(90 days)
Beckman Coulter® AU480 automated clinical analyzer
No
The device description details a standard enzyme immunoassay based on chemical reactions and spectrophotometric measurement. There is no mention of AI or ML in the intended use, device description, performance studies, or any other section of the summary. The analysis is based on direct chemical interaction and measurement of enzyme activity.
No
Explanation: This device is for in vitro diagnostic use, specifically for the quantitative determination of cotinine in human urine as an aid in detecting tobacco use/exposure. It does not directly provide therapy or treatment to a patient.
Yes
Explanation: The device is intended "for the quantitative determination of cotinine in human urine" and "as an aid in the detection of cotinine after use or exposure to tobacco products." This directly states its purpose in providing information for a medical diagnosis or condition.
No
The device is a homogeneous enzyme immunoassay kit comprised of liquid reagents (R1 and R2) used with automated clinical chemistry analyzers, indicating it is a hardware-based diagnostic test, not software only.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states "quantitative determination of cotinine in human urine" and "aid in the detection of cotinine after use or exposure to tobacco products." This clearly indicates the device is used to test a sample taken from the human body (urine) to provide information about a physiological state (exposure to tobacco products).
- Device Description: The description details a "homogeneous enzyme immunoassay" that measures enzyme activity in a sample. This is a common method used in in vitro diagnostic tests.
- Performance Studies: The document describes performance studies conducted on "unaltered clinical samples" (human urine) to evaluate the assay's precision, linearity, and accuracy against a confirmatory method (LC/MS). This type of testing is characteristic of IVD validation.
- Regulatory Context: The mention of "prescription use" and being designed for use with "automated clinical chemistry analyzers" further supports its classification as a medical device intended for use in a clinical laboratory setting, which is typical for IVDs.
- Predicate Device: The inclusion of a predicate device (K963733; NicCheckI Test Strips) which is also an IVD, suggests that this device is being compared to a similar product already on the market and regulated as an IVD.
All these factors align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological state, state of health, or disease.
N/A
Intended Use / Indications for Use
The LZI Cotinine II Enzyme Immunoassay is intended for the quantitative determination of cotinine in human urine at the cutoff value of 200 ng/mL when calibrated against cotinine. The assay is intended as an aid in the detection of cotinine after use or exposure to tobacco products. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas or liquid chromatography/mass spectrometry (GC/MS or LC/MS) or (2) permitting laboratories to establish quality control procedures.
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.
Product codes
MKU
Device Description
The LZI Cotinine 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. 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, cotinine-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 cotinine-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 Cotinine 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 R1 solution contains mouse monoclonal anti-cotinine 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 cotinine in buffer with sodium azide (0.09 %) as a preservative.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
prescription use with a number of automated clinical chemistry analyzers.
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies
Precision: The assay was tested in qualitative (ΔOD, mAU) and semi-quantitative (ng/mL) mode using a modified NCCLS-EP5 protocol. Cotinine sample concentrations were prepared by spiking a cotinine standard into a pool of negative human urine at concentrations ±25%, ±50%, ±75%, and ±100% of cutoff concentration. Results were obtained by testing all samples in replicate of two, two runs a day for 22 days on one AU480 automatic clinical analyzer for a total of 88 runs.
Linearity: A drug free-urine pool spiked with cotinine at 1000 ng/mL was serially diluted. Each sample was run in 10 replicates and the average was used to determine percent recovery compared to the expected target value. Recovery values (Expected Value divided by the Observed Value) were considered acceptable between 85 — 115%. Samples from the linear range of the assay (100 ng/mL to 1000 ng/mL) were tested with recovery ranging from 97.9% to 110.8%.
Method Comparison - Clinical Samples: A total of one-hundred and four (104) unaltered clinical samples were tested with the LZI Cotinine II Enzyme Immunoassay on the Beckman Coulter® AU480 automated clinical analyzer. Samples were evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. All samples were tested in singlet. All samples were confirmed with LC/MS for cotinine concentrations. Semi-Quantitative Accuracy Study showed 96.4% agreement for positive results and 98.7% agreement for negative results. Qualitative Accuracy Study also showed 96.4% agreement for positive results and 98.7% agreement for negative results.
Cross-reactivity: The Cross-reactivity of various potentially interfering drugs and endogenous compounds were tested by spiking various concentrations into a pool of negative human urine and then evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes.
Specific Gravity: Samples ranging in specific gravity from 1.005 to 1.028 were split into three portions each and either left un-spiked or further spiked to a final methadone concentration of either 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively). These samples were then evaluated in qualitative mode. No interference was observed.
pH Interference Study: Negative urine and urine spiked with cotinine to the final cotinine concentration of either 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively) were adjusted to various pH levels (3 to 11) and tested by the assay. No major interference between pH 3 to pH 11 was observed.
Key Metrics
Semi-Quantitative Accuracy Study:
Positive Agreement: 96.4 %
Negative Agreement: 98.7 %
Qualitative Accuracy Study:
Positive Agreement: 96.4 %
Negative Agreement: 98.7 %
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 862.3220 Carbon monoxide test system.
(a)
Identification. A carbon monoxide test system is a device intended to measure carbon monoxide or carboxyhemoglobin (carbon monoxide bound to the hemoglobin in the blood) in blood. Measurements obtained by this device are used in the diagnosis and treatment of or confirmation of carbon monoxide poisoning.(b)
Classification. Class I (general controls). 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.
0
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 and 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.
November 21, 2019
Lin-Zhi International, Inc. Bernice Lin VP of Operations 2945 Oakmead Village Court Santa Clara, CA 95051
Re: K192299
Trade/Device Name: LZI Cotinine II Enzyme Immunoassay Regulation Number: 21 CFR 862.3220 Regulation Name: Carbon monoxide test system Regulatory Class: Class I, reserved Product Code: MKU Dated: August 19, 2019 Received: August 23, 2019
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.
1
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 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 -S
Marianela Perez-Torres, Ph.D. Deputy Director (Acting) 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
2
Indications for Use
510(k) Number (if known) K192299
Device Name
LZI Cotinine II Enzyme Immunoassay
Indications for Use (Describe)
The LZI Cotinine II Enzyme Immunoassay is intended for the quantitative determination of cotinine in human urine at the cutoff value of 200 ng/mL when calibrated against cotinine. The assay is intended as an aid in the detection of cotinine after use or exposure to tobacco products. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas or liquid chromatography/mass spectrometry (GC/MS or LC/MS) or (2) permitting laboratories to establish quality control procedures.
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) |
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3
510(k) Number: K192299
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.
Submitted On
August 21, 2019
Last Updated On
November 20, 2019
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
Bernice Lin, Ph.D. Contact: VP Operations
Device Name and Classification
| Classification Name: | Enzyme Immunoassay, Nicotine and Nicotine Metabolites
Class II, MKU (91 Toxicology),
21 CFR 862.3220 |
|----------------------|------------------------------------------------------------------------------------------------------------|
| Common Name: | Homogeneous Cotinine Enzyme Immunoassay |
| Proprietary Name: | LZI Cotinine II Enzyme Immunoassay |
4
Legally Marketed Predicate Device(s)
The LZI Cotinine II Enzyme Immunoassay is substantially equivalent to the NicCheckI Test Strips (K963733) manufactured by Dynagen, Inc. The LZI Cotinine 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 Cotinine 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. 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, cotinine-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 cotinine-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 Cotinine 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 R1 solution contains mouse monoclonal anti-cotinine 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 cotinine in buffer with sodium azide (0.09 %) as a preservative.
5
Intended Use
The LZI Cotinine II Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of cotinine in human urine at the cutoff value of 200 ng/mL when calibrated against cotinine. The assay is intended as an aid in the detection of cotinine after use or exposure to tobacco products. The assay is designed for prescription use with a number of automated clinical chemistry analyzers.
The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas or liquid chromatography/mass spectrometry (GC/MS or (2) permitting laboratories to establish quality control procedures.
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.
6
Comparison to Predicate Device
The LZI Cotinine II Enzyme Immunoassay is substantially equivalent to the NicCheckI Test Strips manufactured by Dynagen, Inc. which was cleared by the FDA under the premarket notification K963733 for its stated intended use.
The following table compares the LZI Cotinine II Enzyme Immunoassay with the predicate device.
| Device
Characteristics | Subject Device (K192299)
LZI Cotinine II Enzyme Immunoassay | Predicate Device (K963733)
NicCheckI Test Strips |
|---------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The LZI Cotinine II Enzyme Immunoassay is
intended for the qualitative and semi-
quantitative determination of cotinine in
human urine at the cutoff value of
200 ng/mL when calibrated against cotinine.
The assay is intended as an aid in the
detection of cotinine after use or exposure to
tobacco products. The assay is designed for
prescription use with a number of automated
clinical chemistry analyzers.
The semi-quantitative mode is for purposes
of (1) enabling laboratories to determine an
appropriate dilution of the specimen for
confirmation by a confirmatory method suchas gas or liquid chromatography/mass
spectrometry (GC/MS or LC/MS) or (2)
permitting laboratories to establish quality
control procedures.
This assay provides a rapid screening procedure for
determining the presence of cotinine 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. | NicCheck I Test Strips may be used to detect
nicotine and/or its metabolites in urine as an
aid in indicating the smoking status of the
individual and in planning appropriate
treatment. The test will also aid in the
identification of a smoker as a low or high
nicotine consumer.
The NicCheck I test is for professional use
only. Individuals, such as physicians, nurses,
pharmacists, medical technologists, and
paramedical personnel may perform the test. |
| Analyte | cotinine | cotinine |
| Cutoff | 200 ng/mL | 200 ng/mL |
| Matrix | urine | urine |
| Calibrators Level | 0, 100, 200, 400, and 1000 ng/mL | N/A |
| Controls Level | 150 ng/mL and 250 ng/mL | N/A |
| Storage | 2-8 oC until expiration date. | 2-8 oC until 2 years from date of
manufacture. Test strips are
susceptible to conditions of high
humidity, the canister must be kept
tightly closed after removal of the
required number of strips. |
7
Precision: 200 ng/mL Cutoff
The assay was tested in qualitative (ΔΟD, mAU) and semi-quantitative (ng/mL) mode using a modified NCCLS-EP5 protocol. Cotinine sample concentrations were prepared by spiking a cotinine standard into a pool of negative human urine at concentrations ±25%, ±50%, ±75%, and ±100% of 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 AU480 automatic clinical analyzer for a total of 88 runs. Samples were evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. One single lot of reagents, calibrators, and controls were used and stored at 2-8°C when not in use.
| Cotinine
Concentration | Within Run (N=22)
Qualitative Response | Total Precision (N=88)
Qualitative Response |
|---------------------------|-------------------------------------------|------------------------------------------------|
| 0 ng/mL | - | - |
| 50 ng/mL | - | - |
| 100 ng/mL | - | - |
| 150 ng/mL | - | - |
| 200 ng/mL | - | - |
| 250 ng/mL | + | + |
| 300 ng/mL | + | + |
| 350 ng/mL | + | + |
| 400 ng/mL | + | + |
Semi-Quantitative Precision Analysis Summary: Qualitative Results
| 200 ng/mL Cutoff Result: | Within Run (N=22) | Total Precision (N=88) | |||
|---|---|---|---|---|---|
| Cotinine | |||||
| Concentration | % of Cutoff | # of | |||
| Determination | EIA | ||||
| Result | # of | ||||
| Determination | EIA | ||||
| Result | |||||
| 0 ng/mL | 0.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 50 ng/mL | 25.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 100 ng/mL | 50.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 150 ng/mL | 75.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 200 ng/mL | 100.0 % | 22 | 14 Neg/8 Pos | 88 | 55 Neg/33 Pos |
| 250 ng/mL | 125.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 300 ng/mL | 150.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 350 ng/mL | 175.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 400 ng/mL | 200.0 % | 22 | 22 Positive | 88 | 88 Positive |
Semi-Ouantitative Positive/Negative Results:
8
| 200 ng/mL Cutoff Result: | Within Run (N=22) | Total Precision (N=88) | |||
|---|---|---|---|---|---|
| Cotinine | |||||
| Concentration | % of Cutoff | # of | |||
| Determination | EIA | ||||
| Result | # of | ||||
| Determination | EIA | ||||
| Result | |||||
| 0 ng/mL | 0.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 50 ng/mL | 25.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 100 ng/mL | 50.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 150 ng/mL | 75.0 % | 22 | 22 Negative | 88 | 88 Negative |
| 200 ng/mL | 100.0 % | 22 | 10 Neg/12 Pos | 88 | 51 Neg/37 Pos |
| 250 ng/mL | 125.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 300 ng/mL | 150.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 350 ng/mL | 175.0 % | 22 | 22 Positive | 88 | 88 Positive |
| 400 ng/mL | 200.0 % | 22 | 22 Positive | 88 | 88 Positive |
Qualitative Positive/Negative Results:
Linearity:
To demonstrate linearity of the entire assay range, a drug free-urine pool spiked with cotinine at 1000 ng/mL was serially diluted. Each sample was run in 10 replicates and the average was used to determine percent recovery compared to the expected target value.
Observed values were obtained and acceptable if measurements were ±15% of the expected values. Recovery values (Expected Value divided by the Observed Value) were considered acceptable between 85 — 115%.
Samples from the linear range of the assay (100 ng/mL to 1000 ng/mL) were tested with recovery ranging from 97.9% to 110.8%.
| Target Concentration
(ng/mL) | Determined
(ng/mL) | % Recovery |
|---------------------------------|-----------------------|------------|
| 1000 | 978.9 | 97.9% |
| 900 | 919.6 | 102.2% |
| 800 | 851.7 | 106.5% |
| 700 | 766.0 | 109.4% |
| 600 | 664.8 | 110.8% |
| 500 | 539.1 | 107.8% |
| 400 | 395.0 | 98.8% |
| 300 | 302.4 | 100.8% |
| 200 | 199.9 | 100.0% |
| 100 | 109.0 | 109.0% |
| 20 | 33.5 | 167.5% |
| 0 | 19.3 | N/A |
9
Method Comparison - Clinical Samples:
A total of one-hundred and four (104) unaltered clinical samples were tested with the LZI Cotinine II Enzyme Immunoassay on the Beckman Coulter® AU480 automated clinical analyzer. Samples were evaluated against the assay's calibration curve in both qualitative and semiquantitative modes. All samples were tested in singlet.
All samples were confirmed with LC/MS for cotinine concentrations. Samples were collected by Lin-Zhi International, Inc. (LZI).
Semi-Quantitative Accuracy Study:
Discrepant samples determined as compared to cotinine concentration from LC/MS.
| Candidate
Device
Results | Negative | 50 %
above
cutoff) | %
Agreement |
|--------------------------------|----------|-----------------------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|------------------------------------------------|----------------|
| Positive | 0 | 0 | 1* | 11 | 16 | 96.4 % |
| Negative | 20 | 32 | 23 | 1** | 0 | 98.7 % |
| Sample
| LC/MS
Methadone
(ng/mL) | Pos/Neg
Result | AU480 EIA
Pos/Neg
Result |
|-------------|-------------------------------|-------------------|--------------------------------|
| 57* | 128.6 | + | + |
| 78** | 204.2 | + | - |
- Discrepant between 50% below cutoff and cutoff concentration (100 - 199.9 ng/mL)
** Discrepant between cutoff and 50% above cutoff concentration (200 - 299.9 ng/mL)
Qualitative Accuracy Study:
Discrepant samples determined as compared to cotinine concentration from LC/MS.
| Candidate
Device
Results | Negative | 50 %
above
cutoff) | %
Agreement |
|--------------------------------|----------|-----------------------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|------------------------------------------------|----------------|
| Positive | 0 | 0 | 1* | 11 | 16 | 96.4 % |
| Negative | 20 | 32 | 23 | 1** | 0 | 98.7 % |
| Sample
| LC/MS
Methadone
(ng/mL) | Pos/Neg
Result | AU680 EIA
Qualitative
Result (mAU) | Pos/
Neg
Result | Qualitative
Cutoff Rate
(mAU) |
|-------------|-------------------------------|-------------------|------------------------------------------|-----------------------|-------------------------------------|
| 57* | 128.6 | + | 146.6 | + | 126.4 |
| 78** | 204.2 | + | 93.8 | - | 126.4 |
- Discrepant between 50% below cutoff and cutoff concentration (100 - 199.9 ng/mL)
** Discrepant between cutoff and 50% above cutoff concentration (200 - 299.9 ng/mL)
10
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 both qualitative and semi-quantitative modes. All samples were tested in replicates.
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 with results below the cutoff value were listed as Not Detected (ND).
Cotinine:
| Compound | Target
Concentration
(ng/mL) | % Cross-
reactivity |
|--------------|------------------------------------|------------------------|
| (-)-Cotinine | 200 | 100.00% |
Cotinine Metabolites:
| Compound | Target
Concentration
(ng/mL) | % Cross-
reactivity |
|--------------------------|------------------------------------|------------------------|
| (+)-Anabasine | 250,000 | 0.08% |
| S(-)-Nicotine | 25,000 | 0.80% |
| Nicotinic Acid (Niacin) | 100,000 | 0.20% |
| (-) Norcotinine | 1000 | 20.00% |
| (+) Norcotinine | 100,000 | 0.20% |
| (R, S)-Norcotinine | 850 | 23.53% |
| (±)-Nornicotine | 250,000 | 0.08% |
| trans-3'-hydroxycotinine | 10,000 | 2.00% |
Structurally unrelated compounds were additionally spiked into pooled negative human urine to desired concentrations (as described above). These solutions were then split into three portions; one without cotinine, and the remaining two that were further spiked with cotinine standards to a final cotinine concentration of 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively). Samples were then evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. All samples were tested in replicates.
| | Spiked []
(ng/mL) | Spiked Cotinine Concentration | | |
|--------------------------------------------------------|----------------------|-------------------------------|----------------------|----------------------|
| Cross-reactant | | 0 ng/mL | 150 ng/mL
Control | 250 ng/mL
Control |
| Acetaminophen | 100,000 | ND | Neg | Pos |
| 6-Acetylmorphine | 100,000 | ND | Neg | Pos |
| Acetylsalicylic Acid | 100,000 | ND | Neg | Pos |
| Cross-reactant | Spiked []
(ng/mL) | Spiked Cotinine Concentration | | |
| | | | 150 ng/mL | 250 ng/mL |
| | | 0 ng/mL | Control | Control |
| Amitriptyline | 100,000 | ND | Neg | Pos |
| Amlodipine Besylate | 100,000 | ND | Neg | Pos |
| Amoxicillin | 100,000 | ND | Neg | Pos |
| Azelastine | 100,000 | ND | Neg | Pos |
| d-Amphetamine | 100,000 | ND | Neg | Pos |
| Atorvastatin | 20,000 | ND | Neg | Pos |
| Benzoylecgonine | 100,000 | ND | Neg | Pos |
| Brompheniramine | 100,000 | ND | Neg | Pos |
| Buprenorphine | 15,000 | ND | Neg | Pos |
| Bupropion | 100,000 | ND | Neg | Pos |
| Caffeine | 100,000 | ND | Neg | Pos |
| Carbamazepine | 100,000 | ND | Neg | Pos |
| Carbinoaxmine | 100,000 | ND | Neg | Pos |
| Cetirizine | 100,000 | ND | Neg | Pos |
| Chlorpheniramine | 100,000 | ND | Neg | Pos |
| Chlorpromazine | 100,000 | ND | Neg | Pos |
| Clemastine | 100,000 | ND | Neg | Pos |
| Clomipramine | 100,000 | ND | Neg | Pos |
| Codeine | 100,000 | ND | Neg | Pos |
| Cyproheptadine | 100,000 | ND | Neg | Pos |
| Desipramine | 100,000 | ND | Neg | Pos |
| Desloratadine | 100,000 | ND | Neg | Pos |
| Dexchlorpheniramine | 100,000 | ND | Neg | Pos |
| Diphenhydramine | 100,000 | ND | Neg | Pos |
| Doxylamine | 100,000 | ND | Neg | Pos |
| Duloxetine | 100,000 | ND | Neg | Pos |
| Emedastine | 100,000 | ND | Neg | Pos |
| Fentanyl (citrate) | 10,000 | ND | Neg | Pos |
| Fexofenadine | 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 |
| Hydroxyzine | 100,000 | ND | Neg | Pos |
| Ibuprofen | 100,000 | ND | Neg | Pos |
| Imipramine | 100,000 | ND | Neg | Pos |
| Lisinopril | 100,000 | ND | Neg | Pos |
| Levocabastine | 100,000 | ND | Neg | Pos |
| Losartan | 10,000 | ND | Neg | Pos |
| Cross-reactant | Spiked []
(ng/mL) | Spiked Cotinine Concentration | | |
| | | 0 ng/mL | 150 ng/mL
Control | 250 ng/mL
Control |
| Loratidine | 100,000 | ND | Neg | Pos |
| MDA (3,4-
methylenedioxyamphetamine) | 100,000 | ND | Neg | Pos |
| MDEA | 100,000 | ND | Neg | Pos |
| MDMA (3,4-
methylenedioxymethamphetamine) | 100,000 | ND | Neg | Pos |
| Meperidine | 100,000 | ND | Neg | Pos |
| Metformin | 100,000 | ND | Neg | Pos |
| Methapyrilene | 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 |
| 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 |
| Phenobarbital | 100,000 | ND | Neg | Pos |
| Promethazine | 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 |
| Terfenadine | 100,000 | ND | Neg | Pos |
| THC-COOH
(11-Nor-Delta-9-THC-9-
carboxylic acid) | 1,000 | ND | Neg | Pos |
| l-Thyroxine | 10,000 | ND | Neg | Pos |
| Tramadol | 100,000 | ND | Neg | Pos |
| Triprolidine | 100,000 | ND | Neg | Pos |
| Zolpidem | 10,000 | ND | Neg | Pos |
Structurally Unrelated Pharmacological Compounds:
11
Structurally Unrelated Pharmacological Compounds, continued:
12
Structurally Unrelated Pharmacological Compounds, continued:
13
Endogenous compounds were spiked into pooled negative human urine to desired concentrations. These solutions were then split into three portions; one without cotinine, and the remaining two that were further spiked with cotinine standards to a final cotinine concentration of 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively). Samples were then evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. All samples were tested in replicates.
| Endogenous Substance | Concentration Tested (ng/mL) | Spiked Cotinine Concentration | ||
|---|---|---|---|---|
| 0 ng/mL | 150 ng/mL Control | 250 ng/mL Control | ||
| Acetone | 1000 | Neg | Neg | Pos |
| Ascorbic Acid | 1500 | Neg | Neg | Pos |
| Bilirubin | 2 | Neg | Neg | Pos |
| Boric Acid | 1000 | Neg | Neg | Neg |
| Calcium Chloride (CaCl2) | 300 | Neg | Neg | Pos |
| Citric Acid (pH 3) | 800 | Neg | Neg | Neg |
| Creatinine | 500 | Neg | Neg | Pos |
| Ethanol | 1000 | Neg | Neg | Pos |
| Galactose | 10 | Neg | Neg | Pos |
| γ-Globulin | 500 | Neg | Neg | Pos |
| Glucose | 3000 | Neg | Neg | Pos |
| Hemoglobin | 300 | Neg | Neg | Pos |
| β-hydroxybutyric Acid | 100 | Neg | Neg | Pos |
| HAS | 500 | Neg | Neg | Pos |
| Oxalic Acid | 100 | Neg | Neg | Pos |
| Potassium Chloride | 6000 | Neg | Neg | Pos |
| Riboflavin | 0.3 | Neg | Neg | Pos |
| Urea | 6000 | Neg | Neg | Pos |
| Uric Acid | 10 | Neg | Neg | Pos |
| Sodium Azide | 1000 | Neg | Neg | Pos |
| Sodium Chloride | 6000 | Neg | Neg | Pos |
| Sodium Fluoride | 1000 | Neg | Neg | Pos |
| Sodium Phosphate | 300 | Neg | Neg | Pos |
The following endogenous compounds which showed interference at ±25 % of cutoff concentrations were then spiked into negative urine and at ±50 % of cutoff concentrations (100 ng/mL and 300 ng/mL) for the assay.
Interference was observed with Boric Acid at 1 % w/v and Citric Acid. No other significant undesired cross-reactants or endogenous substance interference was observed.
| Endogenous Substance | Concentration
Tested (ng/mL) | Spiked Cotinine Concentration | | |
|----------------------|---------------------------------|-------------------------------|-----------------------------|-----------------------------|
| Boric Acid | 1000 | 0 ng/mL
Neg | 100 ng/mL
Control
Neg | 300 ng/mL
Control
Neg |
| Citric Acid (pH 3) | 800 | 0 ng/mL
Neg | 100 ng/mL
Control
Neg | 300 ng/mL
Control
Neg |
14
Specific Gravity:
Samples ranging in specific gravity from 1.005 to 1.028 were split into three portions each and either left un-spiked or further spiked to a final methadone concentration of either 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively). These samples were then evaluated in qualitative mode. No interference was observed.
pH Interference Study:
Negative urine and urine spiked with cotinine to the final cotinine concentration of either 150 ng/mL or 250 ng/mL (as negative or positive controls, ±25% cutoff concentration, respectively) were adjusted to the following pH levels and tested by the assay. The pH adjusted solutions were evaluated against the cutoff calibrator.
| pH | Spiked Cotinine Concentration | ||
|---|---|---|---|
| 0 ng/mL | 150 ng/mL | ||
| Control | 250 ng/mL | ||
| Control | |||
| 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 |
No major interference between pH 3 to pH 11. Results are summarized in the following table:
Summary:
The information provided in this pre-market notification demonstrates that the LZI Cotinine 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 Cotinine II Enzyme Immunoassay is safe and effective for its stated intended use.