Who is the manufacturer of LZI Oxycodone III Enzyme Immunoassay?

Lin-Zhi International, Inc. filed the 510(k) submission for LZI Oxycodone III Enzyme Immunoassay (K202007).

What is the FDA product code for LZI Oxycodone III Enzyme Immunoassay?

DJG. It is classified under 21 CFR 862.3650 as a Class 2 device in the Clinical Toxicology panel.

What is the regulatory pathway for LZI Oxycodone III Enzyme Immunoassay?

510(k) clearance (submission type: Traditional).

What are the predicate devices for LZI Oxycodone III Enzyme Immunoassay?

LZI Oxycodone Homogeneous Enzyme Immunoassay, LZI Oxycondone Calibrators, LZI Oxycodone Controls (K120763).

Who can help prepare an FDA 510(k) submission like LZI Oxycodone III Enzyme Immunoassay?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

LZI Oxycodone III Enzyme Immunoassay

K202007 · Lin-Zhi International, Inc. · DJG · Sep 17, 2020 · Clinical Toxicology

Device Facts

Record ID	K202007
Device Name	LZI Oxycodone III Enzyme Immunoassay
Applicant	Lin-Zhi International, Inc.
Product Code	DJG · Clinical Toxicology
Decision Date	Sep 17, 2020
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 862.3650
Device Class	Class 2

Intended Use

The LZI Oxycodone III Enzyme Immunoassay is intended for the qualitative and semiquantitative determination of oxycodone in human urine at the cutoff values of 100 ng/mL and 300 ng/mL when calibrated against oxycodone. 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 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 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.

Device Story

LZI Oxycodone III Enzyme Immunoassay is a homogeneous enzyme immunoassay kit; utilizes two liquid reagents (R1: anti-oxycodone antibody; R2: oxycodone-labeled G6PDH conjugate). Operates via competitive binding: drug in urine sample competes with drug-labeled G6PDH for fixed antibody sites. Enzyme activity is inhibited when bound to antibody; free drug in sample displaces conjugate, increasing enzyme activity. Active enzyme converts NAD to NADH, producing absorbance change measured spectrophotometrically at 340 nm. Used in clinical laboratories on automated chemistry analyzers (e.g., Beckman Coulter AU480) by trained technicians. Provides preliminary qualitative/semi-quantitative results to guide specimen dilution for confirmatory testing (GC/MS or LC/MS). Assists clinicians in identifying potential oxycodone presence; requires professional judgment for clinical decision-making.

Clinical Evidence

Bench testing only. Precision studies (N=88 runs) evaluated qualitative and semi-quantitative performance at 100 ng/mL and 300 ng/mL cutoffs. Method comparison against LC/MS using clinical urine samples (N=82 for 100 ng/mL; N=90 for 300 ng/mL) showed high agreement. Cross-reactivity and interference studies (endogenous, preservative, and pharmacological compounds) confirmed assay specificity. No clinical prospective/retrospective studies were required.

Technological Characteristics

Homogeneous enzyme immunoassay; liquid reagents (R1: mouse monoclonal anti-oxycodone antibody, G6P, NAD; R2: oxycodone-labeled G6PDH). Sensing principle: spectrophotometric measurement of NADH production at 340 nm. Connectivity: compatible with automated clinical chemistry analyzers. Storage: 2-8 °C.

Indications for Use

Indicated for the qualitative and semi-quantitative detection of oxycodone in human urine at 100 ng/mL and 300 ng/mL cutoffs. For prescription use in clinical laboratories to aid in identifying specimens for confirmatory testing. Not for diagnostic use; results are preliminary and require confirmation by GC/MS or LC/MS.

Regulatory Classification

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.

Special Controls

*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).

Predicate Devices

LZI Oxycodone Homogeneous Enzyme Immunoassay, LZI Oxycondone Calibrators, LZI Oxycodone Controls (K120763)

Related Devices

K131168 — IMMUNALYSIS OXYCODONE ENZYME IMMUNOASSAY · Immunalysis Corporation · Jan 17, 2014

Submission Summary (Full Text)

{0}------------------------------------------------ Image /page/0/Picture/0 description: The image shows the logo for the U.S. Food & Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services seal on the left and the FDA acronym followed by the full name of the agency on the right. The FDA part of the logo is in blue, with the acronym in a solid blue square and the agency name in a lighter blue. The text reads "FDA U.S. FOOD & DRUG ADMINISTRATION". September 17, 2020 Lin-Zhi International, Inc. Bernice Lin VP of Operations 2945 Oakmead Village Court Santa Clara, CA 95051 Re: K202007 Trade/Device Name: LZI Oxycodone III Enzyme Immunoassay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: Class II Product Code: DJG Dated: July 17, 2020 Received: July 21, 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 {1}------------------------------------------------ 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 (OS) 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 {2}------------------------------------------------ #### Indications for Use 510(k) Number (if known) k202007 Device Name LZI Oxycodone III Enzyme Immunoassay Indications for Use (Describe) The LZI Oxycodone III Enzyme Immunoassay is intended for the quantitative determination of oxycodone in human urine at the cutoff values of 100 ng/mL when calibrated against oxycodone. 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 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. Type of Use (Select one or both, as applicable) > Prescription Use (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 801 Subpart C) #### CONTINUE ON A SEPARATE PAGE IF NEEDED. This section applies only to requirements of the Paperwork Reduction Act of 1995. #### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {3}------------------------------------------------ # 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 k202007 ### Prepared On September 14, 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 Fax: (408) 970-9030 e-mail: bclin@lin-zhi.com | Contact: | Bernice Lin, Ph.D. | |----------|--------------------| | | VP of Operations | #### Device Name and Classification | Classification Name: | Enzyme Immunoassay, Oxycodone Class II, DJG (91 Toxicology) 21 CFR 862.3650 | |----------------------|-----------------------------------------------------------------------------------| | Common Name: | Oxycodone Enzyme Immunoassay | | Proprietary Name: | LZI Oxycodone III Enzyme Immunoassay | {4}------------------------------------------------ ### Legally Marketed Predicate Device(s) The LZI Oxycodone III Enzyme Immunoassay (EIA) is substantially equivalent to the LZI Oxycodone Assay (k120763) manufactured by Lin-Zhi International, Inc. (LZI). The LZI Oxycodone III 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 Oxycodone III Enzyme Immunoassay is a homogeneous enzyme immunoassay with ready-to-use liquid 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 oxycodone standard and referred to as oxycodone-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, oxycodone-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 oxycodone-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 Oxycodone III 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 Oxycodone III Enzyme Immunoassay is traceable to a commercially available oxycodone standard. The Ri solution contains mouse monoclonal anti-oxycodone 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 oxycodone in buffer with sodium azide (0.09 %) as a preservative. # Intended Use The LZI Oxycodone III Enzyme Immunoassay is intended for the qualitative and semiquantitative determination of oxycodone in human urine at the cutoff values of 100 ng/mL and 300 ng/mL when calibrated against oxycodone. 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 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 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. {5}------------------------------------------------ # Comparison to Predicate Device The LZI Oxycodone III Enzyme Immunoassay is substantially equivalent to the LZI Oxycodone Assay cleared by the FDA under the premarket notification k120763 for its stated intended use. | Device Characteristics | Subject Device | Predicate Device (k120763) | |---------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Intended Use | LZI Oxycodone III Enzyme Immunoassay The Lin-Zhi International, Inc. (LZI) Oxycodone III Enzyme Immunoassay is intended for the qualitative and semi- quantitative determination of oxycodone in human urine at a cutoff value of 100 ng/mL and 300 ng/mL when calibrated against oxycodone. 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 verification by a confirmatory method such as 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 confirmatory method (e.g., gas or liquid chromatography and mass spectrometry) must be used to obtain a confirmed analytical result (1, 2). Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. | LZI Oxycodone Enzyme Immunoassay The Lin-Zhi International, Inc. (LZI) Oxycodone Enzyme Immunoassay is intended for the qualitative and semi- quantitative determination of oxycodone in human urine, at cutoff values of 100 ng/mL and 300 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 analytical chemistry method must be used in order to obtain a confirmed analytical result. Gas or Liquid Chromatography/Mass Spectrometry (GC/MS or LC/MS) are the preferred confirmatory methods. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. | | Target Analyte | oxycodone | oxycodone | | Cutoff | 100 ng/ and 300 ng/mL | 100 and 300 ng/mL | | Matrix | Urine | Urine | | Calibrator Levels | 100 ng/mL Cutoff: 5 Levels 0, 50, 100, 150, and 300 ng/mL 300 ng/mL Cutoff: 5 Levels 0, 150, 300, 500, and 800 ng/mL | 0, 50, 100, 300, 500, and 800 ng/mL | The following table compares LZI Oxycodone III Enzyme Immunoassay with the predicate device. {6}------------------------------------------------ | Device Characteristics | Subject Device | Predicate Device (k120763) | |---------------------------|-------------------------------------------------|-------------------------------------------------| | Control Levels | LZI Oxycodone III Enzyme Immunoassay | LZI Oxycodone Enzyme Immunoassay | | | 100 ng/mL Cutoff: 2 Levels 75 and 125 ng/mL | 100 ng/mL Cutoff: 2 Levels 75 and 125 ng/mL | | | 300 ng/mL Cutoff: 2 Levels 225 and 375 ng/mL | 300 ng/mL Cutoff: 2 Levels 225 and 375 ng/mL | | | Storage | 2-8 °C until expiration date | # Performance Characteristics Summary: All validation studies below were conducted on the Beckman Coulter® AU480 Analyzer ### Precision: 100 ng/mL Cutoff The assay was tested in qualitative and semi-quantitative mode by spiking oxycodone into pooled negative urine at concentrations ±25 %, ±50 %, ±75 %, and ±100 % of the cutoff concentration. Results shown below were obtained by testing all samples in replicate of two runs a day for 22 days on one Beckman Coulter AU480 automatic clinical analyzer for a total of 88 runs. | 100 ng/mL Cutoff Result: | | Within Run (N=22) | | Total Precision (N=88) | | |--------------------------|-------------|-------------------|----------------|------------------------|-----------------| | Oxycodone | % of Cutoff | Number of | Immunoassay | Number of | Immunoassay | | Concentration | | Determinations | Result | Determinations | Result | | 0 ng/mL | 0 % | 22 | 22 Negative | 88 | 88 Negative | | 25 ng/mL | 25 % | 22 | 22 Negative | 88 | 88 Negative | | 50 ng/mL | 50 % | 22 | 22 Negative | 88 | 88 Negative | | 75 ng/mL | 75 % | 22 | 22 Negative | 88 | 88 Negative | | 100 ng/mL | 100 % | 22 | 5 Neg / 17 Pos | 88 | 26 Neg / 62 Pos | | 125 ng/mL | 125 % | 22 | 22 Positive | 88 | 88 Positive | | 150 ng/mL | 150 % | 22 | 22 Positive | 88 | 88 Positive | | 175 ng/mL | 175 % | 22 | 22 Positive | 88 | 88 Positive | | 200 ng/mL | 200 % | 22 | 22 Positive | 88 | 88 Positive | #### Semi-Quantitative Positive/Negative Results: #### Qualitative Positive/Negative Results: | 100 ng/mL Cutoff Result: | | Within Run (N=22) | | Total Precision (N=88) | | |--------------------------|-------------|-------------------|----------------|------------------------|-----------------| | Oxycodone | % of Cutoff | Number of | Immunoassay | Number of | Immunoassay | | Concentration | | Determinations | Result | Determinations | Result | | 0 ng/mL | 0 % | 22 | 22 Negative | 88 | 88 Negative | | 25 ng/mL | 25 % | 22 | 22 Negative | 88 | 88 Negative | | 50 ng/mL | 50 % | 22 | 22 Negative | 88 | 88 Negative | | 75 ng/mL | 75 % | 22 | 22 Negative | 88 | 88 Negative | | 100 ng/mL | 100 % | 22 | 9 Neg / 13 Pos | 88 | 33 Neg / 55 Pos | | 125 ng/mL | 125 % | 22 | 22 Positive | 88 | 88 Positive | | 150 ng/mL | 150 % | 22 | 22 Positive | 88 | 88 Positive | | 175 ng/mL | 175 % | 22 | 22 Positive | 88 | 88 Positive | | 200 ng/mL | 200 % | 22 | 22 Positive | 88 | 88 Positive | {7}------------------------------------------------ ## Analytical Recovery: 100 ng/mL Cutoff To demonstrate recovery of the entire assay range, a drug free-urine pool spiked with oxycodone at 300 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. Results are summarized below: | Target Concentration (ng/mL) | Determined Concentration Range (ng/mL) | Determined Concentration Average (ng/mL) | Average % Recovery | |------------------------------------|----------------------------------------------|------------------------------------------------|-----------------------| | 300 | 299.7 - 304.7 | 302.1 | 100.7 % | | 270 | 277.5 - 287.4 | 282.7 | 104.7 % | | 240 | 253.3 - 264.3 | 260.4 | 108.5 % | | 210 | 219.8 - 241.2 | 231.0 | 110.0 % | | 180 | 193.3 - 201.8 | 197.0 | 109.5 % | | 150 | 149.1 - 158.7 | 153.6 | 102.4 % | | 120 | 118.6 - 124.2 | 121.4 | 101.2 % | | 90 | 86.3 - 90.7 | 88.8 | 98.6 % | | 60 | 54.5 - 59.4 | 56.9 | 94.8 % | | 30 | 25.7 - 29.5 | 27.1 | 90.3 % | | 0 | -0.7 - 1.7 | 0.7 | N/A | ## Method Comparison - Clinical Samples: 100 ng/mL Cutoff A total of eighty-two (82) unaltered clinical samples were tested with the LZI Oxycodone III Enzyme Immunoassay on the Beckman Coulter AU480 automated clinical analyzer. All samples were tested in singlet. #### Semi-Quantitative Results: | Oxycodone Results 100 ng/mL Cutoff | Negative | < 50 % of the cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50 % below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50 % above the cutoff concentration) | High Positive (Greater than 50 % above the cutoff concentration) | % Agreement | |---------------------------------------------|----------|------------------------------------------------------------------|--------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------|----------------| | Positive | 0 | 0 | 2* | 12 | 25 | 90.2 % | | Negative | 20 | 9 | 10 | 4** | 0 | 95.1 % | Discordant samples determined when comparing LC/MS oxycodone and oxymorphone results with EIA results on the Beckman Coulter AU480 automated clinical analyser are shown in the table below. | Sample # | LC/MS Oxycodone (ng/mL) | LC/MS Oxymorphone (ng/mL) | Total Oxycodone + Oxymorphone LC/MS (ng/mL) | |-------------|-------------------------------|---------------------------------|---------------------------------------------------------| | 37* | 35.5 | 54.4 | 89.9 | | 38* | 46.4 | 44.6 | 91.0 | | 42** | 0.0 | 102.7 | 102.7 | | 43** | 2.2 | 104.8 | 107.0 | {8}------------------------------------------------ | Sample # | LC/MS Oxycodone (ng/mL) | LC/MS Oxymorphone (ng/mL) | Total Oxycodone + Oxymorphone LC/MS (ng/mL) | |-------------|-------------------------------|---------------------------------|---------------------------------------------------------| | 50** | 12.6 | 118.5 | 131.1 | | 52** | 36.9 | 101.8 | 138.7 | * Discordant between 50 % below cutoff and cutoff concentration (50 - 99.9 ng/mL) ** Discordant between cutoff and 50 % above cutoff concentration (100 - 149.9 ng/mL) #### Qualitative Accuracy Study: | Oxycodone Results 100 ng/mL Cutoff | Negative | < 50 % of the cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50 % below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50 % above the cutoff concentration) | High Positive (Greater than 50 % above the cutoff concentration) | % Agreement | |---------------------------------------------|----------|------------------------------------------------------------------|--------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------|----------------| | Positive | 0 | 0 | 2* | 12 | 25 | 90.2 % | | Negative | 20 | 9 | 10 | 4** | 0 | 95.1 % | Discordant samples determined when comparing LC/MS oxycodone and oxymorphone results with EIA results on the Beckman Coulter AU480 automated clinical analyser are shown in the table below. | Sample # | LC/MS Oxycodone (ng/mL) | LC/MS Oxymorphone (ng/mL) | Total Oxycodone + Oxymorphone LC/MS (ng/mL) | |-------------|-------------------------------|---------------------------------|---------------------------------------------------------| | 37* | 35.5 | 54.4 | 89.9 | | 38* | 46.4 | 44.6 | 91.0 | | 42* | 0.0 | 102.7 | 102.7 | | 43* | 2.2 | 104.8 | 107.0 | | 50* | 12.6 | 118.5 | 131.1 | | 52* | 36.9 | 101.8 | 138.7 | * Discordant between 50% below cutoff and cutoff concentration (50 - 99.9 ng/mL) ** Discordant between cutoff and 50% above cutoff concentration (100 - 149.9 ng/mL) ### Cross-reactivity: 100 ng/mL Cutoff 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 duplicates. The table below lists the concentration of each test compound that gave a response approximately equivalent to that of the cutoff calibrator (as positive) or the maximal concentration of the compound tested that gave a response below the response of the cutoff calibrator (as negative). Compounds tested at high concentration (100,000 ng/mL) with results below the cutoff value were listed as Not Detected (ND). Compounds tested below the high concentration (100,000 ng/mL) that gave a result below the cutoff value were given a "< %" value. {9}------------------------------------------------ #### Oxycodone and Major Metabolites: | Compound | Test Concentration (ng/mL) | % Cross- reactivity | |----------------|----------------------------------|------------------------| | Oxycodone | 100 | 100.00 % | | Oxymorphone | 100 | 100.00 % | | Noroxycodone | 25,000 | 0.40 % | | Noroxymorphone | 60,000 | 0.17 % | #### Structurally Related Compounds: | Compound | Test Concentration (ng/mL) | % Cross- reactivity | |----------------------------------|----------------------------------|------------------------| | 6-Acetylmorphine | 100,000 | ND | | Buprenorphine | 100,000 | ND | | Codeine | 100,000 | ND | | Codeine-6ß-D-Glucuronide | 100,000 | ND | | Dextromethorphan | 100,000 | ND | | Dihydrocodeine | 100,000 | ND | | Hydrocodone | 25,000 | 0.40 % | | Hydromorphone | 25,000 | 0.40 % | | Levorphanol | 100,000 | ND | | Morphine | 100,000 | ND | | Morphine-3ß-D-Glucuronide | 100,000 | ND | | Morphine-6ß-D-Glucuronide | 100,000 | ND | | Naloxone | 100,000 | ND | | Naloxone-3B-D-Glucuronide | 100,000 | ND | | Norbuprenorphine | 100,000 | ND | | Norcodeine | 100,000 | ND | | Norhydrocodone | 100,000 | ND | | Oxymorphone-3B-D- Glucuronide | 230 | 43.48 % | Structurally unrelated compounds were additionally spiked into pooled negative human urine to desired concentrations (as described below). These solutions were then split into three portions; one without oxycodone, and the remaining two that were further spiked with oxycodone standards to a final oxycodone concentration of 75 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). Samples were then evaluated against the cutoff calibrator in qualitative mode or the assay's calibration curve in semi-quantitative 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). Compounds tested below the high concentration (100,000 ng/mL) that gave a result below the cutoff value were given a "< %" value. {10}------------------------------------------------ | Compound | Test Concentration (ng/mL) | -25 % Oxycodone Cutoff (75 ng/mL) Result | +25 % Oxycodone Cutoff (125 ng/mL) Result | |--------------------------------------------------------|----------------------------------|------------------------------------------------|-------------------------------------------------| | Acetaminophen | 100,000 | Neg | Pos | | Acetylsalicylic Acid | 100,000 | Neg | Pos | | Amitriptyline | 100,000 | Neg | Pos | | Amlodipine Besylate | 100,000 | Neg | Pos | | Amoxicillin | 100,000 | Neg | Pos | | d-Amphetamine | 100,000 | Neg | Pos | | Atorvastatin | 20,000 | Neg | Pos | | Benzoylecgonine | 100,000 | Neg | Pos | | Bupropion | 100,000 | Neg | Pos | | Caffeine | 100,000 | Neg | Pos | | Carbamazepine | 100,000 | Neg | Pos | | Cetirizine | 100,000 | Neg | Pos | | Chlorpheniramine | 100,000 | Neg | Pos | | Chlorpromazine | 100,000 | Neg | Pos | | Clomipramine | 100,000 | Neg | Pos | | Desipramine | 100,000 | Neg | Pos | | Diphenhydramine | 100,000 | Neg | Pos | | Duloxetine | 100,000 | Neg | Pos | | Fentanyl | 100,000 | Neg | Pos | | Fluoxetine | 100,000 | Neg | Pos | | Fluphenazine | 100,000 | Neg | Pos | | Gabapentin | 100,000 | Neg | Pos | | Ibuprofen | 100,000 | Neg | Pos | | Imipramine | 100,000 | Neg | Pos | | Lisinopril | 100,000 | Neg | Pos | | Losartan | 10,000 | Neg | Pos | | Loratadine | 100,000 | Neg | Pos | | MDA (3,4- methylenedioxyamphetamine) | 100,000 | Neg | Pos | | MDEA | 100,000 | Neg | Pos | | MDMA (3,4- methylenedioxymethamphetamine) | 100,000 | Neg | Pos | | Meperidine | 100,000 | Neg | Pos | | Metformin | 100,000 | Neg | Pos | | Metoprolol | 100,000 | Neg | Pos | | Methadone | 100,000 | Neg | Pos | | d-Methamphetamine | 100,000 | Neg | Pos | | Nicotine | 100,000 | Neg | Pos | | Nortriptyline | 100,000 | Neg | Pos | | Omeprazole | 100,000 | Neg | Pos | | Oxazepam | 100,000 | Neg | Pos | | Phenobarbital | 100,000 | Neg | Pos | | (1S,2S)-(+)Pseudoephedrine | 100,000 | Neg | Pos | | Compound | Test Concentration (ng/mL) | -25 % Oxycodone Cutoff (75 ng/mL) Result | +25 % Oxycodone Cutoff (125 ng/mL) Result | | Ranitidine | 100,000 | Neg | Pos | | Salbutamol (Albuterol) | 100,000 | Neg | Pos | | Sertraline | 100,000 | Neg | Pos | | THC-COOH (11-Nor-Delta-9-THC-9- carboxylic acid) | 1000 | Neg | Pos | | l -Thyroxine | 10,000 | Neg | Pos | | Tramadol | 100,000 | Neg | Pos | | Zolpidem | 10,000 | Neg | Pos | ## Structurally Unrelated Pharmacological Compounds: 100 ng/mL Cutoff {11}------------------------------------------------ ## Endogenous and Preservative Compound Interference: 100 ng/mL Cutoff Endogenous and Preservative compounds were spiked into pooled negative human urine to desired concentrations. These solutions were then split into three portions; one without oxycodone, and the remaining two that were further spiked with oxycodone standards to a final oxycodone concentration of 75 ng/mL or 125 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). Samples were then evaluated against the cutoff calibrator in qualitative mode and the assay's calibration curve in semi-quantitative mode. All samples were tested in duplicates. | Interfering Substance | Concentration of Compound (mg/dL) | -25 % Oxycodone Cutoff (75 ng/mL) | +25 % Oxycodone Cutoff (125 ng/mL) | |--------------------------|-----------------------------------------|--------------------------------------|---------------------------------------| | Acetone | 1000 | Neg | Pos | | Ascorbic Acid | 1500 | Neg | Pos | | Bilirubin | 2 | Neg | Pos | | Boric Acid | 1000 | Neg | Neg | | Calcium Chloride (CaCl2) | 300 | Neg | Pos | | Citric Acid (pH 3) | 800 | Neg | Pos | | Creatinine | 500 | Neg | Pos | | Ethanol | 1000 | Neg | Pos | | Galactose | 10 | Neg | Pos | | y-Globulin | 500 | Neg | Pos | | Glucose | 3000 | Neg | Pos | | Hemoglobin | 300 | Neg | Pos | | β-hydroxybutyric Acid | 100 | Neg | Pos | | Human Serum Albumin | 500 | Neg | Pos | | Oxalic Acid | 100 | Neg | Pos | | Potassium Chloride | 6000 | Neg | Pos | | Riboflavin | 7.5 | Neg | Pos | | Urea | 6000 | Neg | Pos | | Uric Acid | 10 | Neg | Pos | | Sodium Azide | 1000 | Neg | Pos | | Sodium Chloride | 6000 | Neg | Pos | | Sodium Fluoride | 1000 | Neg | Pos | | Sodium Phosphate | 300 | Neg | Pos | {12}------------------------------------------------ The following endogenous and preservative compounds which showed interference at ±25 % of the cutoff concentration were then spiked into negative urine and at ±50 % of the cutoff concentration (50 ng/mL and 150 ng/mL) for the assay. Interference was observed with Boric Acid at 1 % w/v. No other significant undesired crossreactants or endogenous/preservative substance interference were observed. | Interfering Substance | Concentration of Compound (mg/dL) | -50 % Oxycodone Cutoff (50 ng/mL) | +50 % Oxycodone Cutoff (150 ng/mL) | |-----------------------|-----------------------------------------|--------------------------------------|---------------------------------------| | Boric Acid | 1000 | Neg | Neg | ## Specific Gravity Interference: 100 ng/mL Cutoff Samples ranging in specific gravity from 1.000 to 1.030 were spiked to a final oxycodone concentration of either 75 ng/mL or 125 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration. respectively). These samples were then evaluated in both semi-quantitative and qualitative modes. There were no deviations from the expected positive or negative results. # pH Interference: 100 ng/mL Cutoff Negative urine and urine spiked with oxycodone to a final oxycodone concentration of either 75 ng/mL or 125 ng/mL (as negative or positive controls. ±25 % of the cutoff concentration. respectively) were adjusted to pH levels from 3 to 11 and tested by the assay. The pH adjusted solutions were evaluated in both qualitative and semi-quantitative modes and there were no deviations from the expected results. # Precision: 300 ng/mL Cutoff The assay was tested in qualitative and semi-quantitative mode by spiking oxycodone into pooled negative urine at concentrations ±25 %, ±50 %, ±75 %, and ±100 % of the cutoff concentration. Results shown below were obtained by testing all samples in replicate of two runs a day for 22 days on one Beckman Coulter AU480 automatic clinical analyzer for a total of 88 runs. | 300 ng/mL Cutoff Result: | | Within Run (N=22) | | Total Precision (N=88) | | |----------------------------|-------------|----------------------------|-----------------------|----------------------------|-----------------------| | Oxycodone Concentration | % of Cutoff | Number of Determination | Immunoassay Result | Number of Determination | Immunoassay Result | | 0 ng/mL | 0 % | 22 | 22 Negative | 88 | 88 Negative | | 75 ng/mL | 25 % | 22 | 22 Negative | 88 | 88 Negative | | 150 ng/mL | 50 % | 22 | 22 Negative | 88 | 88 Negative | | 225 ng/mL | 75 % | 22 | 22 Negative | 88 | 88 Negative | | 300 ng/mL | 100 % | 22 | 6 Neg/16 Pos | 88 | 27 Neg/61 Pos | | 375 ng/mL | 125 % | 22 | 22 Positive | 88 | 88 Positive | | 450 ng/mL | 150 % | 22 | 22 Positive | 88 | 88 Positive | | 525 ng/mL | 175 % | 22 | 22 Positive | 88 | 88 Positive | | 600 ng/mL | 200 % | 22 | 22 Positive | 88 | 88 Positive | Semi-Ouantitative Positive/Negative Results: Qualitative Positive/Negative Results: {13}------------------------------------------------ | 300 ng/mL Cutoff Result: | | Within Run (N=22) | | Total Precision (N=88) | | |----------------------------|-------------|-----------------------------|-----------------------|-----------------------------|-----------------------| | Oxycodone Concentration | % of Cutoff | Number of Determinations | Immunoassay Result | Number of Determinations | Immunoassay Result | | 0 ng/mL | 0 % | 22 | 22 Negative | 88 | 88 Negative | | 75 ng/mL | 25 % | 22 | 22 Negative | 88 | 88 Negative | | 150 ng/mL | 50 % | 22 | 22 Negative | 88 | 88 Negative | | 225 ng/mL | 75 % | 22 | 22 Negative | 88 | 88 Negative | | 300 ng/mL | 100 % | 22 | 10 Neg / 12 Pos | 88 | 40 Neg / 48 Pos | | 375 ng/mL | 125 % | 22 | 22 Positive | 88 | 88 Positive | | 450 ng/mL | 150 % | 22 | 22 Positive | 88 | 88 Positive | | 525 ng/mL | 175 % | 22 | 22 Positive | 88 | 88 Positive | | 600 ng/mL | 200 % | 22 | 22 Positive | 88 | 88 Positive | # Analytical Recovery: 300 ng/mL Cutoff To demonstrate recovery of the entire assay range, a drug free-urine pool spiked with oxycodone at 800 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. Results are summarized below. | Target Concentration (ng/mL) | Determined Concentration Range (ng/mL) | Determined Concentration Average (ng/mL) | Average % Recovery | |------------------------------------|----------------------------------------------|------------------------------------------------|-----------------------| | 800 | 829.6 - 859.3 | 843.5 | 105.4 % | | 720 | 769.7 - 794.1 | 784.2 | 108.9 % | | 640 | 689.6 - 731.2 | 712.8 | 111.4 % | | 560 | 603.8 - 640.0 | 624.3 | 111.5 % | | 480 | 497.8 - 525.2 | 514.2 | 107.1 % | | 400 | 427.2 - 451.5 | 436.8 | 109.2 % | | 320 | 327.6 - 359.1 | 345.0 | 107.8 % | | 240 | 242.9 - 260.8 | 250.8 | 104.5 % | | 160 | 168.3 - 183.4 | 173.9 | 108.7 % | | 80 | 82.9 - 95.0 | 89.0 | 111.2 % | | 0 | -5.2 - 5.9 | 0.3 | N/A | ### Method Comparison - Clinical Samples: 300 ng/mL Cutoff A total of ninety (90) unaltered clinical samples were tested with the LZI Oxycodone III Enzyme Immunoassay on the Beckman Coulter AU480 automated clinical analyzer. All samples were tested in singlet. All samples were confirmed with LC/MS for oxycodone and oxymorphone concentrations. {14}------------------------------------------------ #### Semi-Quantitative Results: | Oxycodone Results 300 ng/mL Cutoff | Negative | < 50 % of the cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50 % below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50 % above the cutoff concentration) | High Positive (Greater than 50 % above the cutoff concentration) | % Agreement | |---------------------------------------------|----------|------------------------------------------------------------------|--------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------|----------------| | Positive | 0 | 0 | 2* | 11 | 33 | 97.8 % | | Negative | 20 | 6 | 17 | 1** | 0 | 95.6 % | Discordant samples determined when comparing LC/MS oxycodone and oxymorphone results with EIA results on the Beckman Coulter AU480 automated clinical analyser are shown in the table below. | Sample # | LC/MS Oxycodone (ng/mL) | LC/MS Oxymorphone (ng/mL) | Total Oxycodone + Oxymorphone LC/MS (ng/mL) | |-------------|-------------------------------|---------------------------------|---------------------------------------------------------| | 42* | 200.2 | 49.5 | 249.7 | | 43* | 53.0 | 203.5 | 256.5 | | 55** | 46.7 | 367.3 | 414.0 | * Discordant between 50 % below cutoff and cutoff concentration (150 - 299.9 ng/mL) ** Discordant between cutoff and 50 % above cutoff concentration (300 - 449.9 ng/mL) #### Qualitative Accuracy Study: | Oxycodone Results 300 ng/mL Cutoff | Negative | < 50 % of the cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50 % below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50 % above the cutoff concentration) | High Positive (Greater than 50 % above the cutoff concentration) | % Agreement | |---------------------------------------------|----------|------------------------------------------------------------------|--------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------|----------------| | Positive | 0 | 0 | 2* | 10 | 33 | 95.6 % | | Negative | 20 | 6 | 17 | 2** | 0 | 95.6 % | Discordant samples determined when comparing LC/MS oxycodone and oxymorphone results with EIA results on the Beckman Coulter AU480 automated clinical analyser are shown in the table below. | Sample # | LC/MS Oxycodone (ng/mL) | LC/MS Oxymorphone (ng/mL) | Total Oxycodone + Oxymorphone LC/MS (ng/mL) | |-------------|-------------------------------|---------------------------------|---------------------------------------------------------| | 42* | 200.2 | 49.5 | 249.7 | | 43* | 53.0 | 203.5 | 256.5 | | 50** | 177.2 | 212.1 | 389.3 | | 55** | 46.7 | 367.3 | 414.0 | * Discordant between 50% below cutoff and cutoff concentration (150 - 299.9 ng/mL) ** Discordant between cutoff and 50% above cutoff concentration (300 - 449.9 ng/mL) {15}------------------------------------------------ ### Cross-reactivity: 300 ng/mL Cutoff 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 duplicates. The table below lists the concentration of each test compound that gave a response approximately equivalent to that of the cutoff calibrator (as positive) or the maximal concentration of the compound tested that gave a response below the response of the cutoff calibrator (as negative). Compounds tested at high concentration (100,000 ng/mL) with results below the cutoff value were listed as Not Detected (ND). Compounds tested below the high concentration (100,000 ng/mL) that gave a result below the cutoff value were given a "< %" value. #### Oxycodone and Major Metabolites: | Compound | Test Concentration (ng/mL) | % Cross- reactivity | |----------------|----------------------------------|------------------------| | Oxycodone | 300 | 100.00 % | | Oxymorphone | 300 | 100.00 % | | Noroxycodone | 75,000 | 0.40 % | | Noroxymorphone | 100,000 | ND | #### Structurally Related Compounds: | Compound | Test Concentration (ng/mL) | % Cross- reactivity | |----------------------------------|----------------------------------|------------------------| | 6-Acetylmorphine | 100,000 | ND | | Buprenorphine | 100,000 | ND | | Codeine | 100,000 | ND | | Codeine-6β-D-Glucuronide | 100,000 | ND | | Dextromethorphan | 100,000 | ND | | Dihydrocodeine | 100,000 | ND | | Hydrocodone | 75,000 | 0.40 % | | Hydromorphone | 75,000 | 0.40 % | | Levorphanol | 100,000 | ND | | Morphine | 100,000 | ND | | Morphine-3β-D-Glucuronide | 100,000 | ND | | Morphine-6β-D-Glucuronide | 100,000 | ND | | Naloxone | 100,000 | ND | | Naloxone-3β-D-Glucuronide | 100,000 | ND | | Norbuprenorphine | 100,000 | ND | | Norcodeine | 100,000 | ND | | Norhydrocodone | 100,000 | ND | | Oxymorphone-3β-D- Glucuronide | 700 | 42.86 % | {16}------------------------------------------------ Structurally unrelated compounds were additionally spiked into pooled negative human urine to desired concentrations (as described below). These solutions were then split into three portions; one without oxycodone, and the remaining two that were further spiked with oxycodone standards to a final oxycodone concentration of 225 ng/mL or 375 ng/mL (as negative or positive controls, ±25 % of the cutoff concentration, respectively). Samples were then evaluated against the cutoff calibrator in qualitative mode and against the assay's calibration curve in semiquantitative 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). Compounds tested below the high concentration (100,000 ng/mL) that gave a result below the cutoff value were given a "< %" value. | Compound | Test Concentration (ng/mL) | -25 % Oxycodone Cutoff (225 ng/mL) Result | +25 % Oxycodone Cutoff (375 ng/mL) Result | |------------------------------------------|----------------------------------|-------------------------------------------------|-------------------------------------------------| | Acetaminophen | 100,000 | Neg | Pos | | Acetylsalicylic Acid | 100,000 | Neg | Pos | | Amitriptyline | 100,000 | Neg | Pos | | Amlodipine Besylate | 100,000 | Neg | Pos | | Amoxicillin | 100,000 | Neg | Pos | | d-Amphetamine | 100,000 | Neg | Pos | | Atorvastatin | 20,000 | Neg | Pos | | Benzoylecgonine | 100,000 | Neg | Pos | | Bupropion | 100,000 | Neg | Pos | | Caffeine | 100,000 | Neg | Pos | | Carbamazepine | 100,000…