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.

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.

AI/ML Overview

The provided text describes the LZI Cotinine II Enzyme Immunoassay, its intended use, and performance characteristics, but it does not describe a study that directly establishes acceptance criteria and directly proves the device meets specific acceptance criteria in a structured manner you requested.

The document is a 510(k) Summary of Safety and Effectiveness, which typically outlines the device's characteristics and demonstrates substantial equivalence to a predicate device. It includes performance data such as precision, linearity, and method comparison (accuracy) with clinical samples, which implicitly serve to show the device is safe and effective for its stated intended use. However, explicit acceptance criteria values that the device must meet are not directly listed in a table format with corresponding proof of meeting them.

I will extract the closest information available to address your request, making inferences where necessary based on the context of common performance expectations for such devices.

Here's a breakdown of the information as requested, largely derived from the "Method Comparison - Clinical Samples" and "Precision" sections.

Acceptance Criteria and Device Performance for LZI Cotinine II Enzyme Immunoassay

Note: The document does not explicitly state "acceptance criteria" in a definitive table format with corresponding proof. Instead, it presents performance study results that are implicitly considered acceptable for demonstrating substantial equivalence. The "Reported Device Performance" below is directly from the study results. The "Acceptance Criteria" are inferred based on the observed performance and typical expectations for diagnostic assays.

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Inferred Acceptance Criterion (Approximate)	Reported Device Performance (from Clinical Samples)	Study Comments
Accuracy (Semi-Quantitative)	High overall agreement with LC/MS, with minimal discrepancies, especially away from the cutoff concentration.	Overall Agreement: ~96.4% for positive, ~98.7% for negative. Discrepant Samples: - 1 sample (128.6 ng/mL LC/MS) was positive by EIA but was between 50% below cutoff and cutoff concentration (100-199.9 ng/mL). The report states it was "discrepant" but the EIA result (positive) aligns with the stated LC/MS value being within the range where a positive result starts to become more likely if it's "near cutoff positive". However, interpreting that specific discrepancy is complex without a clear cut-off definition of "positive" for LC/MS in the table. - 1 sample (204.2 ng/mL LC/MS) was negative by EIA but was between cutoff and 50% above cutoff concentration (200-299.9 ng/mL). The report states it was "discrepant".	The accuracy study uses LC/MS as the gold standard. The reported percentages reflect agreement for positive and negative results with respect to the 200 ng/mL cut-off. The discrepancies highlight inherent variability near the cutoff.
Accuracy (Qualitative)	High overall agreement with LC/MS, similar to semi-quantitative.	Overall Agreement: ~96.4% for positive, ~98.7% for negative. Discrepant Samples: - 1 sample (128.6 ng/mL LC/MS) was positive by EIA (146.6 mAU) with a qualitative cutoff rate of 126.4 mAU. This sample was LC/MS positive but between 100-199.9 ng/mL, the EIA result was positive. - 1 sample (204.2 ng/mL LC/MS) was negative by EIA (93.8 mAU) with a qualitative cutoff rate of 126.4 mAU. This sample was LC/MS positive (204.2 ng/mL) but the EIA result was negative.	Similar to semi-quantitative accuracy, with concordance defined by the qualitative response based on mAU compared to a cutoff rate.
Precision (Qualitative)	At 200 ng/mL cutoff, some variability is expected. For concentrations significantly above or below the cutoff, 100% agreement with expected result (positive/negative) is expected.	At 200 ng/mL: - Within Run (N=22): 10 Neg/12 Pos - Total Precision (N=88): 51 Neg/37 Pos At 250-400 ng/mL: 100% Positive At 0-150 ng/mL: 100% Negative	This indicates that at the exact cutoff concentration (200 ng/mL), there is expected variability in classification. This is typical for assays and is why results near the cutoff require careful interpretation and often confirmatory testing.
Precision (Semi-Quantitative)	Same as qualitative. At 200 ng/mL cutoff, some variability is expected. For concentrations significantly above or below the cutoff, 100% agreement with expected result (positive/negative) is expected.	At 200 ng/mL: - Within Run (N=22): 14 Neg/8 Pos - Total Precision (N=88): 55 Neg/33 Pos At 250-400 ng/mL: 100% Positive At 0-150 ng/mL: 100% Negative	The semi-quantitative precision results also show variability at the 200 ng/mL cutoff, with 100% agreement for samples sufficiently above or below the cutoff. This confirms the device's ability to consistently classify samples away from the decision point.
Linearity	Percent recovery between 85% and 115% of expected values across the assay's linear range.	Range 100 ng/mL to 1000 ng/mL: Recovery ranging from 97.9% to 110.8%. At 20 ng/mL: 167.5% recovery. At 0 ng/mL: N/A (reported 19.3 ng/mL determined).	The device demonstrated good linearity within the critical range for its intended use (100-1000 ng/mL). The higher recovery at 20 ng/mL suggests reduced accuracy at very low concentrations, which is generally not a concern given the 200 ng/mL cutoff.
Cross-reactivity	Minimal or no significant cross-reactivity with common substances, particularly structurally unrelated compounds, at physiologically relevant concentrations or concentrations higher than expected.	Cotinine-related compounds: Some cross-reactivity with compounds like (-) Norcotinine (20.00%) and (R, S)-Norcotinine (23.53%). Structurally Unrelated Pharmacological Compounds: "ND" (Not Detected) at high concentrations (e.g., 100,000 ng/mL) for most tested drugs. Negative results for 0 ng/mL cotinine spiked with these compounds, and correct positive/negative results for cotinine controls.	The device largely demonstrates specificity against a wide panel of common drugs and related substances, indicating minimal interference for its intended purpose. Some expected cross-reactivity with specific cotinine metabolites is noted.
Endogenous Interference	No significant interference from common endogenous substances at relevant concentrations.	No significant undesired cross-reactants or endogenous substance interference was observed except for Boric Acid (1000 ng/mL) and Citric Acid (pH 3) (800 ng/mL) which showed interference at ±25% of cutoff and also at ±50% of cutoff.	Most endogenous substances showed no interference. Boric acid and citric acid showed interference, which is important for laboratories to be aware of when testing samples that might contain these substances.
Specific Gravity Interference	No interference observed across the tested range.	No interference was observed in samples ranging from 1.005 to 1.028 specific gravity when spiked with cotinine at control concentrations.	This indicates the device is robust to variations in urine specific gravity, which is a common factor in urine drug testing.
pH Interference	No major interference observed across the tested pH range.	No major interference between pH 3 to pH 11.	This demonstrates the device's robustness across a wide physiological pH range for urine, an important aspect for clinical utility.

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

Accuracy (Method Comparison - Clinical Samples):
- Sample Size: 104 unaltered clinical samples.
- Data Provenance: Samples were collected by Lin-Zhi International, Inc. (LZI). The country of origin is not specified, but LZI is based in Santa Clara, CA, USA. The samples are indicated to be "clinical samples," implying they were prospectively collected from human subjects for diagnostic purposes. They are referred to as "unaltered," suggesting they were tested as received without intentional modification. The study is retrospective in the sense that these samples were then tested against a new assay.
Precision:
- Sample Size: For each cotinine concentration, 22 determinations were made within a run, and a total of 88 determinations were made across multiple runs (2 runs/day for 22 days).
- Data Provenance: Samples were prepared by spiking a cotinine standard into a "pool of negative human urine." This indicates a controlled laboratory setting (likely in-house at LZI) rather than clinical samples, and thus not tied to a specific country of origin from patients.
Linearity, Cross-reactivity, Endogenous Interference, Specific Gravity, pH Interference: These studies used spiked samples prepared in a laboratory setting (e.g., "drug free-urine pool," "pool of negative human urine"). The sample sizes for each condition are detailed in the tables (e.g., "10 replicates" for linearity, "replicates" for others).

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

For the Accuracy (Method Comparison - Clinical Samples): The ground truth was established by LC/MS (Liquid Chromatography/Mass Spectrometry). This is an analytical laboratory method, not typically performed by "experts" in the sense of clinicians or radiologists. The result from the LC/MS machine is the ground truth. Therefore, the concept of "number of experts" and their "qualifications" is not applicable here as the ground truth is an objective chemical measurement.

4. Adjudication Method for the Test Set

For the accuracy study, the "ground truth" was established by LC/MS, which is a definitive analytical method. Therefore, no human adjudication method (like 2+1, 3+1 consensus) was used or required for determining the confirmed cotinine concentration. The comparison was directly between the LZI Cotinine II Enzyme Immunoassay results and the LC/MS results.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This type of study is typical for imaging devices where human readers interpret images, sometimes with and without AI assistance, to assess AI's impact on human performance. The LZI Cotinine II Enzyme Immunoassay is an in-vitro diagnostic (IVD) device (a laboratory test), not an imaging device that would involve human readers for interpretation in this manner.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Yes, the performance studies described are essentially standalone evaluations of the LZI Cotinine II Enzyme Immunoassay. The accuracy, precision, linearity, and interference studies assess the analytical performance of the device itself (the "algorithm" in a broad sense, referring to the assay's methodology and reagents) against established reference methods or known concentrations, without requiring human intervention for interpretation beyond operating the automated clinical analyzer and analyzing the data. The results (e.g., ng/mL concentrations or positive/negative classifications) are generated directly by the assay on the automated analyzer.

7. The Type of Ground Truth Used

Primary Ground Truth for Clinical Accuracy: LC/MS (Liquid Chromatography/Mass Spectrometry) for cotinine concentrations. This is a highly specific and sensitive analytical technique, often considered the gold standard for drug confirmation testing.
Ground Truth for Other Studies (Precision, Linearity, Cross-reactivity, etc.): Known concentrations of cotinine or other substances (spiked samples) in negative human urine pools.

8. The Sample Size for the Training Set

The document describes performance studies for device validation. It does not provide information about a "training set" in the context of an AI/machine learning model. This device is an enzyme immunoassay, a chemical assay, not an AI-based diagnostic algorithm that typically undergoes a separate training phase with a large dataset. The "training" for such an assay would be its initial development and optimization, not a dataset-driven training process in the AI sense.

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

As explained in point 8, the concept of a "training set" for an AI/machine learning model is not applicable to this enzyme immunoassay device. Therefore, how its ground truth was established is not relevant here. The development of the assay's chemical reagents and methodology would have involved internal validation and optimization, but not a "training set" in the context of AI.

Summary

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left 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)

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 Druq 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."

{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 MetabolitesClass 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 kit comprised of two reagents, an R1 and R2 which are bottled separately but sold together within the kit.

{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.

DeviceCharacteristics	Subject Device (K192299)LZI Cotinine II Enzyme Immunoassay	Predicate Device (K963733)NicCheckI Test Strips
Intended Use	The LZI Cotinine II Enzyme Immunoassay isintended for the qualitative and semi-quantitative determination of cotinine inhuman urine at the cutoff value of200 ng/mL when calibrated against cotinine.The assay is intended as an aid in thedetection of cotinine after use or exposure totobacco products. The assay is designed forprescription use with a number of automatedclinical chemistry analyzers.The semi-quantitative mode is for purposesof (1) enabling laboratories to determine anappropriate dilution of the specimen forconfirmation by a confirmatory method suchas gas or liquid chromatography/massspectrometry (GC/MS or LC/MS) or (2)permitting laboratories to establish qualitycontrol procedures.This assay provides a rapid screening procedure fordetermining the presence of cotinine in urine. Theassay provides only a preliminary analytical result. Amore specific alternative chemical method must beused in order to obtain a confirmed analytical result.Gas or liquid chromatography/mass spectrometry(GC/MS or LC/MS) is the preferred confirmatorymethod. Clinical consideration and professionaljudgment should be exercised with any drug of abusetest result, particularly when the preliminary test resultis positive.	NicCheck I Test Strips may be used to detectnicotine and/or its metabolites in urine as anaid in indicating the smoking status of theindividual and in planning appropriatetreatment. The test will also aid in theidentification of a smoker as a low or highnicotine consumer.The NicCheck I test is for professional useonly. Individuals, such as physicians, nurses,pharmacists, medical technologists, andparamedical 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 ofmanufacture. Test strips aresusceptible to conditions of highhumidity, the canister must be kepttightly closed after removal of therequired 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.

CotinineConcentration	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)
CotinineConcentration	% of Cutoff	# ofDetermination	EIAResult	# ofDetermination	EIAResult
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)
CotinineConcentration	% of Cutoff	# ofDetermination	EIAResult	# ofDetermination	EIAResult
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.

CandidateDeviceResults	Negative	<50 %ofCutoff	Near CutoffNegative(between -50% of cutoffto the cutoff)	Near CutoffPositive(between cutoffand +50 % ofcutoff)	HighPositive(>50 %abovecutoff)	%Agreement
Positive	0	0	1*	11	16	96.4 %
Negative	20	32	23	1**	0	98.7 %

Sample#	LC/MSMethadone(ng/mL)	Pos/NegResult	AU480 EIAPos/NegResult
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.

CandidateDeviceResults	Negative	<50 %ofCutoff	Near CutoffNegative(between -50% of cutoffto the cutoff)	Near CutoffPositive(between cutoffand +50 % ofcutoff)	HighPositive(>50 %abovecutoff)	%Agreement
Positive	0	0	1*	11	16	96.4 %
Negative	20	32	23	1**	0	98.7 %

Sample#	LC/MSMethadone(ng/mL)	Pos/NegResult	AU680 EIAQualitativeResult (mAU)	Pos/NegResult	QualitativeCutoff 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	TargetConcentration(ng/mL)	% Cross-reactivity
(-)-Cotinine	200	100.00%

Cotinine Metabolites:

Compound	TargetConcentration(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/mLControl	250 ng/mLControl
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/mLControl	250 ng/mLControl
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	ConcentrationTested (ng/mL)	Spiked Cotinine Concentration
Boric Acid	1000	0 ng/mLNeg	100 ng/mLControlNeg	300 ng/mLControlNeg
Citric Acid (pH 3)	800	0 ng/mLNeg	100 ng/mLControlNeg	300 ng/mLControlNeg

{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/mLControl	250 ng/mLControl
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.

Regulation Number and Section

§ 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.