(20 days)
The LZI Fentanyl III Enzyme Immunoassay is intended for the qualitative determination of fentanyl in human urine at the cutoff value of 1 ng/mL when calibrated against fentanyl. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive.
The LZI Fentanyl III Enzyme Immunoassay is a homogeneous enzyme immunoassay with ready-to-use liquid reagents. The assay is based on competition between the drug in the sample and the drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. The drug-labeled G6PDH conjugate is traceable to a commercially available fentanyl standard and referred to as fentanyl-labeled G6PDH conjugate. Enzyme activity decreases upon binding to the antibody, and the drug concentration in the sample is measured in terms of enzyme activity. In the absence of a drug in the sample, fentanyl-labeled G6PDH conjugate is bound to the antibody, and the enzyme activity is inhibited. On the other hand, when the free drug is present in the sample, the antibody would bind to the free drug; the unbound fentanyl-labeled G6PDH then exhibits its maximal enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at 340 nm.
The LZI Fentanyl III Enzyme Immunoassay is a kit comprised of two reagents, R1 and R2, which are bottled separately but sold together within the kit.
The R1 solution contains mouse monoclonal anti-fentanyl antibody, glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide (NAD), stabilizers, and sodium azide (0.09%) as a preservative. The R2 solution contains glucose-6-phosphate dehydrogenase (G6PDH) labeled with fentanyl in buffer with sodium azide (0.09%) as a preservative.
The provided FDA 510(k) Clearance Letter for the LZI Fentanyl III Enzyme Immunoassay details the device's acceptance criteria (in terms of performance characteristics) and the study results that demonstrate the device meets these criteria.
Here's a breakdown of the requested information:
1. Acceptance Criteria and Reported Device Performance
The acceptance criteria are implicitly defined by the satisfactory performance demonstrated in the various studies, aiming to prove substantial equivalence to the predicate device. The performance characteristics sections show the device meets the expected qualitative and quantitative accuracy, precision, and specificity.
| Acceptance Criteria Category | Specific Criteria (Implicitly Derived from Study Design/Outcomes) | Reported Device Performance |
|---|---|---|
| Intended Use Equivalence | Device is substantially equivalent to the predicate (LZI Fentanyl II Enzyme Immunoassay) in terms of intended use, method principle, device components, and clinical performance. | The submission states, "The LZI Fentanyl III Enzyme Immunoassay is substantially equivalent to the LZI Fentanyl II Enzyme Immunoassay (K201938) manufactured by LZI in terms of intended use, method principle, device components, and clinical performance." The changes are clearly identified (target analyte from norfentanyl to fentanyl, updated cutoff, and assay application parameters), and subsequent studies address the impact of these changes. |
| Qualitative Accuracy | High concordance with LC/MS confirmation for true positives and true negatives, especially at and around the cutoff concentration. | Method Comparison - Clinical Samples (1 ng/mL Cutoff):- Negative by LC/MS analysis: 35 negatives, 0 positives by EIA.- < 50% of cutoff by LC/MS: 0 negatives, 20 positives by EIA (discrepant, attributed to norfentanyl cross-reactivity).- Near Cutoff Negative (50% below to cutoff) by LC/MS: 0 negatives, 12 positives by EIA (discrepant, attributed to norfentanyl cross-reactivity).- Near Cutoff Positive (cutoff to 50% above) by LC/MS: 0 negatives, 22 positives by EIA.- High Positive (> 50% above cutoff) by LC/MS: 0 negatives, 61 positives by EIA.Overall, 35/35 true negatives, 83/83 true positives (excluding near-cutoff positives and positives above cutoff). The remaining 32 discrepant samples (20 + 12) were positive by EIA but negative by LC/MS for fentanyl, indicating cross-reactivity with norfentanyl as a contributing factor. |
| Precision | Consistent and reliable qualitative results (positive/negative) across multiple runs and days, especially around the cutoff concentration. | Precision: 1 ng/mL Cutoff (N=88 total replicates over 22 days):- 0 ng/mL, 0.25 ng/mL, 0.5 ng/mL, 0.75 ng/mL: 88/88 Negative (100% agreement).- 1 ng/mL (Cutoff): 4 Neg / 84 Pos (95.5% Positive, 4.5% Negative - demonstrating near-cutoff variability, which is expected for qualitative assays at the cutoff).- 1.25 ng/mL, 1.5 ng/mL, 1.75 ng/mL, 2 ng/mL: 88/88 Positive (100% agreement). |
| Cross-Reactivity (Specificity) | Minimal or no false positives from structurally unrelated compounds. Acceptable cross-reactivity with fentanyl metabolites and structurally related compounds as defined by the assay's intended specificity. | Fentanyl and Metabolites: Fentanyl (100%), Norfentanyl (40%).Structurally Related Compounds: Varying levels of cross-reactivity (e.g., Acetyl fentanyl 25%, Acryl fentanyl 100%, Thiofentanyl 200%). Many substances showed "ND" (Not Detected) at high concentrations (e.g., 100,000 ng/mL).Structurally Unrelated Pharmacological Compounds: No interference (all Neg/Neg/Pos results for 0 ng/mL Fentanyl, -50% Cutoff, +50% Cutoff respectively) at 100,000 ng/mL for almost all listed compounds, except Dextromethorphan, which showed interference at 20,000 ng/mL (Neg/Pos/Pos) but was acceptable at 15,000 ng/mL (Neg/Neg/Pos). This demonstrates good specificity against common drugs. |
| Interference | Test performance (positive/negative results) should not be significantly affected by common endogenous substances or preservatives found in urine, or by varying specific gravity and pH within physiological ranges. | Endogenous and Preservative Compound Interference: No interference observed for most compounds (e.g., Acetone, Ascorbic acid, Bilirubin, Glucose, Hemoglobin) at high concentrations (e.g., 100,000 mg/dL), showing Neg/Neg/Pos results for 0 ng/mL, -50% Cutoff, +50% Cutoff respectively. Boric acid showed interference at 1,000 mg/dL (Neg/Neg/Neg), indicating a potential issue if present at high concentrations. This is a known interference for certain urine assays.Specific Gravity Interference: No interference observed across the range of 1.000 to 1.030 (all Neg/Neg/Pos).pH Interference: No major interference observed between pH 3 to pH 11 (all Neg/Neg/Pos). |
2. Sample Size and Data Provenance
- Test Set Sample Size:
- Precision Study: 88 replicates for each concentration level (0 ng/mL to 2 ng/mL fentanyl).
- Method Comparison (Clinical Samples): 150 unaltered clinical samples.
- Cross-Reactivity: Duplicates for each compound and concentration tested.
- Interference (Endogenous/Preservative): Duplicates for each compound and concentration tested.
- Specific Gravity/pH Interference: Duplicates for each specific gravity/pH level and fentanyl concentration.
- Data Provenance:
- The clinical samples for the Method Comparison study were obtained by LZI and "through collaboration with various clinical laboratories across the United States and Canada." This indicates a prospective and/or retrospective collection of real-world clinical samples from diverse geographical regions. The nature (retrospective/prospective) of the collection for these specific 150 samples isn't explicitly stated but "clinical samples" usually implies samples collected from patients.
3. Number of Experts and Qualifications for Ground Truth
- The document does not specify the number of experts used to establish ground truth.
- Qualifications of Experts: Not explicitly stated. However, for LC/MS confirmation, it implicitly relies on the expertise of the laboratory personnel performing and interpreting the LC/MS (Liquid Chromatography-Mass Spectrometry) analysis, which is a gold standard for chemical identification and quantification. These would typically be trained analytical chemists or lab technicians with experience in mass spectrometry.
4. Adjudication Method for the Test Set
- The document does not describe an adjudication method for the test set in the typical sense of multiple human readers or reviewers resolving discrepancies.
- For the Method Comparison study, the "ground truth" was established independently by LC/MS. Discrepancies between the EIA result and the LC/MS result were reported and attributed to norfentanyl cross-reactivity. This is a comparison against a reference method, not a subjective adjudication by experts.
5. Multi Reader Multi Case (MRMC) Comparative Effectiveness Study
- No, an MRMC comparative effectiveness study was not done. This type of study is more common for diagnostic imaging AI algorithms where human interpretation is a primary component and AI aims to augment or replace it. This document describes an in vitro diagnostic (IVD) assay, an enzyme immunoassay, which is an automated lab test. Its performance is evaluated against a reference standard (LC/MS), not against human reader performance.
6. Standalone Performance
- Yes, standalone performance was done. The entire submission details the performance of the LZI Fentanyl III Enzyme Immunoassay as a standalone device. The qualitative accuracy, precision, cross-reactivity, and interference studies all evaluate the algorithm/device's performance independently in generating a preliminary analytical result from a urine sample. It does not involve human-in-the-loop performance; rather, it provides a result that a clinician then interprets in conjunction with other clinical data.
7. Type of Ground Truth Used
- The primary type of ground truth used for the quantitative confirmation of fentanyl concentration in the clinical samples (Method Comparison study) was LC/MS (Liquid Chromatography-Mass Spectrometry) analysis. This is considered a highly specific and sensitive reference method for drug quantification.
8. Sample Size for the Training Set
- The document does not explicitly state the sample size for the training set. The described studies are verification and validation activities for a modified assay (Special 510(k) for an existing predicate device, LZI Fentanyl II). For IVD assays, "training" often refers to the internal development and optimization of the assay performed by the manufacturer, which precedes the formal V&V studies presented for regulatory submission. The reported studies are the test set performance.
9. How the Ground Truth for the Training Set Was Established
- Since the training set size is not explicitly stated, the method for establishing ground truth for it is also not explicitly described. However, it is highly probable that internal development and optimization of the assay would have utilized similar rigorous analytical methods, likely LC/MS or other established analytical techniques, to calibrate and refine the assay's performance against known concentrations of fentanyl and its metabolites. This would be part of the manufacturer's design control and development process.
FDA 510(k) Clearance Letter - LZI Fentanyl III Enzyme Immunoassay
Page 1
U.S. Food & Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993
www.fda.gov
Doc ID # 04017.07.05
June 18, 2025
Lin-Zhi International, Inc.
Chris Wang
Senior Manager, DAU Assay Development
2945 Oakmead Village Court
Santa Clara, California 95051
Re: K251634
Trade/Device Name: LZI Fentanyl III Enzyme Immunoassay
Regulation Number: 21 CFR 862.3650
Regulation Name: Opiate Test System
Regulatory Class: Class II
Product Code: DJG
Dated: May 28, 2025
Received: May 29, 2025
Dear Chris Wang:
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.
U.S. Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993
www.fda.gov
Page 2
K251634 - Chris Wang Page 2
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 (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-safety-reporting-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-devices/device-advice-comprehensive-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-devices/medical-device-safety/medical-device-reporting-mdr-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/medical-devices/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-devices/device-advice-comprehensive-regulatory-
Page 3
K251634 - Chris Wang Page 3
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,
JOSEPH A. KOTAREK -S
Digitally signed by JOSEPH A. KOTAREK -S
Date: 2025.06.18 14:21:24 -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
Page 4
FORM FDA 3881 (8/23) Page 1 of 1
DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
Indications for Use
Form Approved: OMB No. 0910-0120
Expiration Date: 07/31/2026
See PRA Statement below.
510(k) Number (if known): K251634
Device Name: LZI Fentanyl III Enzyme Immunoassay
Indications for Use (Describe):
The LZI Fentanyl III Enzyme Immunoassay is intended for the qualitative determination of fentanyl in human urine at the cutoff value of 1 ng/mL when calibrated against fentanyl. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse 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)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services
Food and Drug Administration
Office of Chief Information Officer
Paperwork Reduction Act (PRA) Staff
PRAStaff@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
Page 5
Lin-Zhi International, Inc.
510(k) Summary
510(k) Number: K251634
Prepared On: June 17, 2025
Submitter Name and Contact Person:
Chris Wang, M.S.
Senior Manager, DAU Assay Development
Phone: (408) 970-8811
Fax: (408) 970-9030
E-mail: cjwang@lin-zhi.com
Company Address:
Lin-Zhi International, Inc.
2945 Oakmead Village Court
Santa Clara, CA 95051
Page 6
Lin-Zhi International, Inc.
Introduction
This submission is provided in accordance with 21 CFR 807.92 as a Special 510(k) for the LZI Fentanyl III Enzyme Immunoassay. The purpose of this application is to support modifications made to LZI's legally marketed predicate device, the LZI Fentanyl II Enzyme Immunoassay (K201938). These modifications include a change in target analyte from norfentanyl to fentanyl, an updated cutoff concentration, and changes to assay application parameters.
Verification and validation activities were conducted using well-established methods consistent with those performed for the predicate device. These included method comparison with LC/MS, precision studies, cross-reactivity evaluation, and interference testing. Collectively, the data confirm that the modified device supports its intended use, safety, and effectiveness, and performs substantially equivalent to the predicate.
This submission includes a summary of design control activities and performance characteristics, which together provide a complete basis for a determination of substantial equivalence.
Device Name and Classification
Classification Name: Enzyme Immunoassay, Opiates
Regulation Number: 21 CFR 862.3650
Product Code: Class II, DJG (91 Toxicology)
Common Name: Homogeneous Fentanyl Enzyme Immunoassay
Proprietary Name: LZI Fentanyl III Enzyme Immunoassay
Submission Type: Special 510(k)
510(k) Number: K251634
Legally Marketed Predicate Device
The subject device is compared to the predicate:
Predicate Device: LZI Fentanyl II Enzyme Immunoassay
510(k) Number: K201938
The LZI Fentanyl III Enzyme Immunoassay is substantially equivalent to the LZI Fentanyl II Enzyme Immunoassay (K201938) manufactured by LZI in terms of intended use, method principle, device components, and clinical performance.
Page 7
Lin-Zhi International, Inc.
Device Description
The LZI Fentanyl III Enzyme Immunoassay is a homogeneous enzyme immunoassay with ready-to-use liquid reagents. The assay is based on competition between the drug in the sample and the drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. The drug-labeled G6PDH conjugate is traceable to a commercially available fentanyl standard and referred to as fentanyl-labeled G6PDH conjugate. Enzyme activity decreases upon binding to the antibody, and the drug concentration in the sample is measured in terms of enzyme activity. In the absence of a drug in the sample, fentanyl-labeled G6PDH conjugate is bound to the antibody, and the enzyme activity is inhibited. On the other hand, when the free drug is present in the sample, the antibody would bind to the free drug; the unbound fentanyl-labeled G6PDH then exhibits its maximal enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at 340 nm.
The LZI Fentanyl III Enzyme Immunoassay is a kit comprised of two reagents, R1 and R2, which are bottled separately but sold together within the kit.
The R1 solution contains mouse monoclonal anti-fentanyl antibody, glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide (NAD), stabilizers, and sodium azide (0.09%) as a preservative. The R2 solution contains glucose-6-phosphate dehydrogenase (G6PDH) labeled with fentanyl in buffer with sodium azide (0.09%) as a preservative.
Intended Use
The LZI Fentanyl III Enzyme Immunoassay is intended for the qualitative determination of fentanyl in human urine at the cutoff value of 1 ng/mL when calibrated against fentanyl. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive.
Page 8
Lin-Zhi International, Inc.
Substantial Equivalence Comparison to Predicate Device
The LZI Fentanyl III Enzyme Immunoassay is substantially equivalent to the LZI Fentanyl II Enzyme Immunoassay which was cleared by the FDA under the premarket notification K201938 for its stated intended use.
The following table compares the LZI Fentanyl III Enzyme Immunoassay with the predicate device.
| Device Characteristics | Subject DeviceLZI Fentanyl III Enzyme Immunoassay | Predicate Device (K201938)LZI Fentanyl II Enzyme Immunoassay |
|---|---|---|
| Intended Use | The LZI Fentanyl III Enzyme Immunoassay is intended for the qualitative determination of fentanyl in human urine at the cutoff value of 1 ng/mL when calibrated against fentanyl. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. | The LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative detection of norfentanyl in human urine at the cutoff value of 5 ng/mL. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. |
| Analyte | Fentanyl | Norfentanyl |
| Cutoff | 1 ng/mL | 5 ng/mL |
| Matrix | Urine | same |
| Calibrator Level | 1 ng/mL | 5 ng/mL |
| Controls Level | 0.5 ng/mL and 1.5 ng/mL | 3.75 ng/mL and 6.25 ng/mL |
| Storage | 2-8 ºC until expiration date | same |
| Application Volume | R1: 65 µL; R2: 25 µL; Sample: 15 µL | R1: 120 µL; R2: 45 µL; Sample: 15 µL |
| Detection | Absorbance change measured spectrophotometrically at 340 nm. | same |
| User Environment | Clinical laboratories; Prescription use only | same |
| Mass Spectrometry Confirmation | Required to confirm preliminary positive analytical results | same |
| Platform Required | Automated clinical chemistry analyzer | same |
| Reagents Form | Liquid ready-to-use | same |
| Reagent Materials | Two (2) reagent system: Antibody/substrate reagent (R1) and enzyme labeled conjugates (R2) with sodium azide preservative | same |
Page 9
Lin-Zhi International, Inc.
Performance Characteristics Summary:
All 510(k) studies below were conducted on the Beckman Coulter AU5800 Analyzer
Precision: 1 ng/mL Cutoff
The assay was tested in qualitative (ΔOD, mAU) mode using a modified NCCLS-EP5 protocol. Fentanyl sample concentrations were prepared by spiking a fentanyl standard into a pool of negative human urine at the cutoff concentration and ±25%, ±50%, ±75%, and ±100% of the cutoff concentration.
The results shown below were obtained by testing all samples in replicates of two, two runs a day (one in the morning and one in the afternoon) for 22 days on one Beckman Coulter AU5800 automated clinical analyzer for a total of 88 replicates. Samples were evaluated against the cutoff calibrator in qualitative mode. One single lot of reagents, calibrators, and controls was used and stored at 2-8ºC when not in use.
Precision: 1 ng/mL Cutoff
Qualitative Positive/Negative Results:
| 1 ng/mL Cutoff Result: | Within Run (N=22) | Total Precision (N=88) | ||
|---|---|---|---|---|
| Fentanyl Concentration | % of Cutoff | Number of Determination | Immunoassay Result | Number of Determination |
| 0 ng/mL | 0% | 22 | 22 Negative | 88 |
| 0.25 ng/mL | 25% | 22 | 22 Negative | 88 |
| 0.5 ng/mL | 50% | 22 | 22 Negative | 88 |
| 0.75 ng/mL | 75% | 22 | 22 Negative | 88 |
| 1 ng/mL | 100% | 22 | 22 Positive | 88 |
| 1.25 ng/mL | 125% | 22 | 22 Positive | 88 |
| 1.5 ng/mL | 150% | 22 | 22 Positive | 88 |
| 1.75 ng/mL | 175% | 22 | 22 Positive | 88 |
| 2 ng/mL | 200% | 22 | 22 Positive | 88 |
Page 10
Lin-Zhi International, Inc.
Method Comparison - Clinical Samples:
A total of one hundred and fifty (150) unaltered clinical samples were tested with the LZI Fentanyl III Enzyme Immunoassay on the Beckman Coulter AU5800 automated clinical analyzer. Samples were evaluated against the cutoff calibrator in qualitative mode. All samples were tested in singlet.
All samples were confirmed by LC/MS for fentanyl concentrations. Samples were obtained by LZI and through collaboration with various clinical laboratories across the United States and Canada, including:
- APC Health (Tampa, Florida)
- Calgary Labs (Calgary, Canada)
- Carolina Liquid Chemistries Corporation (Greensboro, North Carolina)
- DTPM (Fort Payne, Alabama)
- Northwest Physicians Laboratories (Bellevue, Washington)
- Soloniuk Pain Clinic (Redding, California)
- Sterling Labs (Chicago, Illinois)
- TriCore Reference Laboratories (Santa Clara, California)
- University of California, San Francisco (San Francisco, California)
Qualitative Accuracy Study:
| FEN Results 1 ng/mL Cutoff | Negative by LC/MS analysis | < 50% of the cutoff concentration by LC/MS analysis | Near Cutoff Negative between 50% below the cutoff and the cutoff concentration by LC/MS analysis | Near Cutoff Positive between the cutoff and 50% above the cutoff concentration by LC/MS analysis | High Positive greater than 50% above the cutoff concentration by LC/MS analysis |
|---|---|---|---|---|---|
| Positive at or above the cutoff by EIA analysis | 0 | 20* | 12** | 22 | 61 |
| Negative below the cutoff by EIA analysis | 35 | 0 | 0 | 0 | 0 |
Page 11
Lin-Zhi International, Inc.
Method Comparison, continued:
Discrepant samples determined when comparing LC/MS fentanyl results with the LZI Fentanyl III EIA results on the Beckman Coulter AU5800 automated clinical analyzer.
| Sample # | LC/MS Fentanyl (ng/mL) | LC/MS Norfentanyl (ng/mL) | Pos/Neg Result | AU5800 EIA Qualitative Result (mAU) | Pos/Neg Result | Qualitative Cutoff Rate (mAU) |
|---|---|---|---|---|---|---|
| 36* | 0.0 | 1.5 | - | 83.7 | + | 19.3 |
| 37* | 0.0 | 4.1 | - | 172.3 | + | 19.3 |
| 38* | 0.1 | 1.1 | - | 57.3 | + | 19.3 |
| 39* | 0.1 | 6.7 | - | 203.9 | + | 19.3 |
| 40* | 0.1 | 4.1 | - | 234.2 | + | 19.3 |
| 41* | 0.1 | 4.4 | - | 192.7 | + | 19.3 |
| 42* | 0.1 | 6.1 | - | 134.5 | + | 19.3 |
| 43* | 0.1 | 5.3 | - | 150.6 | + | 19.3 |
| 44* | 0.1 | 4.4 | - | 104.9 | + | 19.3 |
| 45* | 0.2 | 4.8 | - | 206.2 | + | 19.3 |
| 46* | 0.2 | 1.9 | - | 214.7 | + | 19.3 |
| 47* | 0.2 | 2.0 | - | 151.3 | + | 19.3 |
| 48* | 0.2 | 5.1 | - | 101.1 | + | 19.3 |
| 49* | 0.2 | 5.0 | - | 206.2 | + | 19.3 |
| 50* | 0.2 | 39.9 | - | 213.6 | + | 22.1 |
| 51* | 0.2 | 10.0 | - | 155.3 | + | 19.3 |
| 52* | 0.3 | 2.4 | - | 171.4 | + | 19.3 |
| 53* | 0.3 | 9.7 | - | 253.1 | + | 19.3 |
| 54* | 0.3 | 4.2 | - | 166.2 | + | 19.3 |
| 55* | 0.3 | 5.4 | - | 162.5 | + | 19.3 |
| 56** | 0.6 | 2.2 | - | 185.6 | + | 19.3 |
| 57** | 0.6 | 362.5 | - | 225.1 | + | 19.3 |
| 58** | 0.7 | 15.9 | - | 228.2 | + | 22.1 |
| 59** | 0.7 | 101.7 | - | 246.8 | + | 22.1 |
| 60** | 0.7 | 167.1 | - | 260.2 | + | 19.3 |
| 61** | 0.8 | 30.2 | - | 314.7 | + | 19.3 |
| 62** | 0.8 | 2.8 | - | 45.0 | + | 22.1 |
| 63** | 0.8 | 0.5 | - | 108.2 | + | 19.3 |
| 64** | 0.8 | 2.8 | - | 134.8 | + | 19.3 |
| 65** | 0.9 | 15.8 | - | 224.3 | + | 19.3 |
| 66** | 0.9 | 106.7 | - | 318.7 | + | 22.1 |
| 67** | 0.9 | 12.0 | - | 258.4 | + | 19.3 |
* Values are discrepant below 50% of the cutoff concentration (0 ng/mL – 0.5 ng/mL fentanyl)
** Values are discrepant between 50% of the cutoff to the cutoff concentration (0.5 ng/mL – 1.0 ng/mL fentanyl)
All discrepant samples contained norfentanyl concentrations that contributed to the positive results.
Page 12
Lin-Zhi International, Inc.
Cross-reactivity
The cross-reactivity of various potentially interfering drugs were tested by spiking various concentrations of each substance into a pool of negative human urine and then evaluated against the assay's calibration curve in qualitative mode. All samples were tested in duplicates.
The table below lists the concentration of each test compound that gave a response approximately equivalent to that of the cutoff calibrator (as positive) or the maximal concentration of the compound tested that gave a response below the response of the cutoff calibrator (as negative). Compounds tested at high concentration (100,000 ng/mL) with results below the cutoff value were listed as Not Detected (ND).
Fentanyl and Metabolites:
| Compound | Test Concentration (ng/mL) | Qualitative Result (mAU) | Qualitative Cutoff Rate (mAU) | % Cross-reactivity |
|---|---|---|---|---|
| Fentanyl | 1.0 | 26.1 | 17.9 | 100.00 % |
| Norfentanyl | 2.5 | 24.4 | 17.9 | 40.00 % |
Structurally Related Compounds:
| Compound | Test Concentration (ng/mL) | Qualitative Result (mAU) | Qualitative Cutoff Rate (mAU) | % Cross-reactivity |
|---|---|---|---|---|
| 4-Fluoro-isobutyryl fentanyl | 25 | 22.1 | 20.2 | 4.00% |
| 9-Hydroxy risperidone | 100,000 | -19.8 | 20.2 | ND |
| Acetyl fentanyl | 4 | 22.8 | 20.2 | 25.00% |
| Acetyl norfentanyl | 100 | 22.1 | 20.2 | 1.00% |
| Acryl fentanyl | 1 | 36.1 | 21.1 | 100.00% |
| Alfentanil | 100,000 | -18.5 | 20.2 | ND |
| Benzyl fentanyl | 2 | 20.7 | 20.2 | 50.00% |
| Butyryl fentanyl | 1 | 24.5 | 21.1 | 100.00% |
| Butyryl norfentanyl | 40 | 26.1 | 17.9 | 2.50% |
| Carfentanil oxalate | 100,000 | 2.7 | 20.2 | ND |
| (±)cis-3-methyl fentanyl | 4.5 | 34.8 | 20.2 | 22.22% |
| Cyclopropyl norfentanyl | 15 | 21.3 | 17.9 | 6.67% |
| Despropionyl fentanyl (4-ANPP) | 100,000 | -2.1 | 20.2 | ND |
| Furanyl fentanyl | 2.5 | 25.5 | 21.1 | 40.00% |
| Furanyl norfentanyl | 150 | 22.4 | 17.9 | 0.67% |
| β-Hydroxyfentanyl | 1.1 | 30.4 | 17.9 | 90.91% |
| (±)-β-hydroxythiofentanyl | 1 | 25.6 | 21.1 | 100.00% |
| Isobutyryl fentanyl | 12.5 | 21.9 | 17.9 | 8.00% |
| Isobutyryl norfentanyl | 390 | 24.5 | 17.9 | 0.26% |
| Methoxyacetyl fentanyl | 1 | 30.1 | 21.1 | 100.00% |
Page 13
Lin-Zhi International, Inc.
Structurally Related Compounds, continued:
| Compound | Test Concentration (ng/mL) | Qualitative Result (mAU) | Qualitative Cutoff Rate (mAU) | % Cross-reactivity |
|---|---|---|---|---|
| MT-45 | 100,000 | -22.3 | 20.2 | ND |
| N-benzyl furanyl norfentanyl | 0.75 | 20.4 | 20.2 | 133.33% |
| N-benzyl para-fluoro norfentanyl | 1 | 32.2 | 20.2 | 100.00% |
| Norcarfentanil oxalate | 100,000 | -6.5 | 20.2 | ND |
| Ocfentanil | 1 | 27.4 | 21.1 | 100.00% |
| Para-fluorobutyryl fentanyl | 1.2 | 19.4 | 17.9 | 83.33% |
| Para-fluoro fentanyl | 1 | 32.3 | 21.1 | 100.00% |
| Para-methoxy-butyryl fentanyl | 1 | 24.6 | 17.9 | 100.00% |
| Remifentanil | 100,000 | -5.5 | 28.3 | ND |
| Risperidone | 100,000 | -40.5 | 20.2 | ND |
| Sufentanil | 1,000 | 23.2 | 20.2 | 0.10% |
| Thienyl fentanyl | 0.75 | 24.9 | 17.9 | 133.33% |
| Thiofentanyl | 0.50 | 21.6 | 20.2 | 200.00% |
| Trans-3-methyl fentanyl | 2 | 18.9 | 17.9 | 50.00% |
| Trazodone | 100,000 | -7.0 | 20.2 | ND |
| U-47700 | 100,000 | -22.6 | 20.2 | ND |
| Valeryl fentanyl | 50 | 21.3 | 20.2 | 2.00% |
| Ortho-methyl fentanyl | 3.5 | 27.5 | 20.2 | 28.57% |
Page 14
Lin-Zhi International, Inc.
Cross-reactivity, continued:
Structurally Unrelated Pharmacological Compounds:
| Compound | Test Concentration (ng/mL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| (1S,2S)-(+)Pseudoephedrine | 100,000 | Neg | Neg | Pos |
| 6-Acetylmorphine | 100,000 | Neg | Neg | Pos |
| 7-Hydroxymitragynine | 100,000 | Neg | Neg | Pos |
| Acetaminophen | 100,000 | Neg | Neg | Pos |
| Acetylsalicylic acid | 100,000 | Neg | Neg | Pos |
| AH 7921 | 100,000 | Neg | Neg | Pos |
| Alprazolam | 100,000 | Neg | Neg | Pos |
| Amitriptyline | 100,000 | Neg | Neg | Pos |
| Amlodine besylate | 100,000 | Neg | Neg | Pos |
| Amobarbital | 100,000 | Neg | Neg | Pos |
| Amoxicillin | 100,000 | Neg | Neg | Pos |
| d-Amphetamine | 100,000 | Neg | Neg | Pos |
| Aripiprazole | 100,000 | Neg | Neg | Pos |
| Atorvastatin | 100,000 | Neg | Neg | Pos |
| Benzoylecgonine | 100,000 | Neg | Neg | Pos |
| Bisoprolol | 100,000 | Neg | Neg | Pos |
| Bromazepam | 100,000 | Neg | Neg | Pos |
| Buprenorphine | 100,000 | Neg | Neg | Pos |
| Buprenorphine glucuronide | 100,000 | Neg | Neg | Pos |
| Bupropion | 100,000 | Neg | Neg | Pos |
| Butalbital | 100,000 | Neg | Neg | Pos |
| Caffeine | 100,000 | Neg | Neg | Pos |
| Cannabidiol | 100,000 | Neg | Neg | Pos |
Page 15
Lin-Zhi International, Inc.
Structurally Unrelated Pharmacological Compounds, continued:
| Compound | Test Concentration (ng/mL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| Carbamazepine | 100,000 | Neg | Neg | Pos |
| Carisoprodol | 100,000 | Neg | Neg | Pos |
| Cetirizine | 100,000 | Neg | Neg | Pos |
| Chlordiazepoxide | 100,000 | Neg | Neg | Pos |
| Chlorpheniramine | 100,000 | Neg | Neg | Pos |
| Chlorpromazine | 100,000 | Neg | Neg | Pos |
| Clobazam | 100,000 | Neg | Neg | Pos |
| Clomipramine | 100,000 | Neg | Neg | Pos |
| Clonazepam | 100,000 | Neg | Neg | Pos |
| Cocaine | 100,000 | Neg | Neg | Pos |
| Codeine | 100,000 | Neg | Neg | Pos |
| Cotinine | 100,000 | Neg | Neg | Pos |
| Cyclobenzaprine | 100,000 | Neg | Neg | Pos |
| Desipramine | 100,000 | Neg | Neg | Pos |
| Dextromethorphan | 20,000 | Neg | Pos | Pos |
| Diazepam | 100,000 | Neg | Neg | Pos |
| Diphenhydramine | 100,000 | Neg | Neg | Pos |
| Doxepin | 100,000 | Neg | Neg | Pos |
| Doxylamine | 100,000 | Neg | Neg | Pos |
| Duloxetine | 100,000 | Neg | Neg | Pos |
| EDDP | 100,000 | Neg | Neg | Pos |
| Ecgonine | 100,000 | Neg | Neg | Pos |
| Ecgonine methyl ester | 100,000 | Neg | Neg | Pos |
| EMDP | 100,000 | Neg | Neg | Pos |
| Ephedrine | 100,000 | Neg | Neg | Pos |
| Fexofenadine | 100,000 | Neg | Neg | Pos |
| Flunitrazepam | 5000 | Neg | Neg | Pos |
| Fluoxetine | 100,000 | Neg | Neg | Pos |
| Fluphenazine | 100,000 | Neg | Neg | Pos |
| Flurazepam | 100,000 | Neg | Neg | Pos |
| Furosemide | 100,000 | Neg | Neg | Pos |
| Gabapentin | 100,000 | Neg | Neg | Pos |
| Haloperidol | 100,000 | Neg | Neg | Pos |
| Heroin | 100,000 | Neg | Neg | Pos |
| Hexobarbital | 100,000 | Neg | Neg | Pos |
| Hydrochlorothiazide | 100,000 | Neg | Neg | Pos |
| Hydrocodone | 100,000 | Neg | Neg | Pos |
| Hydromorphone | 100,000 | Neg | Neg | Pos |
Page 16
Lin-Zhi International, Inc.
Structurally Unrelated Pharmacological Compounds, continued:
| Compound | Test Concentration (ng/mL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| Ibuprofen | 100,000 | Neg | Neg | Pos |
| Imipramine | 100,000 | Neg | Neg | Pos |
| Ketamine | 100,000 | Neg | Neg | Pos |
| Labetalol | 100,000 | Neg | Neg | Pos |
| Lamotrigine | 100,000 | Neg | Neg | Pos |
| Levorphanol | 100,000 | Neg | Neg | Pos |
| Lidocaine | 100,000 | Neg | Neg | Pos |
| Lisinopril | 100,000 | Neg | Neg | Pos |
| Loratadine | 100,000 | Neg | Neg | Pos |
| Lormetazepam | 100,000 | Neg | Neg | Pos |
| Losartan | 100,000 | Neg | Neg | Pos |
| LSD | 100,000 | Neg | Neg | Pos |
| Maprotiline | 100,000 | Neg | Neg | Pos |
| mCPP | 100,000 | Neg | Neg | Pos |
| MDA | 100,000 | Neg | Neg | Pos |
| MDEA | 100,000 | Neg | Neg | Pos |
| MDMA | 100,000 | Neg | Neg | Pos |
| Meperidine | 100,000 | Neg | Neg | Pos |
| Mephobarbital | 100,000 | Neg | Neg | Pos |
| Meprobamate | 100,000 | Neg | Neg | Pos |
| Metformin | 100,000 | Neg | Neg | Pos |
| Methadone | 100,000 | Neg | Neg | Pos |
| d-Methamphetamine | 100,000 | Neg | Neg | Pos |
| Methapyrilene | 100,000 | Neg | Neg | Pos |
| Methaqualone | 100,000 | Neg | Neg | Pos |
| Methylphenidate | 100,000 | Neg | Neg | Pos |
| Metoprolol | 100,000 | Neg | Neg | Pos |
| Metronidazole | 100,000 | Neg | Neg | Pos |
| Minocycline | 100,000 | Neg | Neg | Pos |
| Mirtazapine | 100,000 | Neg | Neg | Pos |
| Morphine | 100,000 | Neg | Neg | Pos |
| Morphine-3-glucuronide | 100,000 | Neg | Neg | Pos |
| Nalmefene | 100,000 | Neg | Neg | Pos |
| Naloxone | 100,000 | Neg | Neg | Pos |
| Naltrexone | 100,000 | Neg | Neg | Pos |
| Naproxen | 100,000 | Neg | Neg | Pos |
| Nicotine | 100,000 | Neg | Neg | Pos |
Page 17
Lin-Zhi International, Inc.
Structurally Unrelated Pharmacological Compounds, continued:
| Compound | Test Concentration (ng/mL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| Nitrazepam | 100,000 | Neg | Neg | Pos |
| Norbuprenorphine | 100,000 | Neg | Neg | Pos |
| Norcodeine | 100,000 | Neg | Neg | Pos |
| Norketamine | 100,000 | Neg | Neg | Pos |
| Normeperidine | 100,000 | Neg | Neg | Pos |
| Normorphine | 100,000 | Neg | Neg | Pos |
| Norquetiapine | 100,000 | Neg | Neg | Pos |
| Nortriptyline | 100,000 | Neg | Neg | Pos |
| Olanzapine | 100,000 | Neg | Neg | Pos |
| Omeprazole | 100,000 | Neg | Neg | Pos |
| Oxazepam | 100,000 | Neg | Neg | Pos |
| Oxycodone | 100,000 | Neg | Neg | Pos |
| Oxymorphone | 100,000 | Neg | Neg | Pos |
| Paroxetine | 100,000 | Neg | Neg | Pos |
| Pentazocine | 100,000 | Neg | Neg | Pos |
| Pentobarbital | 100,000 | Neg | Neg | Pos |
| Perphenazine | 100,000 | Neg | Neg | Pos |
| Phencyclidine | 100,000 | Neg | Neg | Pos |
| Phenobarbital | 100,000 | Neg | Neg | Pos |
| Phentermine | 100,000 | Neg | Neg | Pos |
| Propoxyphene | 100,000 | Neg | Neg | Pos |
| Quetiapine | 100,000 | Neg | Neg | Pos |
| Quinidine | 100,000 | Neg | Neg | Pos |
| Ranitidine | 100,000 | Neg | Neg | Pos |
| Ritalinic acid | 100,000 | Neg | Neg | Pos |
| Phenytoin | 100,000 | Neg | Neg | Pos |
| Salbutamol | 100,000 | Neg | Neg | Pos |
| Salicylic acid | 100,000 | Neg | Neg | Pos |
| Secobarbital | 100,000 | Neg | Neg | Pos |
| Sertraline | 100,000 | Neg | Neg | Pos |
| Sildenafil | 100,000 | Neg | Neg | Pos |
| Tapentadol | 100,000 | Neg | Neg | Pos |
| Temazepam | 100,000 | Neg | Neg | Pos |
| THC | 100,000 | Neg | Neg | Pos |
| Thebaine | 100,000 | Neg | Neg | Pos |
| Theophylline | 100,000 | Neg | Neg | Pos |
| Thioridazine | 100,000 | Neg | Neg | Pos |
| l-Thyroxine | 100,000 | Neg | Neg | Pos |
Page 18
Lin-Zhi International, Inc.
Structurally Unrelated Pharmacological Compounds, continued:
| Compound | Test Concentration (ng/mL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| Tilidine | 100,000 | Neg | Neg | Pos |
| Tramadol | 100,000 | Neg | Neg | Pos |
| Trazadone | 100,000 | Neg | Neg | Pos |
| Triazolam | 100,000 | Neg | Neg | Pos |
| Trimipramine | 100,000 | Neg | Neg | Pos |
| Valproic Acid | 100,000 | Neg | Neg | Pos |
| Verapamil | 100,000 | Neg | Neg | Pos |
| Venlafaxine | 100,000 | Neg | Neg | Pos |
| Ziprasidone | 100,000 | Neg | Neg | Pos |
| Zolpidem | 100,000 | Neg | Neg | Pos |
| Zopiclone | 100,000 | Neg | Neg | Pos |
It is possible that other substances and/or factors not listed above may interfere with the test and cause false results, e.g., technical or procedural errors
The following structurally unrelated compound which showed interference at ±50% of the cutoff concentration was then tested at the highest concentration that did not cause a discrepant result. The result is summarized in the following table:
| Compound | Spiked [ ] (ng/mL) | Spiked Fentanyl Concentration | ||
|---|---|---|---|---|
| 0 ng/mL (ng/mL) | 0.5 ng/mL Control (ng/mL) | 1.5 ng/mL Control (ng/mL) | ||
| Dextromethorphan | 15,000 | Neg | Neg | Pos |
Page 19
Lin-Zhi International, Inc.
Endogenous and Preservative Compound Interference:
Endogenous and Preservative compounds were spiked into pooled negative human urine to desired concentrations. These solutions were then split into three portions; one without fentanyl, and the remaining two that were further spiked with fentanyl standards to a final fentanyl concentration of 0.5 ng/mL or 1.5 ng/mL (as negative or positive controls, ±50% of the cutoff concentration, respectively). Samples were then evaluated against the assay's calibration curve in qualitative mode. All samples were tested in duplicates.
Interference was observed with Boric Acid at 1% w/v. No other significant cross-reactivity was observed.
| Interfering Substance | Concentration of Compound (mg/dL) | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|---|
| Acetone | 1,000 | Neg | Neg | Pos |
| Ascorbic acid | 500 | Neg | Neg | Pos |
| Bilirubin | 2 | Neg | Neg | Pos |
| Biotin | 2 | Neg | Neg | Pos |
| Boric acid | 1,000 | Neg | Neg | Neg |
| Calcium chloride | 300 | Neg | Neg | Pos |
| Citric acid | 200 | Neg | Neg | Pos |
| Creatinine | 500 | Neg | Neg | Pos |
| Ethanol | 1,000 | Neg | Neg | Pos |
| Galactose | 10 | Neg | Neg | Pos |
| γ-Globulin | 500 | Neg | Neg | Pos |
| Glucose | 3,000 | Neg | Neg | Pos |
| Hemoglobin | 300 | Neg | Neg | Pos |
| Human urine (pooled) | N/A | Neg | Neg | Pos |
| Human serum albumin | 500 | Neg | Neg | Pos |
| β-Hydroxybutyric acid | 100 | Neg | Neg | Pos |
| Oxalic acid | 100 | Neg | Neg | Pos |
| Potassium chloride | 1,000 | Neg | Neg | Pos |
| Riboflavin | 7.5 | Neg | Neg | Pos |
| Sodium azide | 1,000 | Neg | Neg | Pos |
| Sodium chloride | 1,000 | Neg | Neg | Pos |
| Sodium fluoride | 1,000 | Neg | Neg | Pos |
| Sodium phosphate | 300 | Neg | Neg | Pos |
| Urea | 6,000 | Neg | Neg | Pos |
| Uric acid | 10 | Neg | Neg | Pos |
| Urine-based calibrator buffer | N/A | Neg | Neg | Pos |
Page 20
Lin-Zhi International, Inc.
Specific Gravity Interference:
Samples ranging in specific gravity from 1.000 to 1.030 were split into three portions each and either left un-spiked or further spiked to a final fentanyl concentration of either 0.5 ng/mL or 1.5 ng/mL (as negative or positive controls, ±50% of the cutoff concentration, respectively). These samples were then evaluated in qualitative mode. No interference was observed.
| Specific Gravity Value | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|
| 1.000 | Neg | Neg | Pos |
| 1.003 | Neg | Neg | Pos |
| 1.005 | Neg | Neg | Pos |
| 1.008 | Neg | Neg | Pos |
| 1.010 | Neg | Neg | Pos |
| 1.011 | Neg | Neg | Pos |
| 1.013 | Neg | Neg | Pos |
| 1.015 | Neg | Neg | Pos |
| 1.018 | Neg | Neg | Pos |
| 1.020 | Neg | Neg | Pos |
| 1.022 | Neg | Neg | Pos |
| 1.023 | Neg | Neg | Pos |
| 1.025 | Neg | Neg | Pos |
| 1.030 | Neg | Neg | Pos |
Page 21
Lin-Zhi International, Inc.
pH Interference:
Negative urine and urine spiked with fentanyl to the final fentanyl concentration of either 0.5 ng/mL or 1.5 ng/mL (as negative or positive controls, ±50% of the cutoff concentration, respectively) were adjusted to the following pH levels and tested by the assay. The pH adjusted solutions were evaluated in qualitative mode.
No major interference was observed between pH 3 to pH 11. Results are summarized in the following table:
| Interfering Substance | 0 ng/mL Fentanyl | -50% Fentanyl Cutoff (0.5 ng/mL) | +50% Fentanyl Cutoff (1.5 ng/mL) |
|---|---|---|---|
| pH 3 | Neg | Neg | Pos |
| pH 4 | Neg | Neg | Pos |
| pH 5 | Neg | Neg | Pos |
| pH 6 | Neg | Neg | Pos |
| pH 7 | Neg | Neg | Pos |
| pH 8 | Neg | Neg | Pos |
| pH 9 | Neg | Neg | Pos |
| pH 10 | Neg | Neg | Pos |
| pH 11 | Neg | Neg | Pos |
Page 22
Lin-Zhi International, Inc.
Conclusion:
The information provided in this pre-market notification demonstrates that the LZI Fentanyl III Enzyme Immunoassay is substantially equivalent to the legally marketed predicate device for its general intended use. Substantial equivalence was demonstrated through comparison of intended use and physical properties to the commercially available predicate device as confirmed by chromatography/mass spectrometry (GC/MS or LC/MS), an independent analytical method. The information supplied in this pre-market notification provides reasonable assurance that the LZI Fentanyl III Enzyme Immunoassay is safe and effective for its stated intended use.
§ 862.3650 Opiate test system.
(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (e.g., programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).