(25 days)
The LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative determination 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.
The LZI Fentanyl II Enzyme Immunoassay is a homogeneous enzyme immunoassay with readyto-use liguid reagents. The assay is based on competition between drug in the sample and 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 drug in the sample, fentanyl-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. On the other hand, when free drug is present in the sample, antibody would bind to free drug; the unbound fentanyllabeled 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 II Enzyme Immunoassay is a kit comprised of two reagents, an R1 and R2 which are bottled separately but sold together within the kit. The LZI Fentanyl II Enzyme Immunoassay is traceable to a commercially available fentanyl standard.
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 LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative determination of norfentanyl in human urine at a cutoff value of 5 ng/mL. The device provides a preliminary analytical result, and a more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used for a confirmed analytical result.
Here's an analysis of the acceptance criteria and the study that proves the device meets them:
1. A table of acceptance criteria and the reported device performance:
| Acceptance Criteria Category | Specific Criteria | Reported Device Performance |
|---|---|---|
| Precision | Ability to consistently produce correct qualitative results (Negative/Positive) for samples with concentrations at various percentages around the 5 ng/mL cutoff (0%, 25%, 50%, 75%, 100%, 125%, 150%, 175%, 200%). | Within Run (N=22): All 0%, 25%, 50%, 75% negative samples were Negative. All 125%, 150%, 175%, 200% positive samples were Positive. 100% cutoff samples were 13 Neg/9 Pos (indicating some variability around the cutoff, which is expected for qualitative assays at the cutoff). Total Precision (N=88): All 0%, 25%, 50%, 75% negative samples were Negative. All 125%, 150%, 175%, 200% positive samples were Positive. 100% cutoff samples were 59 Neg/29 Pos. |
| Method Comparison (Qualitative Accuracy) | Comparison of EIA results with LC/MS confirmation for clinical samples, especially focusing on false positives and false negatives within different concentration ranges relative to the cutoff. Acceptable levels of agreement and defined explanations for discrepancies. | Overall Accuracy: 20/20 negative samples (by LC/MS) were EIA Negative. 19/19 samples <50% of cutoff (by LC/MS) were EIA Negative. 2/10 samples Near Cutoff Negative (by LC/MS) were EIA Negative, while 8 were EIA Positive. 10/10 Near Cutoff Positive samples (by LC/MS) were EIA Positive. 40/40 High Positive samples (by LC/MS) were EIA Positive. Discrepant Samples: 9 samples with LC/MS norfentanyl concentrations between 1.5 ng/mL and 4.2 ng/mL (below the 5 ng/mL cutoff) were reported as EIA Positive. These were attributed to "levels of fentanyl that contributed to the false positive result" (cross-reactivity from fentanyl itself). |
| Cross-reactivity | Identification of compounds (structurally related, unrelated pharmacological, endogenous, preservatives) that might interfere with the assay, and quantification of their cross-reactivity. Acceptable cross-reactivity levels (e.g., "Not Detected" or specific percentage) for various substances. | Fentanyl and Metabolites: Fentanyl showed 131.58% cross-reactivity at 3.8 ng/mL. Norfentanyl showed 100.00% cross-reactivity at 5 ng/mL (as expected). Many other fentanyl analogs and metabolites showed significant cross-reactivity (e.g., Cyclopropyl Fentanyl 156.25% at 3.2 ng/mL). Structurally Unrelated Compounds: Most tested at 100,000 ng/mL showed "ND" (Not Detected). Dextromethorphan interfered at 40,000 ng/mL, showing 0.01% cross-reactivity and causing a positive result at the -25% Norfentanyl Cutoff (3.75 ng/mL). Endogenous & Preservative Compounds: Most showed no interference. Boric Acid showed interference at 1000 mg/dL, causing a negative result even at the +50% Norfentanyl Cutoff (7.5 ng/mL). |
| Specific Gravity Interference | No qualitative result interference across a range of specific gravity values (1.000 to 1.027) for negative, -25% cutoff, and +25% cutoff samples. | No interference was observed across the tested specific gravity range (1.000 to 1.027) for negative, -25% cutoff, and +25% cutoff samples. |
| pH Interference | No qualitative result interference across a range of pH values (pH 3 to pH 11) for negative, -25% cutoff, and +25% cutoff samples. | No major interference was observed between pH 3 to pH 11 for negative, -25% cutoff, and +25% cutoff samples. |
Important Note: The document focuses on demonstrating substantial equivalence to the predicate device and does not explicitly state numerical "acceptance criteria" for precision or accuracy, but rather reports the performance of the device against established test designs (e.g., NCCLS-EP5 for precision) and confirms qualitative results against a gold standard (LC/MS). The tables demonstrate the device's performance characteristics.
2. Sample size used for the test set and the data provenance:
- Precision Test Set (Qualitative): 88 replicates per concentration level (for 9 concentration levels from 0 to 200% of cutoff), making a total of 792 individual tests. These were prepared by spiking known norfentanyl standards into a pool of negative human urine.
- Method Comparison - Clinical Samples (Qualitative Accuracy): 100 unaltered clinical samples.
- Data Provenance: Samples were collected by Lin-Zhi International, Inc. and from various clinical labs: University of California, San Francisco (UCSF) (San Francisco, CA), Northwest Physicians Labs (NWPL) (Bellevue, Washington), Soloniuk Pain Clinic (Redding, CA), and Calgary Labs (Calgary, Canada). The study represents a retrospective analysis of these collected samples.
- Cross-reactivity, Endogenous, Preservative, Specific Gravity, and pH Interference Tests: Samples were prepared by spiking various compounds into pooled negative human urine or by adjusting the urine's properties. These were not external clinical samples in the same way as the method comparison set.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g., radiologist with 10 years of experience):
The ground truth for clinical samples (method comparison) was established using LC/MS (Liquid Chromatography/Mass Spectrometry) analysis. LC/MS is a highly specific and sensitive analytical technique, often considered the gold standard for drug quantification in toxicology. This method does not typically involve human experts establishing a subjective "ground truth" but rather relies on the analytical results of the instrument. The document does not specify the qualifications of the personnel performing the LC/MS analysis.
For the other test sets (precision, cross-reactivity, interference), the ground truth was established by the known concentrations of spiked compounds or the known properties of the prepared urine samples.
4. Adjudication method (e.g., 2+1, 3+1, none) for the test set:
There was no adjudication method involving multiple human readers described for establishing the ground truth or interpreting the results.
- For the precision study, the qualitative results (Negative/Positive) were derived directly from the automated clinical analyzer's response compared to the cutoff calibrator.
- For the method comparison (clinical samples), the ground truth was solely determined by the LC/MS reference method.
- For interference studies, the outcomes were based on the instrument's qualitative determination relative to the cutoff.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
No MRMC comparative effectiveness study was done. This device is an in vitro diagnostic (IVD) immunoassay performed on an automated clinical chemistry analyzer, not an AI-assisted diagnostic imaging or interpretative tool that would involve human readers.
6. If a standalone (i.e. algorithm only without human-in-the loop performance) was done:
Yes, the performance studies described are essentially standalone (algorithm/device-only performance). The LZI Fentanyl II Enzyme Immunoassay operates on an automated clinical analyzer (Beckman Coulter AU480), and its performance metrics (precision, qualitative accuracy against LC/MS, cross-reactivity, interference) are determined directly from the instrument's output based on its chemical reactions and detection system, without human interpretation in the diagnostic process.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
The primary ground truth used was analytical reference methods, specifically:
- LC/MS (Liquid Chromatography/Mass Spectrometry) for confirming the concentration of norfentanyl in clinical urine samples. This is considered an objective gold standard for drug testing.
- Known concentrations/properties for prepared spiked samples (for precision, cross-reactivity, and interference studies).
8. The sample size for the training set:
This document describes a premarket notification for an immunoassay kit, which is a chemical and enzymatic assay, not a machine learning or AI algorithm. Therefore, there is no "training set" in the context of data used to train an AI model. The device's performance is based on its reagents' chemical properties and reaction kinetics.
9. How the ground truth for the training set was established:
As there is no training set for an AI model, this question is not applicable. The device's performance is intrinsically linked to the validated formulation of its reagents and the instrument's analytical capabilities, which are established through general scientific and manufacturing practices, not data-driven AI training.
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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
August 7, 2020
Lin-Zhi International, Inc. Bernice Lin VP Operations 2945 Oakmead Village Court Santa Clara, CA 95051
Re: K201938
Trade/Device Name: LZI Fentanyl II Enzyme Immunoassay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: DJG Dated: July 8, 2020 Received: July 13, 2020
Dear Bernice Lin:
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 (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 located 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.
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
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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-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 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 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,
Marianela Perez-Torres, Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K201938
Device Name LZI Fentanyl II Enzyme Immunoassay
Indications for Use (Describe)
The LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative determination 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.
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
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510(k) Summary of Safety and Effectiveness
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
510(k) Number
Prepared On
July 2, 2020
Introduction
According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.
Submitter Name, Address, and Contact:
Lin-Zhi International, Inc. 2945 Oakmead Village Court Santa Clara, CA 95051 Phone: (408) 970-8811 (408) 970-9030 Fax: e-mail: bclin@lin-zhi.com
Contact: Bernice Lin, Ph.D. VP Operations
Device Name and Classification
| Classification Name: | Enzyme Immunoassay, OpiatesClass II, DJG (91 Toxicology),21 CFR 862.3650 |
|---|---|
| Common Name: | Homogeneous Fentanyl Enzyme Immunoassay |
| Proprietary Name: | LZI Fentanyl II Enzyme Immunoassay |
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Legally Marketed Predicate Device(s)
The LZI Fentanyl II Enzyme Immunoassay is substantially equivalent to the LZI Fentanyl Enzyme Immunoassay (K181159) manufactured by Lin-Zhi International, Inc. The LZI Fentanyl II Enzyme Immunoassay is identical or similar to its predicate in terms of intended use, method principle, device components, and clinical performance.
Device Description
The LZI Fentanyl II Enzyme Immunoassay is a homogeneous enzyme immunoassay with readyto-use liguid reagents. The assay is based on competition between drug in the sample and 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 drug in the sample, fentanyl-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. On the other hand, when free drug is present in the sample, antibody would bind to free drug; the unbound fentanyllabeled 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 II Enzyme Immunoassay is a kit comprised of two reagents, an R1 and R2 which are bottled separately but sold together within the kit. The LZI Fentanyl II Enzyme Immunoassay is traceable to a commercially available fentanyl standard.
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.
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Intended Use
The LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative determination 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 chromatograpy 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.
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Comparison to Predicate Device
The LZI Fentanyl II Enzyme Immunoassay is substantially equivalent to the LZI Fentanyl Enzyme Immunoassay which was cleared by the FDA under the premarket notification K181159 for its stated intended use.
The following table compares the LZI Fentanyl II Enzyme Immunoassay with the predicate device.
| DeviceCharacteristics | Subject Device | Predicate Device (K181159) |
|---|---|---|
| Intended Use | The LZI Fentanyl II Enzyme Immunoassay is intended for the qualitative determination of norfentanyl in human urine at the cutoff value of 5 ng/mL when calibrated against norfentanyl. The assay is designed for professional use with a number of automated clinical chemistry analyzers.This assay provides a rapid screening procedure for determining the presence of norfentanyl in urine. The assay provides only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography/mass spectrometry (GC/MS or LC/MS) is the preferred confirmatory method. 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 Enzyme Immunoassay is intended for the qualitative determination of norfentanyl in human urine at the cutoff value of 5 ng/mL when calibrated against norfentanyl. The assay is designed for professional use with a number of automated clinical chemistry analyzers.This assay provides a rapid screening procedure for determining the presence of norfentanyl in urine. The assay provides only a preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography/mass spectrometry (GC/MS or LC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. |
| Analyte | norfentanyl | norfentanyl |
| Cutoff | 5 ng/mL | 5 ng/mL |
| Matrix | Urine | Urine |
| CalibratorLevel | 5 ng/mL | 5 ng/mL |
| Controls Level | 3.75 ng/mL and 6.25 ng/mL | 3.75 ng/mL and 6.25 ng/mL |
| Storage | 2-8 °C until expiration date | 2-8 °C until expiration date |
| Instrumentused forperformance | Beckman Coulter® AU480 automatedclinical analyzer | Beckman Coulter® AU680 automatedclinical analyzer |
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Performance Characteristics Summary:
All 510(k) studies below were conducted on the Beckman Coulter® AU480 Analyzer
Precision: 5 ng/mL Cutoff
The assay was tested in qualitative (AOD, mAU) mode using a modified NCCLS-EP5 protocol. Norfentanyl sample concentrations were prepared by spiking a norfentanyl standard into a pool of negative human urine at concentrations ±25 %, ±75 %, and ±100 % of the cutoff concentration.
Results shown below were obtained by testing all samples in replicate of two, two runs a day (one in the morning and one in the afternoon) for 22 days on one Beckman Coulter® AU480 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 were used and stored at 2-8℃ when not in use.
| NorfentanylConcentration | Within Run (N=22) | Total Precision (N=88) | ||||
|---|---|---|---|---|---|---|
| Mean | SD | %CV | Mean | SD | % CV | |
| 0 ng/mL | 0.0 | 0.0 | N/A | 0.0 | 0.2 | N/A |
| 1.25 ng/mL | 15.5 | 3.0 | 20.2 % | 15.5 | 4.1 | 26.6 % |
| 2.5 ng/mL | 35.1 | 2.9 | 8.4 % | 35.1 | 3.5 | 10.0 % |
| 3.75 ng/mL | 58.8 | 3.0 | 5.1 % | 58.8 | 3.8 | 6.5 % |
| 5 ng/mL | 81.0 | 3.3 | 4.1 % | 81.0 | 3.7 | 4.6 % |
| 6.25 ng/mL | 105.5 | 2.7 | 2.5 % | 105.5 | 3.9 | 3.7 % |
| 7.5 ng/mL | 128.0 | 2.9 | 2.3 % | 128.0 | 3.5 | 2.8 % |
| 8.75 ng/mL | 150.1 | 3.2 | 2.2 % | 150.1 | 4.2 | 2.8 % |
| 10 ng/mL | 171.3 | 4.3 | 2.5 % | 171.3 | 5.2 | 3.0 % |
Precision: 5 ng/mL Cutoff Precision: Qualitative, results in ΔΟD (mAU)
Oualitative Positive/Negative Results:
| 5 ng/mL Cutoff Result: | Within Run (N=22) | Total Precision (N=88) | |||
|---|---|---|---|---|---|
| Norfentanyl | % of Cutoff | Number of | Immunoassay | Number of | Immunoassay |
| Concentration | Determination | Result | Determination | Result | |
| 0 ng/mL | 0.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 1.25 ng/mL | 25.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 2.5 ng/mL | 50.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 3.75 ng/mL | 75.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 5 ng/mL | 100.0 % | 22 | 13 Neg/9 Pos | 88 | 59 Neg/29 Pos |
| 6.25 ng/mL | 125.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 7.5 ng/mL | 150.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 8.75 ng/mL | 175.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 10 ng/mL | 200.0 % | 22 | 22 Positive | 88 | 88 Positive |
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Beckman Coulter® AU480 Analyzer
Method Comparison - Clinical Samples:
A total of one hundred (100) unaltered clinical samples were tested with the LZI Fentanyl II Enzyme Immunoassay on the Beckman Coulter® AU480 automated clinical analyzer. Samples were evaluated against the cutoff calibrator in qualitative mode. All samples were tested in singlet.
All samples were confirmed with LC/MS for norfentanyl concentrations. Samples were collected by Lin-Zhi International, Inc. and from various clinical labs including University of California, San Francisco (UCSF) (San Francisco, CA), Northwest Physicians Labs (NWPL) (Bellevue, Washington), Soloniuk Pain Clinic (Redding, CA), and Calgary Labs (Calgary, Canada).
Qualitative Accuracy Study:
| NFEN Results5 ng/mL Cutoff | Negativeby LC/MSanalysis | < 50 % of thecutoffconcentration byLC/MS analysis | Near CutoffNegative(Between 50 %below the cutoffand the cutoffconcentration byLC/MS analysis) | Near CutoffPositive (Betweenthe cutoff and 50% above the cutoffconcentration byLC/MS analysis) | High Positive(Greater than 50% above thecutoffconcentration byLC/MS analysis) |
|---|---|---|---|---|---|
| Positive(at or above thecutoff by EIAanalysis) | 0 | 1* | 8* | 10 | 40 |
| Negative(below the cutoffby EIA analysis) | 20 | 19 | 2 | 0 | 0 |
Discrepant samples determined when comparing LC/MS norfentanyl results with the LZI Fentanyl II EIA results on the Beckman Coulter® AU480 automated clinical analyzer.
| Sample # | LC/MS Norfentanyl (ng/mL) | Pos/ Neg Result | AU480 EIA Qualitative Result (mAU) | Pos/ Neg Result | Qualitative Cutoff Rate (mAU) |
|---|---|---|---|---|---|
| 37* | 1.5 | - | 85.9 | + | 83.0 |
| 41* | 2.7 | - | 111.3 | + | 83.0 |
| 43* | 3.0 | - | 207.9 | + | 83.0 |
| 44* | 3.0 | - | 107.7 | + | 83.0 |
| 45* | 3.3 | - | 124.7 | + | 83.0 |
| 46* | 3.5 | - | 169.6 | + | 83.0 |
| 47* | 3.8 | - | 204.6 | + | 83.0 |
| 48* | 3.9 | - | 113.6 | + | 83.0 |
| 49* | 4.2 | - | 263.1 | + | 83.0 |
- Values are discrepant below the cutoff concentration (0 ng/mL - 4.9 ng/mL)
These samples contained levels of fentanyl that contributed to the false positive result.
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Performance Characteristics Summary, continued: Beckman Coulter® AU480 Analyzer
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. Percent cross-reactivity was calculated by dividing the cutoff concentration by the lowest concentration of the compound where the assay response was near cutoff positive x 100.
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 | TestConcentration(ng/mL) | Qualitative Result(mAU) | QualitativeCutoff Rate(mAU) | % Cross-reactivity |
|---|---|---|---|---|
| Fentanyl | 3.8 | 107.8 | 84.6 | 131.58 % |
| Norfentanyl | 5 | 88.9 | 84.6 | 100.00 % |
Structurally Related Compounds:
| Compound | TestConcentration(ng/mL) | Qualitative Result(mAU) | QualitativeCutoff Rate(mAU) | % Cross-reactivity |
|---|---|---|---|---|
| 4-Fluoro-Isobutyryl Fentanyl | 20 | 84.5 | 83.3 | 25.00 % |
| 9-Hydroxy Risperidone | 100,000 | 1.8 | 84.6 | ND |
| Acetyl Fentanyl | 7 | 103.7 | 88.3 | 71.43 % |
| Acetyl Norfentanyl | 100 | 97.3 | 83.3 | 5.00 % |
| Acryl Fentanyl | 4 | 112.2 | 88.3 | 125.00 % |
| Alfentanil | 100,000 | 0.4 | 84.6 | ND |
| Butyryl Fentanyl | 6 | 99.7 | 88.3 | 83.33 % |
| Butyryl Norfentanyl | 40 | 96.4 | 83.3 | 12.50 % |
| Carfentanil Oxalate | 100,000 | 22.9 | 84.6 | ND |
| Cis- d,I 3-Methyl Fentanyl | 8 | 97.8 | 83.3 | 62.50 % |
| Cyclopropyl Fentanyl | 3.2 | 94.8 | 79.7 | 156.25 % |
| Cyclopropyl Norfentanyl | 25 | 89.7 | 83.9 | 20.00 % |
| Despropionyl Fentanyl (4-ANPP) | 100,000 | 0.0 | 88.3 | ND |
| Furanyl Fentanyl | 5.5 | 93.6 | 83.9 | 90.91 % |
| Furanyl Norfentanyl | 180 | 98.3 | 83.9 | 2.78 % |
| (±) ß-Hydroxy ThioFentanyl | 5 | 105.3 | 87.0 | 100.00 % |
| Isobutyryl Fentanyl | 15 | 87.5 | 83.9 | 33.33 % |
| Isobutyryl Norfentanyl | 500 | 81.3 | 79.5 | 1.00 % |
| Labetalol Hydrochloride | 100,000 | 0.0 | 84.6 | ND |
| Methoxyacetyl Fentanyl | 3.5 | 90.3 | 79.5 | 142.86 % |
| MT-45 | 100,000 | 0.0 | 84.6 | ND |
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Performance Characteristics Summary, continued:
Beckman Coulter® AU480 Analyzer
Beckman Coulter® AU480 Analyzer
Cross-reactivity, continued
Structurally Related Compounds, continued:
| Compound | TestConcentration(ng/mL) | Qualitative Result(mAU) | QualitativeCutoff Rate(mAU) | % Cross-reactivity |
|---|---|---|---|---|
| N-benzyl FuranylNorfentanyl | 11 | 82.1 | 79.5 | 45.45 % |
| N-benzyl Para-fluoroNorfentanyl | 4 | 89.1 | 79.5 | 125.00 % |
| Norcarfentanil Oxalate | 100,000 | 17.4 | 84.6 | ND |
| Ocfentanil | 3.8 | 116.7 | 79.5 | 131.58 % |
| Para-fluoro Butyryl Fentanyl(p-FBF) | 4.5 | 105.5 | 79.5 | 111.11 % |
| Para-fluoro Fentanyl | 3.2 | 122.1 | 79.5 | 156.25 % |
| Remifentanil | 100,000 | 0.2 | 84.6 | ND |
| Risperidone | 100,000 | 0.5 | 84.6 | ND |
| Sufentanil | 100,000 | 77.1 | 84.6 | ND |
| Thienyl Fentanyl | 4 | 89.7 | 79.5 | 125.00 % |
| Thiofentanyl | 3.2 | 92.6 | 79.7 | 156.25 % |
| Trans- d,I 3-Methyl Fentanyl | 6 | 96.3 | 79.7 | 83.33 % |
| Trazodone | 100,000 | 0.0 | 84.6 | ND |
| U-47700 | 100,000 | 0.0 | 84.6 | ND |
| Valeryl Fentanyl | 70 | 101.1 | 88.3 | 7.14 % |
| ω-1-Hydroxy Fentanyl | 300 | 94.8 | 79.7 | 1.67 % |
{11}------------------------------------------------
Beckman Coulter® AU480 Analyzer
Cross-reactivity, continued
Structurally unrelated compounds were additionally spiked into pooled negative human urine to described above). These solutions were then split into three portions; one without norfentany), and the remaining two that were further spiked with norfentanyl standards to a final norfentanyl concentration of 3.75 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). Samples were then evaluated against the assay's calbration curve in qualitative mode. All samples were tested in duplicates. Compounds tested at high concentration (100,000 ng/mL) with results below the cutoff value were listed as Not Detected (ND).
Interference was observed with Dextromethorphan at 40,000 ng/mL. No other significant cross-reactivity was observed.
| Compound | TestConcentration(ng/mL) | 0 ng/mLNorfentanyl% Cross | -25 % Norfentanyl Cutoff(3.75 ng/mL)Result | +25 % Norfentanyl Cutoff(6.25 ng/mL)Result |
|---|---|---|---|---|
| Acetaminophen | 100,000 | ND | Neg | Pos |
| 6-Acetylmorphine | 100,000 | ND | Neg | Pos |
| Acetylsalicylic Acid | 100,000 | ND | Neg | Pos |
| Amitriptyline | 100,000 | ND | Neg | Pos |
| Amlodipine Besylate | 100,000 | ND | Neg | Pos |
| Amoxicillin | 100,000 | ND | Neg | Pos |
| d-Amphetamine | 100,000 | ND | Neg | Pos |
| Atorvastatin | 100,000 | ND | Neg | Pos |
| Benzoylecgonine | 100,000 | ND | Neg | Pos |
| Buprenorphine | 100,000 | ND | Neg | Pos |
| Bupropion | 100,000 | ND | Neg | Pos |
| Caffeine | 100,000 | ND | Neg | Pos |
| Carbamazepine | 100,000 | ND | Neg | Pos |
| Cetirizine | 100,000 | ND | Neg | Pos |
| Compound | TestConcentration(ng/mL) | 0 ng/mLNorfentanyl% Cross | -25 % Norfentanyl Cutoff(3.75 ng/mL)Result | +25 % Norfentanyl Cutoff(6.25 ng/mL)Result |
| Chlorpheniramine | 100,000 | ND | Neg | Pos |
| Chlorpromazine | 100,000 | ND | Neg | Pos |
| Clomipramine | 100,000 | ND | Neg | Pos |
| Codeine | 100,000 | ND | Neg | Pos |
| Desipramine | 100,000 | ND | Neg | Pos |
| Dextromethorphan | 40,000 | 0.01 % | Pos | Pos |
| Diphenhydramine | 100,000 | ND | Neg | Pos |
| Duloxetine | 100,000 | ND | Neg | Pos |
| Fluoxetine | 100,000 | ND | Neg | Pos |
| Fluphenazine | 100,000 | ND | Neg | Pos |
| Gabapentin | 100,000 | ND | Neg | Pos |
| Hydrocodone | 100,000 | ND | Neg | Pos |
| Hydromorphone | 100,000 | ND | Neg | Pos |
| Ibuprofen | 100,000 | ND | Neg | Pos |
| Imipramine | 100,000 | ND | Neg | Pos |
| Lisinopril | 100,000 | ND | Neg | Pos |
| Losartan | 100,000 | ND | Neg | Pos |
| Loratidine | 100,000 | ND | Neg | Pos |
| MDA(3,4-methylene-dioxyamphetamine) | 100,000 | ND | Neg | Pos |
| MDEA | 100,000 | ND | Neg | Pos |
| MDMA(3,4-methylene-dioxymethamphetamine) | 100,000 | ND | Neg | Pos |
| Meperidine | 100,000 | ND | Neg | Pos |
| Compound | TestConcentration(ng/mL) | 0 ng/mLNorfentanyl% Cross | -25 % Norfentanyl Cutoff(3.75 ng/mL)Result | +25 % Norfentanyl Cutoff(6.25 ng/mL)Result |
| Metformin | 100,000 | ND | Neg | Pos |
| Metoprolol | 100,000 | ND | Neg | Pos |
| Methadone | 100,000 | ND | Neg | Pos |
| d-Methamphetamine | 100,000 | ND | Neg | Pos |
| Morphine | 100,000 | ND | Neg | Pos |
| Nalmefene | 100,000 | ND | Neg | Pos |
| Nicotine | 100,000 | ND | Neg | Pos |
| Nortriptyline | 100,000 | ND | Neg | Pos |
| Omeprazole | 100,000 | ND | Neg | Pos |
| Oxazepam | 100,000 | ND | Neg | Pos |
| Oxycodone | 100,000 | ND | Neg | Pos |
| Oxymorphone | 100,000 | ND | Neg | Pos |
| Phencyclidine | 100,000 | ND | Neg | Pos |
| Phenobarbital | 100,000 | ND | Neg | Pos |
| (1S,2S)-(+)Pseudoephedrine | 100,000 | ND | Neg | Pos |
| Quetiapine | 100,000 | ND | Neg | Pos |
| Ranitidine | 100,000 | ND | Neg | Pos |
| Salbutamol (Albuterol) | 100,000 | ND | Neg | Pos |
| Sertraline | 100,000 | ND | Neg | Pos |
| THC-COOH(11-Nor-Delta-9-THC-9-carboxylic acid) | 100,000 | ND | Neg | Pos |
| l-Thyroxine | 100,000 | ND | Neg | Pos |
| Tramadol | 100,000 | ND | Neg | Pos |
| Zolpidem | 100,000 | ND | Neg | Pos |
Structurally Unrelated Pharmacological Compounds:
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Beckman Coulter® AU480 Analyzer
Cross-reactivity, continued
Structurally Unrelated Pharmacological Compounds, continued:
{13}------------------------------------------------
Beckman Coulter® AU480 Analyzer
Cross-reactivity, continued
Structurally Unrelated Pharmacological Compounds, continued:
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
{14}------------------------------------------------
Beckman Coulter® AU480 Analyzer
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 norfentary), and the remaining two that were further spiked with norfentary) standards to a final norfentanyl concentration of 3.75 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). Samples were then evaluated against the assay's calbration curve in qualitative mode. All samples were tested in duplicates.
| Interfering Substance | Concentrationof Compound(mg/dL) | 0 ng/mLNorfentanyl | -25% Norfentanyl Cutoff(3.75 ng/mL) | +25% Norfentanyl Cutoff(6.25 ng/mL) |
|---|---|---|---|---|
| Acetone | 1000 | Neg | Neg | Pos |
| Ascorbic Acid | 500 | Neg | Neg | Pos |
| Bilirubin | 2 | Neg | Neg | Pos |
| Biotin | 0.5 | Neg | Neg | Pos |
| Boric Acid | 1000 | Neg | Neg | Neg |
| Calcium Chloride (CaCl2) | 300 | Neg | Neg | Pos |
| Citric Acid (pH 3) | 200 | Neg | Neg | Pos |
| Creatinine | 500 | Neg | Neg | Pos |
| Ethanol | 1000 | Neg | Neg | Pos |
| Galactose | 10 | Neg | Neg | Pos |
| y-Globulin | 500 | Neg | Neg | Pos |
| Glucose | 3000 | Neg | Neg | Pos |
| Hemoglobin | 300 | Neg | Neg | Pos |
| HSA | 500 | Neg | Neg | Pos |
| Human Urine (pooled) | N/A | Neg | Neg | Pos |
| ß-hydroxybutyric Acid | 100 | Neg | Neg | Pos |
| Oxalic Acid | 100 | Neg | Neg | Pos |
| Potassium Chloride | 1000 | Neg | Neg | Pos |
| Riboflavin | 7.5 | Neg | Neg | Pos |
| Sodium Azide | 1000 | Neg | Neg | Pos |
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Beckman Coulter® AU480 Analyzer
| Interfering Substance | Concentration ofCompound(mg/dL) | 0 ng/mLNorfentanyl | -25 % Norfentanyl Cutoff(3.75 ng/mL) | +25 % Norfentanyl Cutoff(6.25 ng/mL) |
|---|---|---|---|---|
| Sodium Chloride | 1000 | Neg | Neg | Pos |
| Sodium Fluoride | 1000 | Neg | Neg | Pos |
| Sodium Phosphate | 300 | Neg | Neg | Pos |
| Urea | 6000 | Neg | Neg | Pos |
| Uric Acid | 10 | Neg | Neg | Pos |
| LZI Urine-base Calibrator Buffer | N/A | Neg | Neg | Pos |
Endogenous and Preservative Compound Interference, continued:
The following endogenous and preservative compounds which showed interference at ±25 % of cutoff concentrations were then spiked into negative urine and at ±50 % of the cutoff concentration (2.5 ng/mL ) for the assay.
| Interfering Substance | Concentration of Compound (mg/dL) | 0 ng/mL Norfentanyl | -50 % Norfentanyl Cutoff (2.5 ng/mL) | +50 % Norfentanyl Cutoff (7.5 ng/mL) |
|---|---|---|---|---|
| Boric Acid | 1000 | Neg | Neg | Neg |
Interference was observed with Boric Acid at 1 % w/v. No other significant cross-reactants or endogenous/preservative substance interference was observed.
{16}------------------------------------------------
Beckman Coulter® AU480 Analyzer
Specific Gravity Interference:
Samples ranging in specific gravity from 1.000 to 1.027 were split into three portions each and either left un-spiked or further spiked to a final norfentanyl concentration of either 3.75 ng/mL or 6.25 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). These samples were then evaluated in qualitative mode. No interference was observed.
| SpecificGravityValue | 0 ng/mLNorfentanyl | -25 % Norfentanyl Cutoff(3.75 ng/mL) | +25 % Norfentanyl Cutoff(6.25 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 |
| 1.012 | Neg | Neg | Pos |
| 1.015 | Neg | Neg | Pos |
| 1.018 | Neg | Neg | Pos |
| 1.020 | Neg | Neg | Pos |
| 1.022 | Neg | Neg | Pos |
| 1.025 | Neg | Neg | Pos |
| 1.027 | Neg | Neg | Pos |
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Performance Characteristics Summary, continued: Beckman Coulter® AU480 Analyzer
pH Interference:
Negative urine and urine spiked with norfentanyl to the final norfentanyl concentration of either 3.75 ng/mL or 6.25 ng/mL (as negative or positive controls, ±25 % 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:
| InterferingSubstance | 0 ng/mLNorfentanyl | -25 % Norfentanyl Cutoff(3.75 ng/mL) | +25 % Norfentanyl Cutoff(6.25 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 |
Summary:
The information provided in this pre-market notification demonstrates that the LZI Fentanyl II 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 II 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).