BioSieve™ Fentanyl FIA Test Kit is a fluorescence immunoassay (FIA) for the qualitative determination of fentanyl in human urine at a cutoff concentration of 1.0 ng/mL. The assay is intended for use with BioSieve™ ToxiSmart FIA Reader.

It is for in vitro diagnostic use only. It is intended for prescription use.

The tests provide only preliminary results. A more specific alternative chemical must be used in order 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.

Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.

BioSieve™ ToxiSmart FIA Reader is a portable fluorescence instrument for in vitro diagnostic use only. The Reader is designed to perform in vitro diagnostic tests on urine specimens. This Reader can be used in a laboratory or in a point-of-care setting.

Device Description

This test uses a lateral flow design with location-dependent lines and zones. BioSieve™ ToxiSmart FIA Reader scans the test strip and displays results. The sample is added to the sample well of the test card, and the sample is drawn by capillary action into and through the fluorescent labeled pad, through the nitrocellulose strip and into the adsorption pad. Within the fluorescent labeled pad, the specimen comes into contact with antibodies conjugated with fluorescent microspheres. During this interaction, if the amount of fentanyl antigen in the sample is greater than or equal to the cutoff value, the antigen in the sample and the fluorescence-labeled antibody bind to the FTY antigenantibody complex when the sample passes through a pad of fluorescence-microbead-labeled antibody conjugate. As the sample flows and reaches the FTY antigen coated by the T-line of nitrocellulose membrane, the FTY antigen coated by the T-line antigen in the sample competitively bind the FTY antibody labeled with fluorescence, then the T-line captures fluorescence signal is weaker than the cutoff fluorescence signal. When the samples do not contain fentanyl antigen or levels below the cutoff value, as the sample flow, fluorescent microsphere labeled antibody to nitrocellulose membrane T line captures fluorescent signal is stronger than the cutoff fluorescence signal. Whether or not FTY antigen was present in the sample, the rabbit IgG fluorescent microsphere conjugate not bound to the test line continued to flow with the rest of the sample and soon encountered a control line composed of goatanti-rabbit IgG. The position of C-line will accumulate fluorescence signal. The C-line control area was scanned to confirm that adequate sample flow had occurred. High resolution, narrow band SMD LED was used as light source in the Immunofluorescence Analyzer. The central wavelength of the excitation spectrum is 365nm. The central response wavelength is 610nm.

AI/ML Overview

The provided text describes the acceptance criteria and the study that proves the device meets those criteria for the BioSieve™ Fentanyl FIA Test Kit and BioSieve™ ToxiSmart FIA Reader.

Here's an analysis of the requested information:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are not explicitly stated as distinct pass/fail thresholds in a table format. However, the performance characteristics tested and summarized imply the criteria for acceptable performance. The study aims to demonstrate that the device performs as expected for a qualitative immunoassay for fentanyl in urine, particularly around the 1.0 ng/mL cutoff.

The reported device performance is presented in several sections:

Performance Metric	Implied Acceptance Criteria (Based on typical immunoassay requirements)	Reported Device Performance
Precision	High agreement for samples well below and well above cutoff; reasonable agreement near cutoff.	For -100%, -75%, -50% cutoff, all 60 tests for each lot were negative (60-/0+). For +25%, +50%, +75%, +100% cutoff, all 60 tests for each lot were positive (60+/0-). For -25% cutoff, 58-59 negative and 1-2 positive out of 60. For cutoff (1.0 ng/mL), 28-29 negative and 31-32 positive out of 60.
Stability	Device remains effective under specified storage conditions for its shelf life.	Stable at 2-30°C for 24 months based on accelerated stability study; real-time ongoing.
Interference	No significant interference from common substances found in urine.	Numerous compounds (e.g., Acetaminophen, Ethanol, Glucose) at specific concentrations showed no interference.
Specificity (Cross-Reactivity)	Low or no cross-reactivity with other related compounds or metabolites not intended to be detected, especially at clinically relevant concentrations.	Lists various fentanyl-related compounds and their cross-reactivity percentages (e.g., Acetyl fentanyl 83.33%, Carfentanil 0.20%). Many opiate compounds showed no cross-reactivity at 100 µg/mL.
Effect of Urine Specific Gravity and pH	Performance should be robust across a physiological range of urine specific gravity and pH.	Samples spiked at -50% and +50% Cut-Off levels across specific gravity 1.000-1.035 and pH 4-9 showed expected negative and positive results respectively.
Method Comparison (Clinical Samples)	High concordance with a reference method (LC-MS/MS), especially for samples far from the cutoff. Acceptable levels of discordance near the cutoff.	Site 1: 0 False Positives (FP) from negative, 0 FP from low negative, 4 FP from near cutoff negative. 1 False Negative (FN) from near cutoff positive, 0 FN from high positive. Site 2: 0 FP from negative, 0 FP from low negative, 3 FP from near cutoff negative. 2 FN from near cutoff positive, 0 FN from high positive. Site 3: 0 FP from negative, 0 FP from low negative, 2 FP from near cutoff negative. 1 FN from near cutoff positive, 0 FN from high positive. Discordant Results: Specific sample numbers and their LC-MS/MS vs. BioSieve results are listed, showing cases where the device gave a positive result for a true negative near the cutoff, and a negative result for a true positive near the cutoff.

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

Precision Study Test Set: For each of 9 concentrations (e.g., -100% cutoff, cutoff, +100% cutoff), 6 runs per day for 10 days per device lot were performed. With 3 lots, this suggests: 9 concentrations * 6 tests/run * 10 days * 3 lots = 1620 individual tests (results reported as sums across these).
Interference Study Test Set: Numerous interfering substances were tested with 3 batches of each device for drug-free urine and target fentanyl urine at -50% and +50% Cut-Off levels. The exact number of tests per substance is not specified, but it implies a substantial number.
Specificity (Cross-Reactivity) Test Set: Various drug metabolites and opioid compounds were tested using three batches of device. The number of individual tests per compound is not explicitly stated.
Effect of Urine Specific Gravity and pH Test Set: Urine samples across the specified ranges were spiked at -50% and +50% Cut-Off. Tested by three different operators per lot of device, with a total of three lots. The exact number of individual tests is not specified.
Method Comparison Test Set: 80 unaltered clinical samples (40 negative and 40 positive based on an internal classification) were used per site across three different testing sites. This totals 240 clinical samples.
Data Provenance: The document does not explicitly state the country of origin for the samples. It mentions "unaltered clinical samples" for the method comparison study, implying they were retrospective if they were already collected clinical samples. The precision study samples were "prepared by spiking fentanyl in negative samples," indicating these were artificially contrived samples, not naturally occurring clinical specimens, and were conducted in a controlled lab setting.

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

For the Precision Study, "Each fentanyl concentration was confirmed by LC/MS-MS." LC/MS-MS (Liquid Chromatography/Mass Spectrometry) is a highly accurate analytical chemistry method, typically considered a gold standard for quantifying substances in biological samples. This method itself provides the "ground truth" and does not typically involve human "experts" in the sense of clinicians or radiologists reading results.
For the Method Comparison Study, "The samples were blind labeled and compared to LC-MS/MS results." Again, LC-MS/MS is the ground truth. No human experts (e.g., radiologists) were used to establish the ground truth; it was established by an analytical instrument method.
No information is provided about the qualifications of the operators who performed the tests on the BioSieve™ device at the three sites, nor is there mention of a panel of experts for subjective adjudication.

4. Adjudication Method for the Test Set

None in the context of human expert adjudication. The ground truth for quantitative accuracy and comparative performance was established by LC-MS/MS, a definitive analytical method, not by human consensus or adjudication. The BioSieve™ device provides a qualitative result (positive/negative), which is then compared against the quantitative LC-MS/MS value relative to the 1.0 ng/mL cutoff.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC study was not done. This device is a quantitative/qualitative immunoassay read by an instrument (BioSieve™ ToxiSmart FIA Reader), not interpreted by human readers in the same way an imaging AI would be. The "operators" in the method comparison study are likely technicians performing the test, not interpreting complex outputs. Therefore, a study on human reader improvement with AI assistance is not applicable.

6. Standalone (Algorithm Only Without Human-in-the-Loop Performance) Study

Yes, in essence. The BioSieve™ ToxiSmart FIA Reader is an automated instrument that reads the test strip and displays results. The results presented for precision, interference, specificity, and comparison studies demonstrate the standalone performance of the BioSieve™ Fentanyl FIA Test Kit and the BioSieve™ ToxiSmart FIA Reader system. The device produces a qualitative result directly from the sample, without human interpretation of the signal beyond observing the final positive/negative display.

7. Type of Ground Truth Used

LC-MS/MS (Liquid Chromatography/Mass Spectrometry): This highly accurate analytical method was used to confirm fentanyl concentrations in precision samples and as the reference method for clinical samples in the method comparison study. It provides objective, quantitative biochemical data, which is then translated to the qualitative positive/negative ground truth based on the 1.0 ng/mL cutoff.

8. Sample Size for the Training Set

Not specified. The document describes performance validation studies for regulatory submission (510(k)). It does not provide details about a specific "training set" for an AI algorithm because the BioSieve™ ToxiSmart FIA Reader is not described as an AI-powered device in the typical sense of a deep learning model that requires a labeled training dataset. It's a fluorescence immunoassay reader. The "development modes" refer to how the test is run (Standard vs. Quick), not algorithmic training.

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

Not applicable. As noted above, the device is an immunoassay system and reader, not explicitly an AI/machine learning algorithm requiring a training set with established ground truth in the context of typical AI development. The document describes analytical validation, not algorithmic training.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services seal on the left and the FDA acronym along with the full name of the agency on the right. The FDA acronym is in a blue square, followed by the words "U.S. Food & Drug Administration" in blue.

VivaChek Biotech (Hangzhou) Co., Ltd % Joe Shia Director LSI International Inc 504 E Diamond Ave., Suite H Gaithersburg, Maryland 20877

Re: K240124

Trade/Device Name: BioSieve™ Fentanyl FIA Test Kit; BioSieve™ ToxiSmart FIA Reader Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: Class II Product Code: DJG, KHO Dated: May 10, 2024 Received: May 10, 2024

Dear Joe Shia:

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 Joseph A. Kotarek -S
Date: 2024.06.14
14:08:09 -04'00'

Joseph Kotarek 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

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Indications for Use

510(k) Number (if known) K240124

Device Name BioSieve™ Fentanyl FIA Test Kit BioSieve™ ToxiSmart FIA Reader

Indications for Use (Describe)

It is for in vitro diagnostic use only. It is intended for prescription use.

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)	Prescription Use (Part 21 CFR 801 Subpart D)	Prescription Use (Part 21 CFR 801 Subpart D)		Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)	Over-The-Counter Use (21 CFR 801 Subpart C)		Over-The-Counter Use (21 CFR 801 Subpart C)
Prescription Use (Part 21 CFR 801 Subpart D)	Prescription Use (Part 21 CFR 801 Subpart D)		Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)	Over-The-Counter Use (21 CFR 801 Subpart C)		Over-The-Counter Use (21 CFR 801 Subpart C)
Prescription Use (Part 21 CFR 801 Subpart D)		Prescription Use (Part 21 CFR 801 Subpart D)
	Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)		Over-The-Counter Use (21 CFR 801 Subpart C)
	Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) SUMMAR Y

K240124

1. Date:	June 12, 2024
2. Submitter:	VivaChek Biotech (Hangzhou) Co., Ltd.Level 2, Block 2, 146 East ChaofengRd. Yuhang Economy DevelopmentZone Hangzhou 311100 ZhejiangP.R. China
3. Contact person:	Joe ShiaLSI International Inc.504E Diamond Ave., Suite IGaithersburg, MD 20877Telephone: 240-505-7880Email: shiajl@yahoo.com
4. Device Names:	BioSieve™ Fentanyl FIA Test Kit

Classification: Class 2
Product Code	Classification	Regulation Section	Panel
DJG	II	21 CFR § 862.3650Opiate Test System	Toxicology (91)
KHO	I	21 CFR § 862.2560Fluorometer for clinical use	Clinical Chemistry

BioSieve™ ToxiSmart FIA Reader

5. Predicate Devices:

Superbio Fentanyl Urine Detection Kit, K220046; Superbio Immunofluorescence Analyzer EASY-11, K220046

1. Indications for Use
  BioSieve™ Fentanyl FIA Test Kit is a fluorescence immunoassay (FIA) for the qualitative determination of fentanyl in human urine at a cutoff concentration of 1.0 ng/mL. The assay is intended for use with BioSieve™ ToxiSmart FIA Reader.

It is for in vitro diagnostic use only. It is intended for prescription use.

Clinical consideration and professional judgment should be exercised with any drug test result. particularly when the preliminary test result is positive.

1. Device Description
  This test uses a lateral flow design with location-dependent lines and zones. BioSieve™ ToxiSmart FIA Reader scans the test strip and displays results. The sample is added to the sample well of the

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test card, and the sample is drawn by capillary action into and through the fluorescent labeled pad, through the nitrocellulose strip and into the adsorption pad. Within the fluorescent labeled pad, the specimen comes into contact with antibodies conjugated with fluorescent microspheres. During this interaction, if the amount of fentanyl antigen in the sample is greater than or equal to the cutoff value, the antigen in the sample and the fluorescence-labeled antibody bind to the FTY antigenantibody complex when the sample passes through a pad of fluorescence-microbead-labeled antibody conjugate. As the sample flows and reaches the FTY antigen coated by the T-line of nitrocellulose membrane, the FTY antigen coated by the T-line antigen in the sample competitively bind the FTY antibody labeled with fluorescence, then the T-line captures fluorescence signal is weaker than the cutoff fluorescence signal. When the samples do not contain fentanyl antigen or levels below the cutoff value, as the sample flow, fluorescent microsphere labeled antibody to nitrocellulose membrane T line captures fluorescent signal is stronger than the cutoff fluorescence signal. Whether or not FTY antigen was present in the sample, the rabbit IgG fluorescent microsphere conjugate not bound to the test line continued to flow with the rest of the sample and soon encountered a control line composed of goatanti-rabbit IgG. The position of C-line will accumulate fluorescence signal. The C-line control area was scanned to confirm that adequate sample flow had occurred. High resolution, narrow band SMD LED was used as light source in the Immunofluorescence Analyzer. The central wavelength of the excitation spectrum is 365nm. The central response wavelength is 610nm.

1. Substantial Equivalence Information
  A summary comparison of features of the BioSieve™ Fentanyl FIA Test Kit and the predicate devices is provided in following table.

Item	Device	Predicate - K220046
Indication(s)for Use	For the qualitative determination offentanyl in human urine.	Same
Calibrator and Cut-OffValues	Fentanyl (FTY)1 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 prescription use	Same
Configurations	Test Card (chip on the back)	Test Card (no back chip)
Platform Required	Immunofluorescence Analyzer	Same
Storage	2-30°C	4-30°C
Item	Device	Predicate - K220046
Intended Use/ Indication forUse	Immunofluorescence analyzerdesigned to perform in vitrodiagnostic tests on clinicalspecimens including drug urine	Same
Principles of AssayOperation	Competitive immunofluorescenceimmunoassay	Same
Calibration Check	A Quality control test device is suppliedwith the Reader and used for check theReader optics and calculation systems.	A Quality control card issupplied with Easy-11 andused for check Easy-11 opticsand calculation. systems.
Development Modes	Two basic assay development modes:• Standard test: In standard test, the userinsert the Reader immediately afteradding the sample, and the Reader willdisplay the test result when thecountdown is finished• Quick test: In the quick test, the userneed insert the Reader after the reactiontime is completed, and the Reader willdisplay the test result in a few seconds.	Two basic assay developmentmodes:• Standard test: In standard test,the user immediately insertsTest Cassette into Easy-11 andclick "start test". Easy-11automatically counts the time.• Quick test: Manually timing,then insert the test card intoEasy-11, and click "Start Test".The instrument will read theresults.
User interface	1.54 inch LCD Screen display	8 inch Color LCD touchscreendisplay
Barcode scanner (sample)	Not equipped with a barcode scanner	Same
Assay/instrumentinterface	Drawer	Same
Light Source	LED Light	Same
Power Supply	Powered by a 3.7V lithium-ion batteryTwo charging methods:1.Type C & USB 2 in 1 cable (computercharging)2.Type C & USB 2 in 1 cable with the ACadapter (wall charging)Input: 100-240V~, 50/60Hz, 0.2A Max;Output: 5.0V=, 1.0A	AC100-240V
Dimensions	12.45 cm x 7.25 cm x 4 cm	28 cm x 28 cm x 16 cm

Table 1: Features Comparison of BioSieve™ Fentanyl FIA Test Kit and the Predicate Devices

Table 2: Instrument Similarities and Differences

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9. Test Principle

BioSieve™ Fentanyl FIA Test Kit uses the principle of competitive and fluorescence immunochromatography assay. The nitrocellulose membrane test area (T) of the test strip is correspondingly coated with fentanyl-bovine serum albumin conjugate, and the quality control area (C) is coated with goat anti-rabbit lgG polyclonal antibody. Both Fentanyl monoclonal antibody and rabbit lgG polyclonal antibody labeled with fluorescent microspheres are embedded on the conjugate pad. The labeled antibody will flow forward with the sample, when the urine sample is applied to the sample well of the test device. When the concentration of fentanyl in the sample is higher than or equal to the cut-off of the product, it will compete with the corresponding conjugate coated on the test area (T) to bind to the fluorescently labeled monoclonal antibody, the fluorescence signal rendering of the test line is inhibited and the result is positive; while when the sample does not contain fentanyl or its concentration is lower than the cut-off of the product, the corresponding conjugate on the test line reacts with sufficient fluorescently-labeled monoclonal antibodies, the test line will have fluorescence signal and the result is negative. The quality control area (C) will develop fluorescence signal, which is the criteria for judging whether the test process is normal or not. Signal intensity of fluorescence is detected by BioSieve™ ToxiSmart FIA Reader.

10. Performance Characteristics

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. These samples were prepared by spiking fentanyl in negative samples. Each fentanyl concentration was confirmed by LC/MS-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 runs per day for 10 days per device lot in a randomized order. Each device was read on one BioSieve™ ToxiSmart FIA Reader. The results obtained are summarized in the following tables.

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+	28-/32+	60+/0-	60+/0-	60+/0-	60+/0-
Lot 2	60-/0+	60-/0+	60-/0+	60-/0+	28-/32+	60+/0-	60+/0-	60+/0-	60+/0-
Lot 3	60-/0+	60-/0+	60-/0+	59-/1+	29-/31+	60+/0-	60+/0-	60+/0-	60+/0-

c. Stability

The devices are stable at 2-30°C for 24 months based on the accelerated stability study at 45°C. The real time stability study is ongoing.

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

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concentration of 100µg/mL or specified concentrations are summarized in the following tables.

Acetaminophen	Doxepin (50 µg/mL)	Nortriptyline (25 µg/mL)
Acetone (1000 mg/dL)	Ecgonine methyl ester	Noscapine
Acetophenetidin	Ephedrine	O-Hydroxyhippuric acid
Acetylsalicylic acid	Erythromycin	Octopamine
Albumin (100 mg/dL)	Ethanol (1%)	Oxalic acid (100 mg/dL)
Albuterol	Fenoprofen	Oxazepam
Aminopyrine	Fluphenazine	Oxolinic acid
Amitriptyline (35 µg/mL)	Furosemide	Oxymetazoline
Amobarbital	Galactose (10 mg/dL)	Papaverine
Amoxicillin	Gamma Globulin (500 mg/dL)	Penicillin G
Ampicillin	Gentisic acid	Perphenazine
Apomorphine	Glucose (3000 mg/dL)	Phencyclidine
Ascorbic acid	Hemoglobin	Phenelzine
Aspartame	Hydralazine	Phenobarbital
Atropine	Hydrochlorothiazide	Prednisone
Benzilic acid	Hydrocortisone	Propoxyphene (50 µg/mL)
Benzoic acid	Hydroxytyramine	Propranolol
Benzoylecgonine	Ibuprofen	Pseudoephedrine
Bilirubin	Imipramine (30 µg/mL)	Quinine
Boric Acid (1%)	Isoproterenol	Ranitidine
Bupropion (50 µg/mL)	Isoxsuprine	Riboflavin (7.5 mg/dL)
Caffeine	Ketamine	Salicylic acid
Carbamazepine	Ketoprofen	Secobarbital
Chloral hydrate	Labetalol	Serotonin (5-Hydroxytyramine)
Chloramphenicol	Lidocaine (50 µg/mL)	Sulfamethazine
Chlorothiazide	Loperamide	Sulindac
Chlorpromazine	Maprotiline (50 µg/mL)	Tetrahydrocortisone 3-(β-Dglucuronide)
Cholesterol	Meperidine	Tetrahydrocortisone 3-acetate
Clomipramine (50 µg/mL)	Meprobamate	Tetrahydrozoline
Clonidine	Methapyrilene (10 µg/mL)	Thiamine
Cortisone	Methaqualone (50 µg/mL)	Thioridazine
Cotinine	Methoxyphenamine	Triamterene
Creatinine	Metronidazole (300 µg/mL)	Trifluoperazine
Cyclobenzaprine (10 µg/mL)	N-Acetylprocainamide	Trimethoprim
Deoxycorticosterone	NaCl (4000 mg/dL)	Tyramine
Desipramine (50 µg/mL)	Nalidixic acid	Urea (2000 mg/dL)
Dextromethorphan	Naloxone	Uric acid
Diclofenac	Naltrexone	Valproic acid (250 µg/mL)
Diflunisal	Naproxen	Venlafaxine
Digoxin	Niacinamide	Verapamil
Diphenhydramine	Nicotine (10 µg/mL)	Zomepirac
DL-Tryptophan	Nifedipine	β-Estradiol
DL-Tyrosine	Norethindrone

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.

Drug	Concentration(ng/mL)	% Cross-Reactivity
Acetyl fentanyl	1.2	83.33
Acrylfentanyl	1.2	83.33

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Drug	Concentration(ng/mL)	% Cross-Reactivity
ω-1-Hydroxyfentanyl	20000	0.01
Isobutyryl fentanyl	1.5	66.67
Ocfentanil	1.5	66.67
Butyryl fentanyl	1.6	62.50
Furanyl fentanyl	1.75	57.14
Valeryl fentanyl	2.5	40.00
(±) β-hydroxythiofentanyl	2.8	35.71
4-Fluoro-isobutyrylfentanyl	3	33.33
Para-fluorobutyrylfentanyl	3	33.33
Para-fluoro fentanyl	3	33.33
(±)-3-cis-methylfentanyl	5	20.00
Carfentanil	500	0.20
Sufentanil	625	0.16
Alfentanil	100000	0.00
Despropionyl fentanyl(4-ANPP)	50000	0.00

The following other Metabolites and opioids compounds were tested at a concentration of 100 µg/mL. Negative results were obtained for all these compounds. There is no cross-reactivity for these compounds using the BioSieve™ Fentanyl FIA Test Kit.

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

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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 target fentanyl at 50% below and 50% above Cut-Off levels. by three different operators per lot of device, with a total of three lots. Results were all positive for samples at and above +50% Cut-Off and all negative for samples at and below -50% Cut-Off.
1. Comparison Studies
  Method comparison studies for the BioSieve™ Fentanyl FIA Test Kit. were performed at three different testing sites. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples. The samples were blind labeled and compared to LC-MS/MS results. The results are presented in the tables below.

		Negative	LowNegative byLC-MS/MS(less than-50%)	Near CutoffNegative byLC-MS/MS(Between-50% andcutoff)	Near CutoffPositive byLC-MS/MS(Between thecutoff and+50%)	High Positiveby LC-MS/MS(greater than+50%)
Site1	Positive	0	0	4	19	20
	Negative	10	18	8	1	0
Site2	Positive	0	0	3	18	20
	Negative	10	18	9	2	0
Site3	Positive	0	0	2	19	20
	Negative	10	18	10	1	0

Discordant Results

Operator	Sample Number	LC-MS/MSResult	BioSieveResults
Site 1	VCFC021	0.849	Positive
Site 1	VCFC075	0.875	Positive
Site 1	VCFC059	0.985	Positive
Site 1	VCFC048	0.946	Positive
Site 1	VCFC057	1.063	Negative
Site 2	VCFC030	0.892	Positive
Site 2	VCFC010	0.934	Positive
Site 2	VCFC059	0.985	Positive
Site 2	VCFC057	1.063	Negative
Site 2	VCFC036	1.104	Negative
Site 3	VCFC010	0.934	Positive
Site 3	VCFC048	0.946	Positive
Site 3	VCFC070	1.129	Negative

1. Clinical Studies

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Not applicable.

11. Conclusion

Based on the test principle and acceptable performance characteristics including precision, cut-off, interference, specificity, and method comparison 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).