urine samples were prepared at 7 different concentrations for each drug, with 20 samples per concentration.
- Total Lay-User Samples (Fentanyl): 7 concentrations * 20 samples/concentration = 140 samples.
- Total Lay-User Samples (Norfentanyl): 7 concentrations * 20 samples/concentration = 140 samples.
Data Provenance: The text does not explicitly state the country of origin for the clinical samples. The studies are described retrospectively in the 510(k) summary, meaning data was collected prior to submission.
- For the Method Comparison Study, samples were "unaltered clinical samples," suggesting they were derived from a patient population.
- For the Precision and Lay-User studies, samples were prepared by spiking fentanyl/norfentanyl into negative urine samples/drug-free pooled urine specimens.

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

Precision, Interference, Specificity, Effect of Urine Specific Gravity & pH Studies:
- Ground truth for these analytical studies was established by preparing samples with known concentrations of drug, verified by LC/MS (Liquid Chromatography-Mass Spectrometry). This is a highly accurate analytical chemistry method. The experts here are implicitly the analytical chemists performing and verifying the LC/MS measurements. The number of such specialists is not specified, but LC/MS is an objective, quantitative method, rather than expert interpretation.
Method Comparison Studies:
- The ground truth was established using LC/MS results. This is an objective analytical standard.
- The "Comparison Studies" section mentions that "three different operators" performed the comparison tests. However, these operators are performing the rapid tests and comparing them to the LC/MS ground truth, they are not establishing the ground truth themselves. The qualifications of these rapid test operators are not specified.
Lay-User Study:
- The ground truth for the prepared samples was established by LC/MS (Liquid Chromatography-Mass Spectrometry).
- Lay users interpreted the results of the rapid test, and their interpretation was compared against the LC/MS ground truth. The "experts" here established the sample concentrations via LC/MS.

4. Adjudication Method for the Test Set

Precision, Interference, Specificity, Effect of Urine Specific Gravity & pH, and Lay-User Studies: Ground truth was established by LC/MS. No human adjudication method (e.g., 2+1, 3+1) is described or needed for these analytical studies where the ground truth is objectively measured by a gold standard (LC/MS).
Method Comparison Studies: The rapid test results from the three operators were directly compared to the LC/MS results. The "Discordant Results" tables list specific samples where the rapid test (by a particular operator or multiple operators) differed from the LC/MS result. There is no explicit human adjudication process described for the rapid test results against the LC/MS ground truth; it's a direct comparison.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

No MRMC comparative effectiveness study was done. This device is a rapid diagnostic test (an immunoassay), not an AI-based imaging diagnostic or a device that assists human "readers" (like radiologists). Therefore, the concept of human readers improving with AI assistance does not apply here. The "operators" in the comparison study are simply performing the rapid test according to its instructions.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was done

This question is not applicable in the context of this device. The ETERBIO Fentanyl/Norfentanyl Rapid Test is a physical immunoassay device where human users (either laboratory personnel or lay users for the home test) directly interpret a visible line. It is not an algorithm that produces a standalone output without human interaction.

7. The Type of Ground Truth Used

The primary ground truth used for performance evaluation (Precision, Interference, Specificity, Method Comparison, Lay-User Studies) was LC/MS (Liquid Chromatography-Mass Spectrometry), which is an objective chemical analysis method considered the gold standard for confirming drug concentrations in urine.
For the Lay-User Study, the ground truth of the samples was LC/MS, and the accuracy of lay user interpretation was assessed against this LC/MS truth.

8. The Sample Size for the Training Set

This device is an immunoassay, not a machine learning or AI-based device that requires a "training set" in the computational sense. Therefore, this concept does not apply to the ETERBIO Fentanyl/Norfentanyl Rapid Test. Its "training" is inherent in its chemical design and manufacturing process.

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

As stated above, the device does not have a "training set" in the context of machine learning. Its analytical performance relies on validated chemical reactions and manufacturing consistency.

Summary

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November 20, 2024

Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

Eterbio, Inc. % Jenny Xia Director LSI International Inc 504 E Diamond Ave Suite H Gaithersburg, Maryland 20877

Re: K243064

Trade/Device Name: ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold); ETERBIO Fentanyl/Norfentanyl Home Test Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: Class II Product Code: NGL Dated: September 26, 2024 Received: September 27, 2024

Dear Jenny Xia:

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.

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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).

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.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

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-regulatory

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assistance/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,

Image /page/2/Picture/3 description: The image shows a digital signature. The signature is for Joseph A. Kotarek -S. The date of the signature is 2024.11.20, and the time is 09:04:19 -05'00'.

Joseph Kotarek, Ph.D. 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) K243064

Device Name

ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold) ETERBIO Fentanyl/Norfentanyl Home Test

Indications for Use (Describe)

The ETERBIO Fentany]Norfentary] Rapid Test (Colloidal Gold) is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

The ETERBIO Fentanyl Norfentanyl Home Test is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

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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510(k) SUMMARY

K243064

1. Date: October 21, 2024 2. Submitter: Eterbio, Inc. 420, No. 1316, Caixia Street, Zhuhai, Guangdong China 3. Contact person: Jenny Xia LSI International Inc. 504E Diamond Ave., Suite H Gaithersburg, MD 20877 Telephone: 240-505-7880 Email: jxia@lsi-consulting.org 4. Device Names: ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold) ETERBIO Fentanyl/Norfentanyl Home Test

Product Code	Classification	Regulation Section	Panel
NGL	II	21 CFR § 862.3650Opiate Test System	Toxicology (91)

1. Predicate Devices:
  Hightop® Home Use Fentanyl/Norfentanyl Urine Rapid Test Panel (K241969)
1. Indications for Use
  The ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold) is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

The test is available as a single panel for FYL or NFYL, or as a dual panel combining both FYL and NFYL. It provides a preliminary screening result only. For a confirmed analytical outcome, a more specific chemical method is required. GC/MS or LC/MS is the preferred confirmatory method.

The ETERBIO Fentanyl Norfentanyl Home Test is an immunoassay intended for the qualitative detection of Fentanyl (FTY) /Norfentanyl (NFTY) in human urine:

Drug (Identifier)	Calibrator	Cut-off Level
Fentanyl (FYL)	Fentanyl	1ng/mL
Norfentanyl (NFYL)	Norfentanyl	5ng/mL

The test is available as a single panel for FYL or NFYL, or as a dual panel combining both FYL and NFYL. This test provides only preliminary results. For a confirmed analytical outcome, a more specific chemical method is required. GC/MS or LC/MS is the preferred confirmatory method.

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1. Device Description
  Rapid Test ETERBIO Fentanyl/Norfentanyl Gold) and ETERBIO The Fentanyl/Norfentanyl Home Test are immunoassays intended for the qualitative detection of fentanyl and norfentanyl in human urine. Each ETERBIO fentanyl/norfentanyl urine test device consists of a Test Panel and a package insert. Each Test Panel is sealed with sachets of desiccant in an aluminum pouch.

Substantial Equivalence Information A summary comparison of features of the ETERBIO Fentanyl Test and the predicate devices is provided in following table.



Item	Device	Predicate – K241969
Indication(s)for Use	For the qualitative determination offentanyl in human urine.	Same
Calibrator and Cut-OffValues	Fentanyl (FTY)1 ng/mlNorfentanyl (NFTY)5 ng/ml	Same
Methodology	Competitive binding, lateral flowimmunochromatographic assays basedon the principle of antigen antibodyimmunochemistry.	Same
Type of Test	Qualitative	Same
Specimen Type	Human Urine	Same
Intended Use	For OTC use	Same
Configurations	Panel	Same
Storage	2-30°C	Same

1. Test Principle
  ETERBIO The ETERBIO Fentanyl/Norfentanyl Rapid Test (Colloidal Gold) and Fentanyl/Norfentanyl Home Test are immunoassays based on the principle of competitive binding.

During testing, a urine specimen migrates upward by capillary action. Each target drug, if present in the urine specimen below its cutoff, will not saturate the binding sites of antibodycoated particles in the test device. The antibody-coated particles will then be captured by immobilized drug conjugate and a visible colored line will show up in the test line region. The colored line will not form in the test line region if the drug level exceeds the cutoff because it will saturate all the binding sites of anti-drug antibodies.

To serve as a procedural control, a colored line will always appear at the control line region indicating that proper volume of specimen has been added and membrane wicking has occurred.

1. Performance Characteristics

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1. Analytical Performance

a. Precision

Precision studies were carried out for samples with concentrations of -100% cut off. -75% cut off, -50% cut off, -25% cut off, cut off, +25% cut off, +50% cut off, +75% cut off and +100% cut off for each target drug. These samples were prepared by spiking fentanyl or norfentanyl in negative urine samples. Each fentanyl or norfentanyl concentration was confirmed by LC/MS. All sample aliquots were blindly labeled by the person who prepared the samples and didn't take part in the sample testing. For each concentration, tests were performed six tests per day per device lot for 10 days in a randomized order.

Fentanyl

LotNumber	-100%cut off	-75%cut off	-50%cut off	-25%cutoff	+25%cut off	+50%cut off	+75%cut off	+100%cut off
Lot 1	60-/0+	60-/0+	60-/0+	60-/0+	27+/33-	60+/0-	60+/0-	60+/0-
Lot 2	60-/0+	60-/0+	60-/0+	59-/1+	26+/34-	60+/0-	60+/0-	60+/0-
Lot 3	60-/0+	60-/0+	60-/0+	60-/0+	26+/34-	60+/0-	60+/0-	60+/0-

Norfentanyl

LotNumber	-100%cut off	-75%cut off	-50%cut off	-25%cutoff	cut off	+25%cut off	+50%cut off	+75%cut off	+100%cut off
Lot 1	60-/0+	60-/0+	60-/0+	58-/2+	27+/33-	60+/0-	60+/0-	60+/0-	60+/0-
Lot 2	60-/0+	60-/0+	60-/0+	58-/2+	26+/34-	60+/0-	60+/0-	60+/0-	60+/0-
Lot 3	60-/0+	60-/0+	60-/0+	59-/1+	26+/34-	60+/0-	60+/0-	60+/0-	60+/0-

c. Stability

The devices are stable at 36-86F for 24 months based on the real stability study.

d. Interference

Potential interfering substances found in human urine of physiological or pathological conditions were added to drug-free urine and target drug fentanyl urine with concentrations at 50% below and 50% above Cut-Off levels. These urine samples were tested using three batches of each device. Compounds that showed no interference at a concentration of 100µg/mL or specified concentrations are summarized in the following tables.

Acetaminophen	Creatinine	Ketamine	Perphenazine
Acetone (1000mg/dL)	Cyclobenzaprine	Ketoprofen	Phencyclidine
Acetophenetidin	Deoxycorticosterone	Labetalol	Phenelzine
Acetylsalicylic acid	Desipramine	Lidocaine	Phenobarbital
Albumin (100mg/dL)	Dextromethorphan	Loperamide	Prednisone
Albuterol	Diclofenac	Maprotiline	Propoxyphene
Aminopyrine	Diflunisal	Meperidine	Propranolol
Amitriptyline	Digoxin	Meprobamate	Pseudoephedrine
Amobarbital	Diphenhydramine	Methapyrilene	Quinine
Amoxicillin	DL-Tryptophan	Methaqualone	Ranitidine
Ampicillin	DL-Tyrosine	Methoxyphenamine	Riboflavin (10mg/dL)
Apomorphine	Doxepin	Metronidazole	Salicylic acid
Ascorbic acid	Ecgonine methyl ester	N-Acetylprocainamide	Secobarbital
Aspartame	Ephedrine	NaCl (4000mg/dL)	Serotonin(5-Hydroxytyramine)
Atropine	Erythromycin	Nalidixic acid	Sulfamethazine
Benzilic acid	Ethanol (1%)	Naloxone	Sulindac
Benzoic acid	Fenoprofen	Naltrexone	Tetrahydrocortisone 3-(β-Dglucuronide)
Benzoylecgonine	Fluphenazine	Naproxen	Tetrahydrocortisone 3-acetate
Bilirubin	Furosemide	Niacinamide	Tetrahydrozoline
Boric Acid (1%)	Galactose (10mg/dL)	Nicotine	Thiamine

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Bupropion	Gamma Globulin (500mg/dL)	Nifedipine	Thioridazine
Caffeine	Gentisic acid	Norethindrone	Triamterene
Carbamazepine	Glucose (3000mg/dL)	Nortriptyline	Trifluoperazine
Chloral hydrate	Hemoglobin	Noscapine	Trimethoprim
Chloramphenicol	Hydralazine	O-Hydroxyhippuric acid	Tyramine
Chlorothiazide	Hydrochlorothiazide	Octopamine	Urea (2000mg/dL)
Chlorpromazine	Hydrocortisone	Oxalic acid (100mg/dL)	Uric acid
Cholesterol	Hydroxytyramine	Oxazepam	Valproic acid (250µg/mL)
Clomipramine	Ibuprofen	Oxolinic acid	Venlafaxine
Clonidine	Imipramine	Oxymetazoline	Verapamil
Cortisone	Isoproterenol	Papaverine	Zomepirac
Cotinine	Isoxsuprine (10µg/mL)	Penicillin G	β-Estradiol
α-Estradiol	Estrone	7-Aminonitrazepam	Fenfluramine
Fenofibrate	Amlodipine besylate	Phentermine	Phenylethylamine
Fotemustine	Promazine	Promethazine	Gemfibrozil
Aspirin	Gentisic acid	Pyridoxine	Baclofen
Guaiacolglyceryl ether	Pyrilamine	Pyrogallol	Benzocaine
Hexobarbital	Quinidine	Quinolinic Acid	Benzylpiperiazine
Hydroxybutyric Acid	Bromo-2,5,Dimethoxyphenethylamine	Sodium Azide	Carisoprodol
Cetirizine	Lamotrigine	Tetracycline	Lisinopril
Chlordiazepoxide	Loratidine	Chlorpheniramine	Trifluoromethylphenyl-piperazine
Clofibrate	Tryptamine	Metoprolol	Creatine Hydrate
N-desmethyl Tapentadol	γ-Cyclodextrin	Zolpidem	Cyproheptadine
Demoxepam	7-Aminoflunitrazepam	Metformin	Norpseudoephedrine
Nicotinic Acid	Oxazepam Glucuronide	Lorazepam Glucuronide	LSD
Dimethyl-aminoantipyrine	THC	L-thyroxine	Diphenylhydantoin
Methylphenidate	Norpropoxyphene

e.Specificity

To test specificity, drug metabolites and other components that are likely to interfere in urine samples were tested using three batches of device. The lowest concentration that caused a positive result for each compound are listed below.

Fentanyl (Cutoff=1ng/mL)	Minimumconcentrationrequired to obtain apositive result(ng/mL)	% Cross-Reactivity
(±) β-hydroxythiofentanyl	2.8	35.7%
(±)-3-cis-methyl fentanyl	5	20%
4-Fluoro-isobutyrylfentanyl	3	33.3%
9-Hydroxy Risperidone	10000	0.01%
Acetyl fentanyl	1.2	83.3%
Acetyl norfentanyl	10000	0.01%
Acrylfentanyl	1.2	83.3%
Alfentanil	100000	0.001%
Butyryl fentanyl	1.6	62.5%
Carfentanil	500	0.20%
Despropionyl fentanyl (4-ANPP)	50000	0.002%
Fentanyl	1	100%
Furanyl fentanyl	1.75	57.1%
Isobutyryl fentanyl	1.5	66.7%
Labetalol Hydrochloride	100000	0.001%
MT-45	10000	0.01%
Norcarfentanil	10000	0.01%

{8}------------------------------------------------

Norfentanyl	5000	0.02%
Ocfentanil	1.5	66.7%
Para-fluoro fentanyl	3	33.3%
Para-fluorobutyryl fentanyl	3	33.3%
Remifentanil	10000	0.01%
Risperidone	1000	0.1%
Sufentanil	625	0.16%
Thienyl Fentnayl	1000	0.1%
Trans-d, I 3-Methylfentanyl	1000	0.1%
Trazodone	1000	0.1%
U-47700	100000	0.001%
Valeryl fentanyl	2.5	40 %
ω- 1-Hydroxyfentanyl	20000	0.005%

Norfentanyl (Cutoff=5ng/mL)	Minimumconcentrationrequired to obtain apositive result(ng/mL)	% Cross-Reactivity
(+) β-hydroxythiofentanyl	7	71.4%
(+)-3-cis-methyl fentanyl	10	50%
4-Fluoro-isobutyrylfentanyl	25	20%
9-Hydroxy Risperidone	10000	0.05%
Acetyl fentanyl	10	50%
Acetyl norfentanyl	200	2.5%
Acry lfentanyl	10	50%
Alfentanil	>100000/mL	<0.001%
Butyryl fentanyl	10	50%
Carfentanil	>100000/mL	<0.001%
Despropionyl fentanyl (4-ANPP)	>100000/mL	<0.001%
Fentanyl	15	33.3%
Furanyl fentanyl	9	55.6%
Isobutyryl fentanyl	8	62.5%
Labetalol Hydrochloride	20	25%
MT-45	100000	0.005%
Norcarfentanil	5000	0.1%
Norfentanyl	5	100%
Ocfentanil	15	33.3%
Para-fluoro fentanyl	5	100%
Para-fluorobutyryl fentanyl	40	12.5%
Remifentanil	>100000/mL	<0.001%
Risperidone	1000	0.5%
Sufentanil	>100000/mL	<0.001%
Thienyl Fentnayl	50	10%
Trans-d, I 3-Methylfentanyl	50	10%
Trazodone	>100000/mL	<0.001%

{9}------------------------------------------------

U-47700	100000	0.005%
Valeryl fentanyl	100	5%
ω- 1-Hydroxyfentanyl	500	1%

The following opioids compounds were tested at a concentration of 100ug/mL. Negative results were obtained for all these compounds. There is no cross-reactivity for these compounds using the ETERBIO Device.

6-Acetyl morphine	Naloxone
Amphetamine	Naltrexone
Buprenorphine	Norbuprenorphine
Buprenorphineglucuronide	Norcodeine
Codeine	Norketamine
Dextromethorphan	Normeperidine
Dihydrocodeine	Normorphine
EDDP	Noroxycodone
EMDP	Oxycodone
Fluoxetine	Oxymorphone
Heroin	Pentazocine (Talwin)
Hydrocodone	Pipamperone
Hydromorphone	Tapentadol
Ketamine	Thioridazine
Levorphanol	Tilidine
Meperidine	Tramadol
Methadone	Tramadol-O-Desmethyl
Morphine	Tramadol-N-Desmethyl
Morphine-3-glucuronide

f. Effect of Urine Specific Gravity and Urine pH

To investigate the effect of urine specific gravity and urine pH, urine samples, with 1.000 to 1.035 specific gravity or urine samples with pH 4 to 9 were spiked with targets fentanyl and norfentanyl at 50% below and 50% above Cut-Off levels. These samples were tested using three lots of device. Results were all positive for samples at and above +50% Cut-Offs and all negative for samples at and below -50% Cut-Offs.

2. Comparison Studies

Method comparison studies for the ETERBIO Fentanyl Test were performed by three different operators. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples. The samples were blind labeled and compared to LC/MS results are presented in the tables below.

			LowNegative byLC/MS(less than-50%)	Near CutoffNegative byLC/MS(Between-50% andcutoff)	Near CutoffPositive byLC/MS(Between thecutoff and+50%)	High Positiveby LC/MS(greater than+50%)
Operator1	Positive	0	0	2	19	20
	Negative	8	16	14	1	0

Fentanyl

{10}------------------------------------------------

Operator		Positive	0	0	1	18	20
2	Negative	8	16	15	2	0
Operator		Positive	0	0	2	20	20
3	Negative	8	16	14	0	0

Discordant Results

Operator	Sample ID	LC/MS Result (ng/mL)	Rapid Test Result
Operator 1	MF046	0.986	+
Operator 1& Operator 3	MF079	0.937	+
Operator 2	MF062	0.925	+
Operator 3	MF058	0.914	+
Operator 2	MF016	1.02	-
Operator 1	MF060	1.01	-
Operator 2	MF063	1.09	-

Norfentanyl

		Negative	Low Negative byLC/MS(less than -50%)	Near Cutoff Negative byLC/MS(Between -50% andcutoff)	Near Cutoff Positive byLC/MS(Between thecutoff and +50%)	High Positiveby LC/MS(greater than+50%)
Operator1	Positive	0	0	1	19	20
	Negative	8	17	14	1	0
Operator2	Positive	0	0	1	19	20
	Negative	8	17	14	1	0
Operator3	Positive	0	0	1	20	20
	Negative	8	17	14	0	0

Discordant Results

Operator	Sample ID	LC/MS Result (ng/mL)	Rapid Test Result
Operator 1	MC083	5.2	-
Operator 2	MC019	5.25	-
Operator 1 & Operator 2	MC048	4.85	+
Operator 3	MC009	4.68	+

3. Lay-user study

A lay user study was performed at three testing sites with 140 lay persons. They had diverse educational and professional backgrounds and ranged in age from 18 to >50 years. Urine samples were prepared at the following concentrations; -100%, +/-50%, +/-50%, +/-25% of the cut-offs by spiking drug(s) into drug free-pooled urine specimens. The concentrations of the samples were confirmed by LC/MS. Each sample was aliquoted into individual containers, blind-labeled and randomized. Each participant was provided with the package insert, 1 blind labeled sample and a device. The results are summarized below:

% of Cutoff	Number	Fentanyl Concentration	Lay person results	The
-------------	--------	------------------------	--------------------	-----

{11}------------------------------------------------

	ofsamples	by LC/MS(ng/mL)	No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100.0%
-75% Cutoff	20	0.25	0	20	100.0%
-50% Cutoff	20	0.52	0	20	100.0%
-25% Cutoff	20	0.75	1	19	95.0%
+25% Cutoff	20	1.30	20	0	100.0%
+50% Cutoff	20	1.55	20	0	100.0%
+75% Cutoff	20	1.65	20	0	100.0%

% of Cutoff	Number of samples	Norfentanyl Concentration by LC/MS (ng/mL)	Lay person results		The percentage of correct results (%)
-100% Cutoff	20	0	0	20	100.0%
-75% Cutoff	20	1.27	0	20	100.0%
-50% Cutoff	20	2.54	0	20	100.0%
-25% Cutoff	20	3.78	0	20	100.0%
+25% Cutoff	20	6.55	20	0	100.0%
+50% Cutoff	20	7.85	20	0	100.0%
+75% Cutoff	20	8.95	20	0	100.0%

Lay-users were also given surveys on the ease of understanding the package insert instructions. All lay users indicated that the device instructions can be easily followed. A Flesch-Kincaid reading analysis was performed on the package insert and the score revealed a reading grade level of less than 7.

1. Clinical Studies
  Not applicable.
1. Conclusion
  Based on the test principle and acceptable performance characteristics including precision, cut-off, interference, specificity, method comparison and Lay-user studies of the devices, it's concluded a substantial equivalence decision.

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).