Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device is a simple lateral flow immunoassay, which relies on chemical reactions and visual interpretation, not AI/ML. The text explicitly states "Not Found" for mentions of AI, DNN, or ML.

Therapeutic

No
The device is an in vitro diagnostic test for detecting fentanyl in urine, not for treating any condition.

Diagnostic

Yes

The text explicitly states that the device is for "in vitro diagnostic use only" and describes its purpose as the "qualitative detection of fentanyl in human urine," which are characteristics of a diagnostic device.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the device consists of a Test Cassette, a Dropper, and a package insert, which are physical components, not software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

The intended use statement explicitly states: "The test is intended for over-the-counter use and for in vitro diagnostic use only."

Furthermore, the device performs a test on a human sample (urine) to detect a substance (fentanyl) for diagnostic purposes (preliminary detection of fentanyl use). This aligns with the definition of an in vitro diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

SAFECARE Fentanyl Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method. The test is intended for over-the-counter use and for in vitro diagnostic use only.

SAFECARE FYL Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method.

Product codes (comma separated list FDA assigned to the subject device)

NGL

Device Description

The SAFECARE Fentanyl Urine Tests are immunoassays intended for the qualitative detection of fentanyl in human urine. Each SAFECARE Fentanyl Urine Test device consists of a Test Cassette, a Dropper and a package insert. Each Test Cassette is sealed with sachets of desiccant in an aluminum pouch.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Human urine

Indicated Patient Age Range

Not Found

Intended User / Care Setting

over-the-counter use and for in vitro diagnostic use only.

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Precision Test:

Sample Size: For each concentration, 50 tests were performed (two tests per day for 25 days per device lot). Total of 3 lots tested, meaning 150 tests per concentration across all lots.
Data Source: Samples were prepared by spiking fentanyl in negative samples. Fentanyl concentrations were confirmed by LC/MS. Sample aliquots were blindly labeled.
Annotation Protocol: Not explicitly stated, but results are observed as positive (+) or negative (-) relative to the cutoff.

Interference Test:

Sample Size: Not explicitly stated, but "three batches of each device" were used.
Data Source: Potential interfering substances were added to drug-free urine and target drug fentanyl urine (at 50% below and 50% above Cut-Off levels).
Annotation Protocol: Not explicitly stated, but interference was determined by observation of positive or negative results.

Specificity Test:

Sample Size: Not explicitly stated, but "three batches of device" were used.
Data Source: Drug metabolites and other components likely to interfere in urine samples.
Annotation Protocol: The lowest concentration that caused a positive result for each compound was listed. Cross-reactivity was calculated.

Effect of Urine Specific Gravity and Urine pH:

Sample Size: Not explicitly stated, but "three lots of device" were used.
Data Source: Urine samples with specific gravity from 1.000 to 1.035 or pH from 4 to 9, spiked with target fentanyl at 50% below and 50% above Cut-Off levels.
Annotation Protocol: Results were observed as positive or negative.

Method Comparison Studies for the SAFECARE Fentanyl Urine Test:

Sample Size: 80 human urine samples (40 negative and 40 positive).
Data Source: Unaltered clinical samples, blindly labeled.
Annotation Protocol: Samples were compared to LC/MS results. Results are presented as positive/negative calls by operators vs. LC/MS categories (Negative, Low Negative, Near Cutoff Negative, Near Cutoff Positive, High Positive). Discordant results were individually listed with LC/MS result and Rapid Test Result.

Lay-user study:

Sample Size: 140 lay persons.
Data Source: Urine samples prepared at concentrations of -100%, -75%, -50%, -25%, +25%, +50%, +75% of the cut-off by spiking fentanyl into drug-free pooled urine specimens. Concentrations were confirmed by LC/MS. Each sample was aliquoted, blind-labeled, and randomized.
Annotation Protocol: Each participant was given one blind-labeled sample and a device. Results (number of positive, number of negative) were recorded, and the percentage of correct results was calculated. A survey on ease of understanding the package insert instructions was also administered.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Analytical Performance - Precision:

Study Type: Precision study.
Sample Size: 50 tests per concentration per lot, across 3 lots.
Key Results:
- -100%, -75%, -50% cutoff: All 50 tests per lot were negative (50-/0+).
- -25% cutoff: Lot 1: 49-/1+; Lot 2: 49-/1+; Lot 3: 48-/2+.
- Cutoff: Lot 1: 26+/24-; Lot 2: 25+/25-; Lot 3: 27+/23-.
- +25%, +50%, +75%, +100% cutoff: All 50 tests per lot were positive (50+/0-).

Analytical Performance - Interference:

Study Type: Interference study.
Key Results: A comprehensive list of substances (e.g., Acetaminophen, Ethanol, Ibuprofen, Nicotine) showed no interference at 100µg/mL or specified concentrations.

Analytical Performance - Specificity:

Study Type: Specificity study.
Key Results: Cross-reactivity was determined for various fentanyl derivatives and other opioids. Examples include: Acetyl fentanyl (1 ng/mL, 100% cross-reactivity), Ocfentanil (2.5 ng/mL, 40% cross-reactivity), Carfentanil (2 ng/mL, 50% cross-reactivity), Sufentanil (7.5 ng/mL, 13.3% cross-reactivity), Remifentanil (100 ng/mL, 1% cross-reactivity). Many other opioids (e.g., 6-Acetyl morphine, Amphetamine, Morphine) showed no cross-reactivity at 100ug/mL.

Analytical Performance - Effect of Urine Specific Gravity and Urine pH:

Study Type: Urine specific gravity and pH effect study.
Key Results: Results were all positive for samples at and above +50% Cut-Off and all negative for samples at and below -50% Cut-Off across the tested ranges of specific gravity (1.000 to 1.035) and pH (4 to 9).

Comparison Studies:

Study Type: Method comparison studies with clinical samples.
Sample Size: 80 clinical samples (40 negative, 40 positive) for each of three operators (total 240 tests).
Key Results:
- Operator 1:
  - Positive: 0 (Negative), 0 (Low Negative), 1 (Near Cutoff Negative), 18 (Near Cutoff Positive), 20 (High Positive)
  - Negative: 10 (Negative), 10 (Low Negative), 19 (Near Cutoff Negative), 2 (Near Cutoff Positive), 0 (High Positive)
- Operator 2:
  - Positive: 0 (Negative), 0 (Low Negative), 2 (Near Cutoff Negative), 19 (Near Cutoff Positive), 20 (High Positive)
  - Negative: 10 (Negative), 10 (Low Negative), 18 (Near Cutoff Negative), 1 (Near Cutoff Positive), 0 (High Positive)
- Operator 3:
  - Positive: 0 (Negative), 0 (Low Negative), 2 (Near Cutoff Negative), 19 (Near Cutoff Positive), 20 (High Positive)
  - Negative: 10 (Negative), 10 (Low Negative), 18 (Near Cutoff Negative), 1 (Near Cutoff Positive), 0 (High Positive)
- Discordant Results: A few samples showed discordance, mainly near the cutoff concentration (e.g., sample FM157, LC/MS 0.985 ng/mL, Rapid Test Positive for Operator 1 and 3; sample FM152, LC/MS 1.055 ng/mL, Rapid Test Negative for all three operators).

Lay-user study:

Study Type: Lay-user study.
Sample Size: 140 lay persons.
Key Results:
- -100%, -75%, -50% Cutoff: 100% correct negative results (20/20 samples).
- -25% Cutoff: 95% correct negative results (19/20 samples, 1 positive result).
- +25%, +50%, +75% Cutoff: 100% correct positive results (20/20 samples).
- All lay users indicated that the device instructions can be easily followed. The package insert had a Flesch-Kincaid reading grade level of less than 7.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Not explicitly stated as calculated metrics like sensitivity, specificity, PPV, NPV. Precision data and correct result percentages from the lay-user study are provided directly.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

AllTest Fentanyl Urine Test Cassette (K233417)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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).

Summary

0

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left, there is a symbol representing the Department of Health & Human Services USA. To the right of the symbol, there is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of the word "ADMINISTRATION."

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

Re: K240654

Trade/Device Name: SAFECARE Fentanyl Urine Test Cassette; SAFECARE FYL Urine Test Cassette Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: NGL Dated: March 2, 2024 Received: March 7, 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.

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

1

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 medical devices and radiation-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.
Kotarek -S

Digitally signed by Joseph A. Kotarek -S Date: 2024.04.10 16:06:14 -04'00'

Joseph Kotarek Branch Chief for Toxicology 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

2

Indications for Use

510(k) Number (if known) K240654

Device Name SAFECARE Fentanyl Urine Test Cassette SAFECARE FYL Urine Test Cassette

Indications for Use (Describe)

SAFECARE Fentanyl Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method. The test is intended for over-the-counter use and for in vitro diagnostic use only.

SAFECARE FYL Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL.

The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method.

Type of Use (Select one or both, as applicable)
-------------------------------------------------	--

Prescription Use (Part 21 CFR 801 Subpart D)

|X | Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) SUMMARY K240654

1. Date:	April 8, 2024
2. Submitter:	Safecare Biotech (Hangzhou) Co. Ltd.
18 Haishu Road, Yuhang District
Hangzhou, China
3. Contact person:	Joe Shia
LSI International Inc.
504E Diamond Ave., Suite H
Gaithersburg, MD 20877
Telephone: 240-505-7880
Email: shiajl@yahoo.com
4. Device Names:	SAFECARE Fentanyl Urine Test Cassette
SAFECARE FYL Urine Test Cassette

Classification:	Class 2
Product Code	Classification	Regulation Section	Panel
NGL	II	21 CFR § 862.3650
Opiate Test System	Toxicology (91)

1. Predicate Devices:
  AllTest Fentanyl Urine Test Cassette (K233417)
1. Indications for Use
  SAFECARE Fentanyl Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL.

The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method.

The test is intended for over-the-counter use and for in vitro diagnostic use only.

SAFECARE FYL Urine Test Cassette is a competitive binding, lateral flow immunochromatographic assay for qualitative detection of fentanyl in human urine at the cutoff concentrations of 1 ng/mL.

The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS or LC/MS is the preferred confirmatory method.

1. Device Description
  The SAFECARE Fentanyl Urine Tests are immunoassays intended for the qualitative detection of fentanyl in human urine. Each SAFECARE Fentanyl Urine Test device consists of a Test Cassette, a Dropper and a package insert. Each Test Cassette is sealed with sachets of desiccant in an aluminum pouch.
1. Substantial Equivalence Information

4

A summary comparison of features of the SAFECARE Fentanyl Test and the predicate devices is provided in following table.

Item	Device	Predicate – K233417
Indication(s)
for Use	For the qualitative determination of
fentanyl in human urine.	Same
Calibrator and Cut-Off
Values	Fentanyl (FTY)
1 ng/ml	Same
Methodology	Competitive binding, lateral flow
immunochromatographic assays based
on the principle of antigen antibody
immunochemistry.	Same
Type of Test	Qualitative	Same
Specimen Type	Human Urine	Same
Intended Use	For OTC use	Same
Configurations	Cassette	Same
Storage	4-30°C	Same

Table 1: Features Comparison of SAFECARE Fentanyl Urine Test and the Predicate Device

9. Test Principle

The SAFECARE Fentanyl Urine Tests are immunoassays based on the principle of competitive binding.

During testing, a urine specimen migrates upward by capillary action. Fentanyl, if present in the urine specimen below 1 ng/mL, will not saturate the binding sites of antibody-coated particles in the test device. The antibody-coated particles will then be captured by immobilized fentanyl 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 fentanyl level exceeds 1 ng/mL because it will saturate all the binding sites of anti-fentanyl 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.

10. Performance Characteristics

1. Analytical Performance
- Precision a.

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. 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 two tests per day for 25 days per device lot in a randomized order.

5

| Lot
Number | -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 | 50-/0+ | 50-/0+ | 50-/0+ | 49-/1+ | 26+/24- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 2 | 50-/0+ | 50-/0+ | 50-/0+ | 49-/1+ | 25+/25- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot 3 | 50-/0+ | 50-/0+ | 50-/0+ | 48-/2+ | 27+/23- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

c. Stability

The devices are stable at 36-86F for 24 months based on the accelerate 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	Doxepin	Nortriptyline
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	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
Benzoic acid	Hydroxytyramine	Propranolol
Benzoylecgonine	Ibuprofen	Pseudoephedrine
Bilirubin	Imipramine	Quinine
Boric acid (1% w/v)	Isoproterenol	Ranitidine
Bupropion	Isoxsuprine (10ug/mL)	Riboflavin (10 mg/dL)
Caffeine	Ketamine	Salicylic acid
Carbamazepine	Ketoprofen	Secobarbital
Chloral hydrate	Labetalol	Serotonin (5-hydroxytyramine)
Chloramphenicol	Lidocaine	Sulfamethazine
Chlorothiazide	Loperamide	Sulindac
Chlorpromazine	Maprotiline	Tetrahydrocortisone 3-(β-Dglucuronide)
Cholesterol	Meperidine	Tetrahydrocortisone 3-acetate
Clomipramine	Meprobamate	Tetrahydrozoline
Clonidine	Methapyrilene	Thiamine
Cortisone	Methaqualone	Thioridazine
Cotinine	Methoxyphenamine	Triamterene
Creatinine	Metronidazole (300 ug/mL)	Trifluoperazine

6

Cyclobenzaprine	N-Acetylprocainamide	Trimethoprim
Deoxycorticosterone	NaCl (4000 mg/dL)	Tyramine
Desipramine	Nalidixic acid	Urea (2000 mg/dL)
Dextromethorphan	Naloxone	Uric acid
Diclofenac	Naltrexone	Valproic acid (250 ug/mL)
Diflunisal	Naproxen	Venlafaxine
Digoxin	Niacinamide	Verapamil
Diphenhydramine	Nicotine	Zomepirac
DL-Tryptophan	Nifedipine	B-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.

| Fentanyl (Cutoff=1ng/mL) concentration required
to obtain a positive
result (ng/mL) | % Cross-Reactivity |---------------------------- | Acetyl fentanyl | Acrylfentanyl | ω-1-Hydroxyfentanyl | Isobutyryl fentanyl | Ocfentanil | Butyryl fentanyl | Furanyl fentanyl | Valeryl fentanyl | (±) β-hydroxythiofentanyl | 4-Fluoro-isobutyrylfentanyl | 50 | Para-fluorobutyryl fentanyl | 20 | Para-fluoro fentanyl | (±)-3-cis-methyl fentanyl | Carfentanil | Sufentanil | Alfentanil | Despropionyl fentanyl (4-ANPP) | 2500 | Remifentanil | Norfentanyl | Acetyl norfentanyl | Norcarfentanil | Trazodone | Minimum
|
----|-----------------------------------------------------------------------------|--------------------|
| 1 | 100 |
| 10 | 10 |
| 20000 | 0.005 |
| 10 | 10 |
| 2.5 | 40 |
| 10 | 10 |
| 10 | 10 |
| 10 | 10 |
| 6 | 16.7 |
| 2 |
| 5 |
| 3 | 33.3 |
| 50 | 2 |
| 2 | 50 |
| 7.5 | 13.3 |
| 500 | 0.2 |
| 0.04 |
| 100 | 1 |
| 2000 | 0.05 |
| >100000 | / |
| >100000 | / |
| >100000 | / |

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 SAFECARE Fentanyl Test.

6-Acetyl morphine	Naltrexone
Amphetamine	Norbuprenorphine
Buprenorphine	Norcodeine

7

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

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. These samples were tested using three lots of device. Results were all positive for samples at and above +50% Cut-Off and all negative for samples at and below -50% Cut-Off.

2. Comparison Studies

Method comparison studies for the SAFECARE Fentanyl Urine 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. The results are presented in the tables below.

| | | Negative | Low
Negative by
LC/MS
(less than
-50%) | Near Cutoff
Negative by
LC/MS
(Between
-50% and
cutoff) | Near Cutoff
Positive by
LC/MS
(Between the
cutoff and
+50%) | High Positive
by LC/MS
(greater than
+50%) |
|----------|----------|----------|----------------------------------------------------|------------------------------------------------------------------------|----------------------------------------------------------------------------|-----------------------------------------------------|
| Operator | Positive | 0 | 0 | 1 | 18 | 20 |
| 1 | Negative | 10 | 10 | 19 | 2 | 0 |
| Operator | Positive | 0 | 0 | 2 | 19 | 20 |
| 2 | Negative | 10 | 10 | 18 | 1 | 0 |
| Operator | Positive | 0 | 0 | 2 | 19 | 20 |
| 3 | Negative | 10 | 10 | 18 | 1 | 0 |

Discordant Results
Operator	Sample ID	LC/MS Result (ng/mL)	Rapid Test Result
Operator 1	FM157	0.985	Positive
Operator 1	FM152	1.055	Negative

8

Operator 1	FM212	1.097	Negative
Operator 2	FM083	0.954	Positive
Operator 2	FM024	0.993	Positive
Operator 2	FM152	1.055	Negative
Operator 3	FM179	0.980	Positive
Operator 3	FM157	0.985	Positive
Operator 3	FM152	1.055	Negative

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 20 to >50 years. Urine samples were prepared at the following concentrations; -100%, +/-50%, +/-50%, +/-25% of the cut-off by spiking fentanyl 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
of
samples | Fentanyl Concentration
by LC/MS
(ng/mL) | Lay person results | | The
percentage of |
|--------------|-------------------------|-----------------------------------------------|--------------------|--------------------|------------------------|
| | | | No. of
Positive | No. of
Negative | correct results
(%) |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100 |
| -75% Cutoff | 20 | 0.24 | 0 | 20 | 100 |
| -50% Cutoff | 20 | 0.46 | 0 | 20 | 100 |
| -25% Cutoff | 20 | 0.71 | 1 | 19 | 95 |
| +25% Cutoff | 20 | 1.31 | 20 | 0 | 100 |
| +50% Cutoff | 20 | 1.58 | 20 | 0 | 100 |
| +75% Cutoff | 20 | 1.77 | 20 | 0 | 100 |

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.

4. Clinical Studies

Not applicable.

11. 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.