Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on a homogeneous enzyme immunoassay technique and standard analytical methods, with no mention of AI or ML algorithms for data analysis or interpretation.

Therapeutic

No.
Explanation: This device is an in vitro diagnostic (IVD) assay designed to detect buprenorphine in human urine. It is used for analytical purposes (qualitative and semiquantitative analysis) and provides a preliminary test result, which needs confirmation by other methods. It does not provide any treatment or therapeutic benefit to a patient.

Diagnostic

Yes.

The "Intended Use / Indications for Use" section states, "The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay... For in vitro diagnostic use." This explicitly labels the device as intended for diagnostic use.

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic assay that uses chemical reagents and is intended for use with chemistry analyzers. It is not a software-only medical device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Explicit Statement: The document explicitly states "For in vitro diagnostic use."
Intended Use: The intended use is for the "qualitative and/or semiquantitative analyses of buprenorphine in human urine," which is a biological specimen. This analysis is performed in vitro (outside of the living body).
Device Description: The device description details a laboratory assay using reagents to analyze a biological sample (urine).
Professional Use: It is intended for use in "laboratories" by "Professional Use," which aligns with the typical use setting for IVDs.
Regulatory Statement: The caution about Federal (USA) law restricting sale by or on the order of a licensed healthcare professional is common for IVDs.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay with a 5 ng/mL cutoff. The assay is intended for use in laboratories for the qualitative and/or semiquantitative analyses of buprenorphine in human urine. Emit® II Plus assays are designed for use with a number of chemistry analyzers.

The semiquantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Mass Spectrometry (LC/MS) or permitting laboratories to establish quality control procedures.

The Emit® II Plus Burrenorphine Assay provides only a preliminary analytical test result. A more specific alternative chemical method(s) must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/ MS) or LC/MS are the preferred confirmatory methods. Other chemical confirmation methods are available. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when preliminary positive results are used.

For Professional Use.

Caution: Federal (USA) law restricts this device to sale by or on the order of a licensed healthcare professional.

For in vitro diagnostic use.

Product codes (comma separated list FDA assigned to the subject device)

DJG

Device Description

The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay technique used for the analysis of specific compounds in human urine. The assay is based on competition between drug in the specimen and drug labeled with the recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, so the drug concentration in the specimen can be measured in terms of enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous serum G6PDH does not interfere because the coenzyme NAD functions only with the bacterial (Leuconostoc mesenteroides) enzyme employed in the assay.

The Emit® II Plus Buprenorphine Assay reagents are provided liquid, ready to use and may be used directly from the refrigerator. The product is sold in three (3) kit sizes: 28 mL, 115 mL, and 1000 mL. Reagents 1 and 2 are provided as a matched set. They should not be interchanged with components of kits with different lot numbers.

Antibody/Substrate Reagent 1: Mouse monoclonal antibodies to buprenorphine (0.53 µg/mL). NAD (6.9 mM), G6P (10.9 mM), bovine serum albumin, preservatives, and stabilizers.
The antibody titer and enzyme conjugate activity may vary from lot to lot.

Enzyme Reagent 2: Norbuprenorphine labeled with bacterial rG6PDH (0.50 µg/mL), HEPES buffer, bovine serum albumin, preservatives, and stabilizers, where the antibody titer and enzyme conjugate activity may vary from lot to lot.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Human urine

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Laboratories; Professional Use.

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

Method Comparison:
Sample Size: One hundred twenty (120) native urine individual patient samples.
Data Source: Collected from external suppliers, received frozen and thawed before testing. Samples were distributed across the assay range, with 21% within -50% of the cutoff and 19% within +50% of the cutoff.
Annotation Protocol: Each specimen was tested on the DxC 500 AU and on LC/MS/MS. Results were analyzed using a 2x2 box plot to assess qualitative agreement relative to the 5 ng/mL cutoff value.

Recovery:
Sample Size: Nine (9) urine samples.
Data Source: Prepared by spiking buprenorphine into negative urine pools at levels of 2.0, 3.0, 4.0, 5.0, 8.0, 12.0, 18.0, 22.0, and 25.0 ng/mL.
Annotation Protocol: Tested in five (5) replicates using one reagent lot, one calibrator lot, one instrument, and one calibration. Samples were additionally analyzed by LC/MS/MS to determine the reference value.

Precision (Repeatability and Within-Lab Precision):
Sample Size: Eleven (11) urine samples.
Data Source: Prepared by spiking buprenorphine into negative urine pools, aliquoted and frozen, at levels of 0, 1.25, 2.5, 3.0, 3.75, 5.0, 6.25, 7.0, 7.5, 8.75, and 10.0 ng/mL.
Annotation Protocol: 20x2x2 design using one reagent lot, one calibrator lot, one instrument, one calibration, twenty test days, and two runs per test day with two replicate measurements each. Two runs were performed each day with a minimum of two hours in between.

Precision (Reproducibility Precision):
Sample Size: Three (3) urine samples (Negative Control (3 ng/mL), Cutoff Calibrator, Positive Control (7 ng/mL)).
Data Source: Emit II Plus Specialty Drug Control Negative, Emit II Plus Specialty Drug Cal/Ctrl, and Emit II Plus Specialty Drug Control Positive.
Annotation Protocol: 3x5x5 design; 3 (Sites/Instruments) x 5 (replicates per day) for a total of seventy-five (75) replicates per reagent lot. Three (3) Emit II Plus Buprenorphine reagent lots were processed in parallel. Testing completed by three (3) operators. Each testing day, one (1) run was performed on three (3) instruments.

Limit of Detection:
Sample Size: Four (4) blank samples and four (4) low-level samples (LoD).
Data Source: Blank samples were drug-free negative urine. Low samples were drug-free urine spiked with buprenorphine.
Annotation Protocol: Tested using two reagent lots, one calibrator lot, one instrument, one calibration, three testing days. Minimum of sixty (60) replicate measurements across all blank samples, days, and instrument. Minimum of sixty (60) replicate measurements across all low-level samples, days, and instrument.

Specificity and Cross-reactivity:
Sample Size: One (1) sample pool for each compound tested.
Data Source: Not explicitly stated, but implies spiked samples for metabolites, structurally related, and structurally unrelated compounds.
Annotation Protocol: Tested using one reagent lot, one calibrator lot, one instrument. Samples were assayed in five (5) replicates.

Interference:
Sample Size: Not explicitly stated, but implies multiple test samples for each interferent.
Data Source: Urine testing samples prepared by spiking exogenous materials (metabolites and structurally related compounds) into urine samples at ± 40% positive and negative control levels (3 ng/mL and 7 ng/mL respectively).
Annotation Protocol: One reagent lot, one calibrator lot, one instrument, one sample pool for each compound tested. Samples assayed in five (5) replicates.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Method Comparison

Study Type: Method comparison, qualitative analysis
Sample Size: 120 native urine individual patient samples
Standalone Performance: Qualitative analysis of Emit II Plus Buprenorphine on DxC 500 AU compared to LC/MS/MS
Key results: 92.5% Agreement.
- 48 samples were negative by both Emit II Plus Buprenorphine Assay and LC/MS/MS.
- 63 samples were positive by both Emit II Plus Buprenorphine Assay and LC/MS/MS.
- 9 discordant samples were observed as positive by Emit II Plus Buprenorphine and negative by LC/MS/MS.

Recovery

Study Type: Recovery, semi-quantitative analysis
Sample Size: 9 urine samples spiked with buprenorphine (2.0-25.0 ng/mL)
Standalone Performance: % Recovery vs Nominal and % Recovery vs LC/MS/MS
Key results: All samples showed recovery percentages ranging from 86% to 103% against nominal values and 86% to 100% against LC/MS/MS values.

Repeatability and Within-Lab Precision

Study Type: Precision study (qualitative and semi-quantitative analysis)
Sample Size: 11 urine samples spiked with buprenorphine (0-10.0 ng/mL)
Standalone Performance: Mean (mA/min) and Pos/Neg results for qualitative analysis, and Mean (ng/mL), SD, %CV for semi-quantitative analysis.
Key results:
- Qualitative Analysis: For concentrations below cutoff (0.0 to 3.75 ng/mL), all 80 results were Negative. For concentrations above cutoff (6.25 to 10.0 ng/mL), all 80 results were Positive. At cutoff (5.0 ng/mL), 21 Negative and 59 Positive results were observed.
- Semi-Quantitative Analysis: %CV for repeatability ranged from 1.1% to 6.6%. %CV for within-lab precision ranged from 2.8% to 6.6%.

Reproducibility Precision

Study Type: Reproducibility precision
Sample Size: 3 control samples (Negative Control, Cutoff Calibrator, Positive Control) with 225 replicates each.
Standalone Performance: Total Reproducibility Precision (Mean and %CV) and breakdown by Repeatability, Between Day, Between Lot, Between Instrument/Site.
Key results:
- Total Reproducibility Precision: Negative Control (Mean 3.1 ng/mL, %CV 5.3), Cutoff Calibrator (Mean 5.0 ng/mL, %CV 3.7), Positive Control (Mean 7.1 ng/mL, %CV 3.0).
- Component %CVs were relatively low, indicating good reproducibility across different factors.

Limit of Detection

Study Type: Limit of Detection
Sample Size: Blank and low-level spiked samples.
Key results: Information about the actual LoD values for Lot 1, Lot 2, Final, and Claimed Limit of Detection were present in the table but the values were not filled in the provided text.

Specificity and Cross-reactivity

Study Type: Specificity and Cross-reactivity
Standalone Performance: % Cross-reactivity for buprenorphine metabolites and structurally related/unrelated compounds.
Key results:
- Buprenorphine: 98% cross-reactivity.
- Norbuprenorphine: 106% cross-reactivity.
- Buprenorphine Glucuronide and Norbuprenorphine Glucuronide: 0.10% cross-reactivity at 1,000 ng/mL.
- Structurally related compounds (e.g., 6-Acetylcodeine, Morphine, Codeine): All showed

Regulation Number and Section

§ 862.3650 Opiate test system.

(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (e.g., programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).

Summary

0

Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.

Siemens Healthcare Diagnostics Inc. Julie Warren Regulatory Affairs Professional 500 GBC Dr. P.O. Box 6101, Mailstop 514 Newark, DE 19714

Re: K221605

Trade/Device Name: Emit® II Plus Buprenorphine Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate Test System Regulatory Class: Class II Product Code: DJG Dated: March 16, 2023 Received: March 17, 2023

Dear Julie Warren:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR

1

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 medical devices and radiation-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.

Joseph A. Digitally signed by Kotarek -S Date: 2023.07.25

Joseph Kotarek, Ph.D. Branch Chief Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K221605

Device Name Emit® II Plus Buprenorphine Assay

Indications for Use (Describe)

The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay with a 5 ng/mL cutoff. The assay is intended for use in laboratories for the qualitative and/or semiquantitative analyses of buprenorphine in human urine. Emit® II Plus assays are designed for use with a number of chemistry analyzers.

The semiquantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Mass Spectrometry (LC/MS) or permitting laboratories to establish quality control procedures.

The Emit® II Plus Burrenorphine Assay provides only a preliminary analytical test result. A more specific alternative chemical method(s) must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/ MS) or LC/MS are the preferred confirmatory methods. Other chemical confirmation methods are available. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when preliminary positive results are used.

For Professional Use.

Caution: Federal (USA) law restricts this device to sale by or on the order of a licensed healthcare professional.

For in vitro diagnostic use.

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

Image /page/3/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the words is a graphic of orange dots arranged in a circular pattern.

This 510(k) Summary of Safety and Effectiveness is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K221605

1. Submitter

Siemens Healthcare Diagnostics Inc. 500 GBC Dr. P.O. Box 6101 Newark, DE 19714

Primary Contact Person: Julie Warren Date of Preparation: July 7, 2023

2. Device Information

Trade Name	Emit® II Plus Buprenorphine Assay
Common Name	Buprenorphine Assay
Device	Enzyme Immunoassay, Opiates
Review Panel	Toxicology
Product Code	DJG
Submission Type	510(k)
Regulation Number	21 CFR § 862.3650
Device Class	2

3. Purpose of Submission

The purpose of this premarket notification is for the Emit® II Plus Buprenorphine Assay on the DxC 500 AU Clinical Chemistry Analyzer.

4. Predicate Device

Predicate Device Name: Emit® II Plus Buprenorphine Assay 510(k) Number: K150606

4

Image /page/4/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange. To the right of the word "Healthineers" is a graphic of orange dots.

5. Device Description

Emit® II Plus Buprenorphine Assay

The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay technique used for the analysis of specific compounds in human urine. The assay is based on competition between drug in the specimen and drug labeled with the recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, so the drug concentration in the specimen can be measured in terms of enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous serum G6PDH does not interfere because the coenzyme NAD functions only with the bacterial (Leuconostoc mesenteroides) enzyme employed in the assay.

The Emit® II Plus Buprenorphine Assay reagents are provided liquid, ready to use and may be used directly from the refrigerator. The product is sold in three (3) kit sizes: 28 mL, 115 mL, and 1000 mL. Reagents 1 and 2 are provided as a matched set. They should not be interchanged with components of kits with different lot numbers.

Antibody/Substrate Reagent 1

Mouse monoclonal antibodies to buprenorphine (0.53 µg/mL)*.NAD (6.9 mM), G6P (10.9 mM), bovine serum albumin, preservatives, and stabilizers.

*The antibody titer and enzyme conjugate activity may vary from lot to lot.

Enzyme Reagent 2

Norbuprenorphine labeled with bacterial rG6PDH (0.50 µg/mL), HEPES buffer, bovine serum albumin, preservatives, and stabilizers, where the antibody titer and enzyme conjugate activity may vary from lot to lot.

5

Image /page/5/Picture/0 description: The image shows the Siemens Healthineers logo. The word "SIEMENS" is in teal, and the word "Healthineers" is in orange. To the right of the words is a series of orange dots that are arranged in a pattern that resembles an arrow pointing to the right.

6. Intended Use/Indications for Use

Emit® II Plus Buprenorphine Assay

The Emit® II Plus Buprenorphine Assay is a homogeneous enzyme immunoassay with a 5 ng/mL cutoff. The assay is intended for use in laboratories for the qualitative and/or semiquantitative analyses of buprenorphine in human urine. Emit® II Plus assays are designed for use with a number of chemistry analyzers.

The semiquantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/Mass Spectrometry (LC/MS) or permitting laboratories to establish quality control procedures.

The Emit® II Plus Buprenorphine Assay provides only a preliminary analytical test result. A more specific alternative chemical method(s) must be used to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or LC/MS are the preferred confirmatory methods. Other chemical confirmation methods are available. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when preliminary positive results are used.

For Professional Use.

Caution: Federal (USA) law restricts this device to sale by or on the order of a licensed healthcare professional.

For in vitro diagnostic use.

7. Comparison of Technological Characteristics with the Predicate Device

The similarities and differences between the proposed device Emit II Plus Buprenorphine Assay on Beckman Coulter DxC AU 500 Clinical Chemistry Analyzer and the predicate device, Emit II Plus Buprenorphine Assay, were compared side by side.

| Feature | Predicate Device:
Emit II Plus Buprenorphine
Assay | Proposed Device:
Emit II Plus Buprenorphine
Assay |
|--------------|-------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------|
| Intended Use | A homogeneous enzyme
immunoassay with a 5
ng/mL cutoff. The assay is
intended for use in
laboratories for the
qualitative and/or | Same |

6

Image /page/6/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the words is a graphic of orange dots arranged in a pattern.

| Feature | Predicate Device:
Emit II Plus Buprenorphine
Assay | Proposed Device:
Emit II Plus Buprenorphine
Assay |
|--------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------|
| | semiquantitative analyses of
buprenorphine in human
urine. Preliminary analytical
test result. | |
| Analyte | Buprenorphine | Same |
| Cutoff Level | 5 ng/mL | Same |
| Measurement | Qualitative, semi-
quantitative | Same |
| Methodology | EMIT® technology:
Homogeneous enzyme
immunoassay | Same |
| Detection | Absorbance change
measured
Spectrophotometrically at
340nm | Same |
| Instrument | Analyzers must be capable
of maintaining a constant
reaction temperature,
pipette specimens/reagents,
mix thoroughly, measure
enzyme rates precisely and
time the reaction accurately. | Beckman Coulter DxC 500
AU Analyzer |
| Sample Type | Human Urine | Same |
| Reagent form and storage | Liquid - ready to use, on-
board storage | Same |
| Calibrator | 0, 2.5, 5, 15, and 25 ng/mL | Same |
| Control Levels | Positive: 7 ng/mL
Negative: 3 ng/mL | Same |

8. Performance Data

The following performance studies were conducted to demonstrate substantial equivalence of the Emit II Plus Buprenorphine Assay on the DXC 500 AU Clinical Chemistry Analyzer to the Emit II Plus Buprenorphine Assay previously cleared under premarket notification 510(k), K150606.

7

Image /page/7/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the words is a graphic of orange dots arranged in a circular pattern.

8.1 Method Comparison

The method comparison study was evaluated in accordance with CLSI EP09c 3rd Edition Measurement Procedure Comparison and Bias Estimation Using Patient Samples Evaluation of Precision Performance of Quantitative Measurement Methods and CLSI EP 12-A2 User Protocol for Evaluation of Qualitative Test Performance to determine accuracy and verification of the 5 ng/mL cutoff.

One hundred twenty (120) native urine individual patient samples were collected from external suppliers. The samples were received frozen, and thawed before testing. The samples were distributed across the assay range, 21% of which were within -50% of the cutoff and 19% of which were within +50% of the cutoff.

Each specimen was tested on the DxC 500 AU and on LC/MS/MS The results were analyzed using a 2x2 box plot to assess qualitative agreement relative to the 5 ng/mL cutoff value.

		LC/MS/MS
		Neg	Pos
Emit II Plus
Buprenorphine on DxC
500 AU	Neg	48	0
	Pos	9	63

Table 1. Method Comparison - Qualitative Analysis

Result: 92.5% Agreement

Table 2. Method Comparison - Discordant Results

| Sample
ID | DxC
500 AU
(ng/mL) | Buprenorphine
(ng/mL) | Norbuprenorphine
(ng/mL) | Buprenorphine+
Norbuprenorphine
(ng/mL) | DxC 500
AU
Pos/Neg | LC/MS/MS
Pos/Neg |
|--------------|--------------------------|--------------------------|-----------------------------|-----------------------------------------------|--------------------------|---------------------|
| BUP35 | 6.1 | 0.0 | 4.2 | 4.2 | Pos | Neg |
| BUP45 | 6.0 | 0.0 | 4.6 | 4.6 | Pos | Neg |
| BUP51 | 6.1 | 0.0 | 4.3 | 4.3 | Pos | Neg |
| BUP60 | 5.6 | 0.0 | 3.8 | 3.8 | Pos | Neg |
| BUP62 | 6.1 | 0.0 | 3.9 | 3.9 | Pos | Neg |
| BUP87 | 5.5 | 0.0 | 4.3 | 4.3 | Pos | Neg |
| BUP95 | 5.6 | 0.0 | 4.5 | 4.5 | Pos | Neg |
| BUP101 | 6.3 | 0.0 | 4.8 | 4.8 | Pos | Neg |
| BUP109 | 5.0 | 0.0 | 4.3 | 4.3 | Pos | Neg |

Emit® II Plus Buprenorphine Assay Traditional 510(k)

8

Image /page/8/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the word "Healthineers" is a cluster of orange dots.

One hundred twenty (120) samples were tested. Relative to the 5 ng/mL cutoff, fortyeight (48) samples were negative by Emit II Plus Buprenorphine Assay and LC/MS/MS, and sixty-three (63) were positive. Nine (9) discordant samples were observed as positive by Emit II Plus Buprenorphine and negative by LC/MS/MS.

8.2 Recovery

The recovery was evaluated in accordance with the following testing protocol. Nine (9) urine samples were prepared by spiking buprenorphine into negative urine pools. The following levels were tested: 2.0, 3.0, 4.0, 5.0, 8.0, 12.0, 18.0, 22.0, and 25.0 ng/mL.

Experimental design was (1) reagent lot, one (1) calibrator lot, one (1) instrument, one (1) calibration with two (2) replicate measurements each. Samples were assayed in five (5) replicates. Samples were additionally analyzed by LC/MS/MS to determine the reference value. The nominal value was determined gravimetrically based on the stock concentration.

| Sample
ID | Nominal
Value
(ng/mL) | DxC 500 AU
Value
(ng/mL) | %
Recovery
vs Nominal | LC/MS/MS
(ng/mL) | % Recovery
vs
LC/MS/MS |
|--------------|-----------------------------|--------------------------------|-----------------------------|---------------------|------------------------------|
| BUP1 | 2.0 | 1.9 | 95 | 2.2 | 86 |
| BUP2 | 3.0 | 2.9 | 97 | 3.2 | 91 |
| BUP3 | 4.0 | 4.1 | 103 | 4.4 | 93 |
| BUP4 | 5.0 | 4.9 | 98 | 5.1 | 96 |
| BUP5 | 8.0 | 7.6 | 95 | 8.8 | 86 |
| BUP6 | 12.0 | 11.6 | 97 | 13.1 | 89 |
| BUP7 | 18.0 | 18.1 | 101 | 18.1 | 100 |
| BUP8 | 22.0 | 22.9 | 104 | 22.9 | 100 |
| BUP9 | 25.0 | 24.7 | 99 | 25.6 | 96 |

Table 3. Recovery of Emit II Plus Buprenorphine - Semi-Quantitative Analysis

8.3 Precision

Repeatability and Within-Lab Precision

The precision was evaluated in accordance with CLSI EP05-A3 Evaluation of Precision Performance of Quantitative Measurement Methods.

Eleven (11) urine samples were prepared by spiking buprenorphine into negative urine pools, aliquoted and frozen prior to the start of testing. The following levels were tested: 0, 1.25, 2.5, 3.0, 3.75, 5.0, 6.25, 7.0, 7.5, 8.75, and 10.0 ng/mL.

Experimental design was a 20x2x2 design using (1) reagent lot, one (1) calibrator lot, one (1) instrument, one (1) calibration, twenty (20) test days and two (2) runs per test

9

Image /page/9/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the word "Healthineers" is a graphic of orange dots arranged in a circular pattern.

day with two (2) replicate measurements each. Each day, two runs were performed, with a minimum of two (2) hours in between.

| Sample
Concentration
(ng/mL) | % of the
Cutoff | # of
Results | Results
(Pos/Neg) | Mean
(mA/min) |
|------------------------------------|--------------------|-----------------|----------------------------|------------------|
| 0.0 | -100% | 80 | 80 Negative | 280 |
| 1.25 | -75% | 80 | 80 Negative | 297 |
| 2.5 | -50% | 80 | 80 Negative | 316 |
| 3.0 | -40% | 80 | 80 Negative | 323 |
| 3.75 | -25% | 80 | 80 Negative | 333 |
| 5.0 | 0% | 80 | 21 Negative
59 Positive | 363 |
| 6.25 | +25% | 80 | 80 Positive | 380 |
| 7.0 | +40% | 80 | 80 Positive | 392 |
| 7.5 | +50% | 80 | 80 Positive | 402 |
| 8.75 | +75% | 80 | 80 Positive | 427 |
| 10.0 | +100% | 80 | 80 Positive | 444 |

Table 4. Precision of Emit II Plus Buprenorphine Qualitative Analysis

Table 5. Precision of Emit II Plus Buprenorphine Semi-Quantitative Analysis

			Repeatability		Within-Lab Precision
Sample
Concentration
(ng/mL)	% of the
Cutoff	# of
Results	Results
(Pos/Neg)	Mean
(ng/mL)	SD	%CV	SD	%CV
0.0	-100%	80	80 Negative	0.1	0.06	-	0.11	-
1.25	-75%	80	80 Negative	1.2	0.08	6.6	0.08	6.6
2.5	-50%	80	80 Negative	2.3	0.09	3.7	0.11	4.6
3.0	-40%	80	80 Negative	2.7	0.07	2.8	0.15	5.7
3.75	-25%	80	80 Negative	3.3	0.08	2.5	0.15	4.4
5.0	0%	80	24 Negative
56 Positive	5.1	0.08	1.6	0.18	3.5
6.25	+25%	80	80 Positive	6.1	0.08	1.3	0.21	3.4
7.0	+40%	80	80 Positive	6.8	0.11	1.7	0.24	3.5
7.5	+50%	80	80 Positive	7.3	0.08	1.1	0.21	2.9
8.75	+75%	80	80 Positive	8.8	0.10	1.1	0.24	2.8
10.0	+100%	80	80 Positive	9.7	0.10	1.1	0.37	3.8

Reproducibility Precision

The reproducibility was evaluated in accordance with the following protocol.

Three (3) urine samples were tested: Emit II Plus Specialty Drug Control Negative (3 ng/mL) Emit II Plus Specialty Drug Cal/Ctrl (Cutoff Calibrator), and the Emit II Plus Specialty Drug Control Positive (7 ng/mL).

10

Image /page/10/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the word "Healthineers" is a cluster of orange dots arranged in a circular pattern.

Experimental design was a 3x5x5 design; 3 (Sites/Instruments) x 5 (replicates per day) for a total seventy-five (75) replicates per reagent lot. To capture lot-to-lot variability, three (3) Emit II Plus Buprenorphine reagent lots were processed in parallel in this study design. Testing was completed by three (3) operators. Each testing day, one (1) run was performed on three (3) instruments.

| Sample | N | Mean
(ng/mL) | SD | %CV |
|-------------------|-----|-----------------|------|-----|
| Negative Control | 225 | 3.1 | 0.17 | 5.3 |
| Cutoff Calibrator | 225 | 5.0 | 0.18 | 3.7 |
| Positive Control | 225 | 7.1 | 0.21 | 3.0 |

Table 6. Total Reproducibility Precision of Emit II Plus Buprenorphine Assay

	Table 7. Reproducibility Precision of Emit II Plus Buprenorphine Assay
--	------------------------------------------------------------------------	--

			Repeatability		Between Day		Between Lot		Between Instrument/Site
Sample	N	Mean (ng/mL)	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Negative Control	225	3.1	0.11	3.6	0.06	1.8	0.03	0.8	0.11	3.4
Cutoff Calibrator	225	5.0	0.11	2.2	0.07	1.4	0.00	0.0	0.13	2.6
Positive Control	225	7.1	0.12	1.7	0.08	1.2	0.05	0.7	0.15	2.1

8.4 Limit of Detection

The limits of detection, Limit of Blank (LoB) and Limit of Detection (LoD) were evaluated in accordance with CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures.

Blank samples were drug-free negative urine, and the low samples were drug-free urine spiked with buprenorphine.

Experimental design was (2) reagent lots, one (1) calibrator lot, one (1) instrument, one (1) calibration, three (3) testing days, four (4) blank samples with minimum of sixty (60) replicate measurements (across all blank samples, days, instrument) and four (4) samples, each containing a low level of measurand (LoD) with minimum of sixty (60) replicate measurements (across all low-level samples, days, instrument).

11

Image /page/11/Picture/0 description: The image shows the logo for Siemens Healthineers. The word "SIEMENS" is written in teal, and the word "Healthineers" is written in orange below it. To the right of the words is a graphic of orange dots arranged in a circular pattern.

Table 8. Limit of Detection Summary

| Sample | Limit of
Detection
Lot 1 (ng/mL) | Limit of
Detection
Lot 2
(ng/mL) | Limit of
Detection
Final (ng/mL) | Claimed
Limit of
Detection
(ng/mL) |
|--------|----------------------------------------|-------------------------------------------|----------------------------------------|---------------------------------------------|
| LoD | | | | |

8.5 Specificity and Cross-reactivity

The specificity was evaluated in accordance with CLSI EP07-ED3 Interference Testing Clinical Chemistry.

The experimental design was one (1) reagent lot, one (1) calibrator lot, one (1) instrument, one (1) sample pool for each compound tested. The samples were assayed in five (5) replicates.

Table 9. Specificity - Buprenorphine Metabolites

| Compound | Concentration
Tested (ng/mL) | Mean
(ng/mL) | Cutoff
(ng/mL) | % Cross-
reactivity |
|---------------------------------|---------------------------------|-----------------|-------------------|------------------------|
| Buprenorphine | 5 ng/mL | 4.9 | 5.0 | 98% |
| Norbuprenorphine | 5 ng/mL | 5.3 | 5.0 | 106% |
| Buprenorphine
Glucuronide | 1,000 ng/mL | 1.4 | 5.0 | 0.10% |
| Norbuprenorphine
Glucuronide | 1,000 ng/mL | 1.4 | 5.0 | 0.10% |

Table 10. Cross-reactivity - Structurally Related Compounds

| Description | Concentration
@ Testing | Mean
Rate
(mA/min) | Cutoff
Rate
(mA/min) | Response
vs. Cutoff
(Pos/Neg) | Mean
(ng/mL) | %Cross-
reactivity to 5
ng/mL Cutoff |
|------------------|----------------------------|--------------------------|----------------------------|-------------------------------------|-----------------|--------------------------------------------|
| 6-Acetylcodeine | 100,000 ng/mL | 131 | 167 | Neg | -0.6 |