Tina-quant Transferrin ver.2 (urine application) assay is an in vitro test for the quantitative determination of transferrin in human urine on Roche/Hitachi cobas c systems.

A transferrin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the transferrin (an iron-binding and transporting serum protein) in urine. Measurement of transferrin levels aids in the diagnosis of malnutrition, acute inflammation, infection, and red blood cell disorders, such as iron deficiency anemia.

Device Description

The Tina-quant Transferrin ver.2 (urine application) assay is a two reagent assay for the in vitro quantitative determination of transferrin in human urine on automated clinical chemistry analyzers. It is an immunoturbidimetric assay in which human transferrin forms a precipitate with a specific antiserum which is determined turbidimetrically.

Engineering drawings, schematics, and figures are not pertinent to describe the device, as the device is a reagent.

AI/ML Overview

The document provided is a 510(k) Premarket Notification for an in vitro diagnostic device, the "Tina-quant Transferrin ver.2 (urine application) assay." This type of submission focuses on demonstrating substantial equivalence to a legally marketed predicate device, rather than proving clinical effectiveness through extensive human studies often seen with novel medical devices. Therefore, the information regarding acceptance criteria and study design will be primarily focused on analytical performance validation rather than multi-reader multi-case clinical studies involving human interpretation of images, as this is a laboratory reagent.

Here's an analysis of the provided text in the context of your request:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria for this device are largely implicit in the fact that "All data passed the predetermined acceptance criteria" for each analytical study. The performance is reported as the results of these studies.

Performance Characteristic	Acceptance Criteria (Implicit: "All data passed the predetermined criteria")	Reported Device Performance
Precision	CV% and SD values within predefined limits.	Repeatability (within-run precision)
- PreciControl ClinChem Multi 1	Not explicitly stated, but results passed.	Mean: 1.98 mg/dL, SD: 0.0140 mg/dL, CV: 0.7%
- PreciControl ClinChem Multi 2	Not explicitly stated, but results passed.	Mean: 3.05 mg/dL, SD: 0.0242 mg/dL, CV: 0.8%
- Human Urine 1	Not explicitly stated, but results passed.	Mean: 0.435 mg/dL, SD: 0.00979 mg/dL, CV: 2.3%
- Human Urine 2	Not explicitly stated, but results passed.	Mean: 0.737 mg/dL, SD: 0.00920 mg/dL, CV: 1.2%
- Human Urine 3	Not explicitly stated, but results passed.	Mean: 1.27 mg/dL, SD: 0.0107 mg/dL, CV: 0.8%
- Human Urine 4	Not explicitly stated, but results passed.	Mean: 2.52 mg/dL, SD: 0.0184 mg/dL, CV: 0.7%
- Human Urine 5	Not explicitly stated, but results passed.	Mean: 3.30 mg/dL, SD: 0.0252 mg/dL, CV: 0.8%
Intermediate Precision (within-lab precision)	Not explicitly stated, but results passed.
- PreciControl ClinChem Multi 1	Not explicitly stated, but results passed.	Mean: 1.98 mg/dL, SD: 0.0158 mg/dL, CV: 0.8%
- PreciControl ClinChem Multi 2	Not explicitly stated, but results passed.	Mean: 3.05 mg/dL, SD: 0.0267 mg/dL, CV: 0.9%
- Human Urine 1	Not explicitly stated, but results passed.	Mean: 0.435 mg/dL, SD: 0.0111 mg/dL, CV: 2.5%
- Human Urine 2	Not explicitly stated, but results passed.	Mean: 0.737 mg/dL, SD: 0.0112 mg/dL, CV: 1.5%
- Human Urine 3	Not explicitly stated, but results passed.	Mean: 1.23 mg/dL, SD: 0.0130 mg/dL, CV: 1.1%
- Human Urine 4	Not explicitly stated, but results passed.	Mean: 2.52 mg/dL, SD: 0.0215 mg/dL, CV: 0.9%
- Human Urine 5	Not explicitly stated, but results passed.	Mean: 3.30 mg/dL, SD: 0.0289 mg/dL, CV: 0.9%
Analytical Sensitivity
- Limit of Blank (LoB)	LoB Claim: 0.10 mg/dL (highest measurement for blank sample with stated prob.)	Lot #1: 0.0150 mg/dL, Lot #2: 0.0130 mg/dL, Lot #3: 0.0150 mg/dL
- Limit of Detection (LoD)	LoD Claim: 0.15 mg/dL (lowest detectable analyte concentration with 95% prob.)	Lot #1: 0.0382 mg/dL, Lot #2: 0.0331 mg/dL, Lot #3: 0.0353 mg/dL
- Limit of Quantitation (LoQ)	LoQ Claim: 0.22 mg/dL with 20% CV (lowest quantifiable concentration)	Lot #1: 0.124 mg/dL, Lot #2: 0.137 mg/dL, Lot #3: 0.143 mg/dL
Linearity/Assay Reportable Range	Linear relationship across the measuring range.
- Measuring Range Claim	0.22 to 3.5 mg/dL.	Confirmed.
- Pearson Correlation Coefficient (r)	Close to 1.	Lot 1: 0.9999, Lot 2: 0.9999, Lot 3: 0.9997
Endogenous Interferences	No interference at specified concentrations.
- Albumin	No interference ≤ 5000 mg/L.	Passed (tested at 5 g/L).
- Calcium	No interference ≤ 8 mmol/L.	Passed (tested up to 9.92/9.80 mmol/L).
- Citrate	No interference ≤ 10 mmol/L.	Passed (tested up to 11 mmol/L).
- Creatinine	No interference ≤ 44 mmol/L.	Passed (tested up to 88 mmol/L).
- Glucose	No interference ≤ 111 mmol/L.	Passed (tested up to 388 mmol/L).
- Hemoglobin	No significant interference up to 100 mg/dL.	Passed (tested up to 146/149 mg/dL).
- Immunoglobulin (IgG)	No interference ≤ 500 mg/L.	Passed (tested up to 1.1 g/L).
- Magnesium	No interference ≤ 75 mmol/L.	Passed (tested up to 75 mmol/L).
- Oxalate	No interference ≤ 2.2 mmol/L.	Passed (tested up to 3.75 mmol/L).
- Phosphate	No interference ≤ 40 mmol/L.	Passed (tested up to 130 mmol/L).
- Urea	No interference ≤ 1000 mmol/L.	Passed (tested up to 1500/1800 mmol/L).
- Uric Acid	No interference ≤ 6 mmol/L.	Passed (tested up to 6 mmol/L).
- Urobilinogen	No interference ≤ 15 mg/dL.	Passed (tested up to 15 mg/dL).
Exogenous Interferences – Drugs	No interference at therapeutic concentrations (except noted).
- Acetaminophen	Not explicitly stated, but results passed up to 3000 mg/L.	No interference up to 3000 mg/L.
- Ascorbic acid	Not explicitly stated, but results passed up to 4000 mg/L.	No interference up to 4000 mg/L.
- Cefoxitin	Not explicitly stated, but results passed up to 12000 mg/L.	No interference up to 12000 mg/L.
- Gentamicin sulfate	Not explicitly stated, but results passed up to 400 mg/L.	No interference up to 400 mg/L.
- Ibuprofen	Not explicitly stated, but results passed up to 500 mg/L.	No interference up to 500 mg/L.
- Levodopa	Not explicitly stated, but results passed up to 1000 mg/L.	No interference up to 1000 mg/L.
- Methyldopa	Not explicitly stated, but results passed up to 2000 mg/L.	No interference up to 2000 mg/L.
- N-Acetylcysteine	Not explicitly stated, but results passed up to 10 mg/L.	No interference up to 10 mg/L.
- Ofloxacine	No interference.	Interference observed (artificially high results). Claim adjusted.
- Phenazopyridine	Not explicitly stated, but results passed up to 50 mg/L.	No interference up to 50 mg/L.
- Salicyluric acid	Not explicitly stated, but results passed up to 100 mg/L.	No interference up to 100 mg/L.
- Tetracycline	Not explicitly stated, but results passed up to 300 mg/L.	No interference up to 300 mg/L.
Method Comparison to Predicate	Predetermined acceptance criteria met. (e.g. slope near 1, intercept near 0, high r)	y = 1.007x + 0.0052, r = 0.995 (Passing Bablok Regression)

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

Precision and Analytical Sensitivity (LoB, LoD, LoQ):
- Precision (Repeatability and Intermediate Precision): 5 human urine sample pools and 2 control samples. Tested for 21 days, 1 run/day, with 2 aliquots per sample in singlicate per part. This setup generates a large number of individual measurements over time to assess variability.
- LoB: One analyte-free sample, measured 10-fold per run across 6 runs (over 4 days) on 3 reagent lots, resulting in 60 measurements per lot.
- LoD: Five human urine samples with low-analyte concentration, measured 2-fold per run across 6 runs (over 4 days) on 3 reagent lots, resulting in 60 measurements per lot.
- LoQ: A low-level sample set prepared by diluting 5 human urine samples, tested in 5 replicates per sample on 4 days, 1 run per day.
Linearity/Assay Reportable Range: Dilution series prepared using human urine sample pools (number of pools not specified, but likely at least one concentrated pool), with 13 concentrations measured on 3 lots in triplicate.
Endogenous Interferences: Two human urine pools (at two transferrin concentrations) were used for each interferent. Each pool was divided into two aliquots (spiked with interferent vs. solvent control). A dilution series of 11 steps was prepared and 3 aliquots per level were tested.
Exogenous Interferences – Drugs: Two human urine sample pools (spiked with approximately 0.433 and 2.48 mg/dL transferrin concentrations) were used for each drug. Each pool was divided into two aliquots (drug spiked vs. solvent control), measured in triplicate.
Method Comparison to Predicate: One hundred and seven (107) routine fresh, never-frozen human urine samples. Two samples were excluded (pH >8, value outside measuring range), so 107 samples were truly used in the comparison.
Data Provenance: The document explicitly states "human urine samples" or "human urine sample pools." For the method comparison, samples were "routine fresh, never-frozen human urine samples." There is no specific mention of the country of origin, but Roche Diagnostics Operations (RDO) is located in Indianapolis, Indiana, USA, and Roche Diagnostics GmbH, Mannheim, Germany is also mentioned as having the establishment registration. The studies are prospective analytical validation studies conducted with collected samples, not retrospective analysis of clinical patient data in the typical sense for imaging.

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

This type of in vitro diagnostic device (IVD) aims for quantitative measurement of a biomarker. Therefore, the "ground truth" is established through analytical reference methods, certified reference materials, or highly rigorous internal validation processes tied to metrological traceability, rather than human expert consensus on subjective findings (as would be the case for an imaging AI).

The ground truth for the test set is established by the analytical measurement on the reference method (for method comparison study), and by the known concentrations/dilutions of samples prepared for analytical studies (e.g., linearity, sensitivity, interference).
No "experts" in the sense of radiologists interpreting images were involved in establishing the ground truth for these analytical performance studies. The accuracy of measurements is verified against the reference standard or predetermined analytical values.

4. Adjudication Method for the Test Set

Not applicable for this type of analytical performance study. Adjudication methods (e.g., 2+1, 3+1) are common in clinical studies where multiple human readers independently assess data (like images) and then a consensus or tie-breaking mechanism is needed to establish ground truth or assess agreement. For an IVD, the "truth" is typically defined by the analytical method itself or a reference method.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, If So, What Was the Effect Size of How Much Human Readers Improve with AI vs Without AI Assistance

Not applicable. This device is a quantitative laboratory assay (a reagent for measuring transferrin in urine), not an AI imaging algorithm that assists human readers. No MRMC study was performed as it is irrelevant to the device's function.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done

Yes, the analytical performance studies (Precision, Analytical Sensitivity, Linearity, Interference, Method Comparison) represent the "standalone" performance of the assay itself (the reagent/instrument system) without human intervention in the result generation beyond operating the analyzer according to instructions. This is the primary form of performance evaluation for an IVD.

7. The Type of Ground Truth Used

For Precision, Analytical Sensitivity, Linearity, and Interference studies: The ground truth is effectively derived from known concentrations in prepared samples (e.g., analyte-free samples, low-concentration samples, dilution series, spiked samples) or reference materials/controls with established values.
For Method Comparison: The ground truth is the measurement obtained from the predicate device, the "N Antisera to Human Transferrin (Siemens) on the BN ProSpec analyzer." This establishes substantial equivalence to an already legally marketed device.

8. The Sample Size for the Training Set

Not applicable. This is not an AI/machine learning device that requires "training data" in the conventional sense. It's a chemical reagent for an established analytical method (immunoturbidimetry). The development process would have involved formulation and optimization, but not "training" on a data set in the way an AI model would be.

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

Not applicable, as there is no "training set" for this type of IVD, which relies on chemical and immunological principles rather than machine learning from data.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

November 5, 2018

Roche Diagnostics Operations (RDO) Lindsay Jones Regulatory Affairs Consultant 9115 Hague Road Indianapolis, Indiana 46250

Re: K182095

Trade/Device Name: Tina-quant Transferrin ver.2 (urine application) Regulation Number: 21 CFR 866.5880 Regulation Name: Transferrin immunological test system Regulatory Class: Class II Product Code: DDG Dated: August 1, 2018 Received: August 3, 2018

Dear Lindsay Jones:

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

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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/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Leonthena R. Carrington -S

Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K182095

Device Name Tina-quant Transferrin ver.2 (urine application)

Indications for Use (Describe)

Tina-quant Transferrin ver.2 (urine application) assay is an in vitro test for the quantitative determination of transferrin in human urine on Roche/Hitachi cobas c systems.

A transferin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the transferrin (an iron-binding and transporting serum protein) in urine. Measurement of transferrin levels aids in the diagnosis of malnutrition, acute inflammation, infection, and red blood cell disorders, such as iron deficiency anemia.

Type of Use (Select one or both, as applicable)
-------------------------------------------------	--

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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Tina-quant Transferrin ver.2 (urine application) 510(k) Summary (K182095)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).

The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the Tina-quant Transferrin ver.2 (urine application).

Submitter Name	Roche Diagnostics Operations (RDO)
Address	9115 Hague RoadIndianapolis, IN 46250, USA
Contact	Lindsay JonesPhone: (317) 521-3544FAX: (317) 521-2324Email: lindsay.jones.lj1 @roche.com
Date Prepared	November 1, 2018
Proprietary Name	Tina-quant Transferrin ver.2 (urine application)
Common Name	Transferrin
Classification Name	Transferrin immunological test system, Class II
Product Codes,Regulation Numbers	DDG, 21 CFR § 866.5880
Predicate Devices	N Antisera to Human Transferrin (K053075)
Establishment Registration	For the Tina-quant Transferrin ver.2 (urine application), the establishmentregistration number for Roche Diagnostics GmbH in Mannheim, Germany is9610126.

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DEVICE DESCRIPTION 1.

Engineering drawings, schematics, and figures are not pertinent to describe the device, as the device is a reagent.

INDICATIONS FOR USE 2.

Tina-quant Transferrin ver.2 (urine application) assay is an in vitro test for the quantitative determination of transferrin in human urine on Roche/Hitachi cobas c systems.

TECHNOLOGICAL CHARACTERISTICS 3.

The Roche transferrin assay is based on the immunological agglutination principle. Human transferrin forms a precipitate with a specific antiserum which is determined turbidimetrically.

The following table compares the Tina-quant Transferrin ver.2 (urine application) with its predicate device, N Antisera to Human Transferrin (K053075). See Table 1 below.

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Feature	Predicate DeviceN Antisera to Human Transferrin(K053075)	Candidate DeviceTina-quant Transferrin ver.2(urine application)
Intended Use	In vitro diagnostic reagents for thequantitative determination of transferrin andhaptoglobin in human serum, heparinizedand EDTA plasma, as well as transferrin inhuman urine by means ofimmunonephelometry on the BN II and BN™ProSpec System.	In vitro test for the quantitativedetermination of transferrin inhuman urine on Roche/Hitachicobas c systems.
Test Principle	Proteins contained in human body fluidsform immune complexes in animmunochemical reaction with specificantibodies. These complexes scatter a beamof light passed through the sample. Theintensity of the scattered light is proportionalto the concentration of the relevant protein inthe sample. The result is evaluated bycomparison with a standard of knownconcentration.	Human transferrin forms aprecipitate with a specificantiserum which is determinedturbidimetrically.
Instrument	BN II/BN ProSpec System	cobas c 501
Reagent Composition	N Antisera are liquid animal sera and areproduced by immunization of rabbits withhighly purified human transferrin. Theconcentration of active antibodies is < 4.2g/L to human transferrin.PreservativeSodium azide <1 g/L	R1: Phosphate buffer: 55mmol/L, pH 7.2; NaCl: 25mmol/L; polyethylene glycol: 5 %;preservativeR2: Anti-human transferrinantibodies (rabbit): dependent ontiter; NaCl: 100 mmol/L;preservative
Sample Type/Matrix	Serum, heparinized and EDTA plasma, urine	Urine
Calibrator	N Protein Standard SL	S1: H2OS2 - S6: Calibrator forautomated systems (C.f.a.s.)Proteins (K133330)
Calibration Interval	The reference curves can be used for aslong as controls with corresponding method-depending target values, e.g. N/T ProteinControls SL/L, M and H or N/T ProteinControl LC, are reproduced within theirrespective range. If a different lot ofantiserum is used, a new reference curvedmust be generated.	Full calibration:• after reagent lot change• as required followingquality controlprocedures
Feature	Predicate DeviceN Antisera to Human Transferrin(K053075)	Candidate DeviceTina-quant Transferrin ver.2(urine application)
Controls	Serum/plasma: N/T Protein Controls SL/L,M, and HUrine: N/T Protein Control LC	PreciControl ClinChem Multi 1 &PreciControl ClinChem Multi 2(K133330)Precinorm Protein & PrecipathProtein (K133330)
Reagent Shelf-Life Stability	Stability at 2 - 8 °C: See expiry date onlabel.Stability once opened: 4 weeks if stored at 2
	– 8 °C securely capped immediately aftereach use and contamination (e.g., bymicroorganisms) is precluded. Duringstorage, N Antisera can develop precipitatesor turbidity which are not caused bymicrobial contamination and which do notaffect their activity. In such cases, theantiserum should be filtered prior to use.Disposable filters with a pore size of 0.45 µmare suitable for this purpose. Do not freeze.	Shelf life at 2 - 8 °C: Seeexpiration date on cobas c packlabel.
Reagent On-Board Stability	A minimum of 5 days at 8 hours per day for 5mL vials, and 3 days at 8 hours per day for 2mL vials or a comparable period of time.	On-board in use and refrigeratedon the analyzer: 8 weeks
Measuring Range	Serum/plasma: 35.0 to 560 mg/dLUrine: 0.22 to 3.5 mg/dL	0.22 to 3.5 mg/dL
Reference Range	Reference interval for urinary transferrin: theconcentration of transferrin in the urine ofhealthy individuals is below the detectionlimit of this method.Nevertheless, each laboratory shoulddetermine its own reference intervals sincevalues may vary depending on the individualpopulation studied.Siemens has not established referenceranges for children.	≤ 1.9 mg/L* (0.19 mg/dL)* The concentration of transferrinin urine of healthy individuals isbelow the detection limit of thismethod.Each laboratory shouldinvestigate the transferability ofthe expected values to its ownpatient population and ifnecessary determine its ownreference ranges.
Feature	Predicate DeviceN Antisera to Human Transferrin(K053075)	Candidate DeviceTina-quant Transferrin ver.2(urine application)
Sample Stability	Suitable samples are human serum,heparinized or EDTA plasma as fresh aspossible (stored no more than 7 days at 2 –8 °C), or stored frozen and fresh humanurine. Serum and plasma samples can bestored at below – 20 °C for up to 3 months ifthey are frozen within 24 hours aftercollection and if repeated freeze thaw cyclesare avoided. Serum samples must becompletely coagulated and, aftercentrifugation, must not contain any particlesor traces of fibrin. Lipemic samples or frozensamples that are turbid after thawing mustbe clarified by centrifugation (10 minutes atapproximately 15,000 x g) prior to testing.Random and timed urine collections aresuitable specimens for testing transferrin inurine. Urine samples must not be frozen.Each urine samples must be centrifugedprior to testing.	4 days at 15 – 25 °C7 days at 2 – 8 °CFreezing and thawing is notallowed

Table 1: Assay Comparison

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NON-CLINICAL PERFORMANCE EVALUATION 4.

The following performance data were provided in support of the substantial equivalence determination:

Precision according to CLSI EP05-A3

Detection Limit: LoB, LoD, LoQ according to CLSI EP17-A2

Linearity according to CLSI EP06-A

Endogenous Interferences

Exogenous Interferences – Drugs

Method Comparison to Predicate

All performance specifications were met.

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4.1. Precision

Repeatability and Intermediate Precision 4.1.1.

Precision experiments were performed in accordance with CLSI Guideline EP05-A3. The testing was run for 21 days, including 1 run per day with 2 parts. Ten dummy samples were included in each part to simulate routine testing in the laboratory. Additionally, 10 dummy samples were included in-between both parts to divide the run into distinct parts. Five human urine sample pools and 2 control samples were tested on one cobas c 501, using 3 lots of reagent. Each sample was tested using 2 aliquots in singlicate per part. Mean, Repeatability (within-run precision) and Intermediate precision (within-lab precision) % CV and SD values were calculated.

Results and Conclusions 4.1.2.

Specimen	Mean (mg/dL)	SD (mg/dL)	CV (%)
PreciControl ClinChemMulti 1	1.98	0.0140	0.7
PreciControl ClinChemMulti 2	3.05	0.0242	0.8
Human Urine 1	0.435	0.00979	2.3
Human Urine 2	0.737	0.00920	1.2
Human Urine 3	1.27	0.0107	0.8
Human Urine 4	2.52	0.0184	0.7
Human Urine 5	3.30	0.0252	0.8

Table 2: Repeatability Summary

Table 3: Intermediate Precision Summary

Specimen	Mean (mg/dL)	SD (mg/dL)	CV (%)
PreciControl ClinChemMulti 1	1.98	0.0158	0.8
PreciControl ClinChemMulti 2	3.05	0.0267	0.9
Human Urine 1	0.435	0.0111	2.5
Human Urine 2	0.737	0.0112	1.5
Human Urine 3	1.23	0.0130	1.1
Human Urine 4	2.52	0.0215	0.9
Human Urine 5	3.30	0.0289	0.9

All data passed the predetermined acceptance criteria.

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Analytical Sensitivity 4.2.

Limit of Blank (LoB) 4.2.1. -

The Limit of Blank (LoB) of the Tina-quant Transferrin ver.2 (urine application) assay on the cobas c 501 was determined according to CLSI EP17-A2. LoB determines the highest measurement result that is likely to be observed for a blank sample with a stated probability. The LoB was determined as the 95th percentile of measurements of blank samples.

One analyte-free sample was measured on 3 lots of reagent in 10-fold determinations per run in 6 runs, distributed over 4 days, on 1 analyzer. In total, 60 measurements were obtained per reagent lot. Data analysis was based on determination of the 95th percentile of the 60 measured values.

LoB Claim: 0.10 mg/dL

Limit of Detection (LoD) 4.2.2.

The Limit of Detection (LoD) of the Tina-quant Transferrin ver.2 (urine application) assay on the cobas c 501 analyzer was determined according to CLSI EP17-A2. The LoD corresponds to the lowest analyte concentration which can be detected (value above the LoB with a probability of 95%).

Five human urine samples with low-analyte concentration (approximately up to 4 times the specified LoB) were measured with 3 lots of reagent in 2-fold determinations per run in 6 runs, distributed over 4 days, on 1 analyzer. In total, 60 measurements were obtained per reagent lot.

LoD Claim: 0.15 mg/dL

Limit of Quantitation (LoQ) 4.2.3.

The Limit of Quantitation (LoQ) of the Tina-quant Transferrin ver.2 (urine application) assay on the cobas c 501 was determined according to CLSI EP17-A2. LoQ determines the lowest amount of analyte in a sample that can be quantitatively determined with stated accuracy and experimental conditions. The LoO was determined as the lowest concentration of analyte which can be quantified with a total error (% CV) of no more than 20%.

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A low level sample set was prepared by diluting 5 human urine samples with analyte-free diluent (water). The low level sample set was tested in 5 replicates per sample on 4 days, 1 run per day, on 1 analyzer.

LoQ Claim: 0.22 mg/dL with 20% CV

Results and Conclusions 4.2.4.

Table 4: LoB, LoD, and LoQ Experimental Determination

	Lot #1 Result(mg/dL)	Lot #2 Result(mg/dL)	Lot #3 Result(mg/dL)	Claim (mg/dL)
Limit of Blank (LoB)	0.0150	0.0130	0.0150	0.10
Limit of Detection(LoD)	0.0382	0.0331	0.0353	0.15
Limit of Quantitation(LoQ)	0.124	0.137	0.143	0.22

All data passed the predetermined acceptance criteria.

Linearity/Assay Reportable Range 4.3.

Regression Analysis 4.3.1. -

The linearity study was conducted to determine whether the measurements across the claimed measuring range for each parameter have a linear relationship. The study was performed according to CLSI EP06-A.

Dilution series were prepared using the human urine sample pools with transferrin concentrations above the upper end of the measuring range. Dilutions were made using water. The dilution series contained 13 concentrations. Sample dilution levels were measured on 3 lots in triplicate on the cobas c 501 analyzer.

Linear regression analysis was done according to EP06-A.

In the first step, a linearity check was performed with 1st order (linear) regression and then with higher order models (quadratic and cubic). A linearity check was performed with a 1* order (linear) regression for reagent lots 1 and 2. A linearity check was performed with a 3th order

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(cubic) regression for reagent lot 3. The linear regression was not forced through the origin. The linear regression was not weighted.

4.3.2. Results and Conclusions

Reagent Lot	Linear Regression Equation(mg/dL)	Pearson CorrelationCoefficient (r)	Measuring Range Claim(mg/dL)
1	y = 1.007x - 0.0189	0.9999	0.22 to 3.5
2	y = 1.008x - 0.0225	0.9999	0.22 to 3.5
3	y = 1.009x - 0.0236	0.9997	0.22 to 3.5

All data passed the predetermined acceptance criteria.

Endogenous Interference Studies 4.4.

The purpose of this study was to evaluate endogenous substances for potential interference with the Tina-quant Transferrin ver.2 (urine application) assay measured on the cobas c 501 analyzer.

The effect on quantitation of analyte in the presence of potential endogenous interfering substances using Tina-quant Transferrin ver.2 (urine application) was determined on the cobas c 501 analyzer at 2 transferrin concentrations and a dilution set of the added interfering substances. Potential interfering substances evaluated include: albumin, citrate, creatinine, glucose, hemoglobin, IgG, magnesium, oxalate, phosphate, urea, uric acid and urobilinogen.

High concentrated stock solutions of the potential interfering substances were prepared in a suitable solvent. Two human urine pools were spiked with defined transferrin concentrations and divided into 2 aliquots. The potential interfering substance was added to 1 aliquot, while the other aliquot was mixed with the same amount of solvent without the potential interfering substance. A dilution series was prepared with 11 dilution steps for each potential interferent by diluting the spiked pool with the dilution pool. Three aliquots per level were tested in 1 run on 1 instrument and 1 lot.

The aliquot containing the potential interfering substance had the same transferrin concentration as the aliquot containing no interfering substance. When diluting those 2 aliquots, the transferrin

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concentration remained constant while the concentration of the potential interferent varied. Thus the effect of increasing concentrations of potential interferent can be determined.

The medians of the measured results were compared to the expected results (aliquot with no potential interfering substance) and the recovery was determined (paired difference testing).

This procedure was repeated for each of the potential interfering substances.

Results and Conclusions 4.4.1.

Potential Interferent	No Interference up to:	Claim
Albumin	Level 1: 5 g/LLevel 2: 5 g/L	No interference ≤ 5000 mg/L
Calcium	Level 1: 9.92 mmol/LLevel 2: 9.80 mmol/L	No interference ≤ 8 mmol/L
Citrate	Level 1: 11 mmol/LLevel 2: 11 mmol/L	No interference ≤ 10 mmol/L
Creatinine	Level 1: 88 mmol/LLevel 2: 88 mmol/L	No interference ≤ 44 mmol/L
Glucose	Level 1: 388 mmol/LLevel 2: 388 mmol/L	No interference ≤ 111 mmol/L
Hemoglobin	Level 1: 146 mg/dLLevel 2: 149 mg/dL	No significant interference up to ahemoglobin concentration of 100mg/dL
Immunoglobulin (IgG)	Level 1: 1.1 g/LLevel 2: 1.1 g/L	No interference ≤ 500 mg/L
Magnesium	Level 1: 75 mmol/LLevel 2: 75 mmol/L	No interference ≤ 75 mmol/L
Oxalate	Level 1: 3.75 mmol/LLevel 2: 3.75 mmol/L	No interference ≤ 2.2 mmol/L
Phosphate	Level 1: 130 mmol/LLevel 2: 130 mmol/L	No interference ≤ 40 mmol/L
Urea	Level 1: 1500 mmol/LLevel 2: 1800 mmol/L	No interference ≤ 1000 mmol/L
Uric Acid	Level 1: 6 mmol/LLevel 2: 6 mmol/L	No interference ≤ 6 mmol/L
Urobilinogen	Level 1: 15 mg/dLLevel 2: 15 mg/dL	No interference ≤ 15 mg/dL

All data passed the predetermined acceptance criteria.

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Exogenous Interferences – Drugs 4.5.

The purpose of this study was to evaluate drugs for potential interference to the Tina-quant Transferrin ver.2 (urine application) assay measured on the cobas c 501 analyzer.

Two human urine sample pools spiked with approximately 0.433 and 2.48 mg/dL transferrin concentrations were divided into 2 aliquots. One aliquot of each concentration was used as the reference sample for transferrin concentration and was not spiked with the drugs, but only the solvent for the drug.

The other aliquots, with either the high or low transferrin concentration, were spiked with the respective amount of drug. Each aliquot containing a defined transferrin concentration and spiked with drug was measured in triplicate in 1 run on 1 analyzer and 1 lot. The defined drug compounds were spiked into samples with concentrations according to EP07-A2 or higher.

Results and Conclusions 4.5.1. -

Potential Drug Interferent	No interference up to:
Acetaminophen	3000 mg/L
Ascorbic acid	4000 mg/L
Cefoxitin	12000 mg/L
Gentamicin sulfate	400 mg/L
Ibuprofen	500 mg/L
Levodopa	1000 mg/L
Methyldopa	2000 mg/L
N-Acetylcysteine	10 mg/L
Ofloxacine	Interference Claim
Phenazopyridine	50 mg/L
Salicyluric acid	100 mg/L
Tetracycline	300 mg/L

Table 7: Potentially Interfering Drugs and Test Concentration Results

All data passed the predetermined acceptance criterion except for ofloxacine.

Claim: Drugs: No interference was found at the therapeutic concentrations using common drug panels. Exceptions: Ofloxacine causes artificially high transferrin results.

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Method Comparison to Predicate 4.6.

A method comparison of the Tina-quant Transferrin ver.2 (urine application) assay on the cobas c 501 versus the predicate device, N Antisera to Human Transferrin (Siemens) on the BN ProSpec analyzer was conducted.

One hundred and seven routine fresh, never-frozen human urine samples were used in the method comparison testing. One of the 109 urine samples collected was removed due to a pH >8 and one was removed due to a measured value outside of the defined measuring range. No patient information was obtained. No samples were spiked or diluted.

The samples were tested in singlicate using the N Antisera to Human Transferrin assay (K053075) from Siemens on the BN ProSpec System and the Tina-quant Transferrin ver.2 (urine application) assay on the cobas c 501 analyzer.

The data was evaluated using Passing Bablok Regression analysis.

4.6.1. Results and Conclusions

Regression analysis results (mg/dL):

y = 1.007x + 0.0052, r = 0.995

Predetermined acceptance criteria for Method Comparison to the Predicate were met.

CLINICAL PERFORMANCE EVALUATION 5.

Not applicable.

ADDITIONAL INFORMATION 6.

Other Devices Required But Not Provided: 6.1.

The Tina-quant Transferrin ver.2 (urine application) continues to use:

Calibrator f. a. s. Proteins (K133330) .
PreciControl ClinChem Multi 1 (K133330) .

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PreciControl ClinChem Multi 2 (K133330) .
Precinorm Protein (K133330) •
Precipath Protein (K133330) •
Diluent NaCl 9% (21 CFR § 864.4010, Class I 510(k) exempt) .

CONCLUSIONS 7.

The submitted information in this premarket notification supports a substantial equivalence decision.

Regulation Number and Section

§ 866.5880 Transferrin immunological test system.

(a)
Identification. A transferrin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the transferrin (an iron-binding and transporting serum protein) in serum, plasma, and other body fluids. Measurement of transferrin levels aids in the diagnosis of malnutrition, acute inflammation, infection, and red blood cell disorders, such as iron deficiency anemia.(b)
Classification. Class II (performance standards).