Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard in vitro diagnostic reagent system for measuring HbA1c using a turbidimetric inhibition immunoassay on a clinical chemistry analyzer. There is no mention of AI or ML in the device description, intended use, or performance studies. The calculations are based on a ratio and user-selected protocols, not complex learning algorithms.

Therapeutic

No
The device is an in vitro diagnostic reagent system used to measure Hemoglobin A1c for the diagnosis and monitoring of diabetes. It does not provide any treatment or therapy.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states, "The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes." This directly indicates its role in diagnosis.

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic reagent system, which includes physical reagents and is used in conjunction with a clinical chemistry analyzer (cobas c 503). It is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use/Indications for Use: The document explicitly states the assay is "an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or venous whole blood". It also mentions its use as "an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes," which are clinical diagnostic purposes.
Device Description: The description details a "reagent system" used to perform a quantitative determination on biological samples (blood). This aligns with the definition of an in vitro diagnostic device.
Performance Studies: The document includes extensive performance studies (Precision, Analytical Sensitivity, Linearity, Interferences, Method Comparison, etc.) which are standard for demonstrating the analytical performance of an IVD.
Predicate Device: The mention of a "Predicate Device(s)" with a K number (K121291) and the name "COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 assay" strongly indicates that this device is being submitted for regulatory review as an IVD, likely through the 510(k) pathway, which requires comparison to a legally marketed predicate IVD.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or venous whole blood on the cobas c 503 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.

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

PDJ, LCP

Device Description

Tina-quant Hemoglobin A1cDx Gen.3 assay is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the cobas c 503 clinical chemistry analyzer.

The assay offers separate applications that are specific to the sample types whole blood and hemolysate. The Whole Blood Application differs from the Hemolysate Application in the hemolyzing step. For the Whole Blood Application, whole blood samples are placed on the analyzer and hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer and hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results. Hemolyzing reagent is part of the test system and is either placed on board the analyzer for the Whole Blood Application or used manually for the Hemolysate Application.

Anticoagulated whole blood is hemolyzed either manually or automatically prior to determination of HbAlc by a turbidimetric inhibition immunoassay. Liberated hemoglobin (Hb) in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum and measured bichromatically. The instrument calculates the % HbAlc from the HbAlc/Hb ratio according to a user selected protocol, either IFCC or NGSP protocols.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

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Intended User / Care Setting

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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

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

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

Precision: Precision measurements were conducted to evaluate repeatability (within-run precision) and intermediate precision (within-laboratory precision) according the CLSI guideline EP5-A3.
Two aliquots per sample were measured once each, in two runs per day, for 21 days, on 3 cobas c 503 analytical units and using 3 reagent lots per system. Ten total samples were evaluated in each run: two controls, PreciControl HbA1c norm and PreciControl HbA1c path, and eight human samples with approximate Hb1Ac concentrations of 4.9%, 6.6%, 7.3%, 8.2%, 12.5%, 14.6%, 12.3% and 13.1% for Whole Blood and 5.0%, 6.6%, 7.3%, 8.3%, 12.5%, 14.7%, 12.1% and 12.9% for Hemolysate.

Analytical Sensitivity - Limit of Blank (LoB): For determination of LoB, one analyte free sample was measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3, in 10-fold determinations. Six runs were distributed over ≥ three days and measured using one cobas c 503 analytical unit. In total, 60 measurements were obtained per lot. Data analysis was based on determination of the 95th percentile of the 60 measured values. LoB was determined according to CLSI guideline EP17-A2.

Analytical Sensitivity - Limit of Detection (LoD): For determination of LoD, five unique human samples with low-analyte concentrations were measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3 in two-fold determinations. The measurements were performed in six runs, over > three days, on one cobas c 503 analytical unit. In total. 60 measurements were obtained per lot. LoD was determined according to CLSI guideline EP17-A2.

Linearity/Assay Reportable Range: Separate dilution series, consisting of at least eleven levels, were prepared for each glycated hemoglobin (HbA1c) and total hemoglobin (Hb) using human hemolysate sample pools. The samples were measured in triplicate and data analysis was performed separately for each sample. The study was performed according to CLSI guideline EP6-A.

Endogenous Interferences: A study evaluated nine endogenous substances for potential interference with measurement of % HbA1c. Pooled whole blood samples, with two hemoglobin A1c levels, one near the medical decision level and one above it, were spiked with the maximum level of the nine interferents, in separate preparations, resulting in eighteen spiked samples which were then hemolyzed with Tina-quant HbA1c Hemolyzing Reagent. Another pool, without interferent, was equally hemolyzed. A > ten-level dilution series was created for each of the eighteen spiked samples, using the interferent-free pools as the diluent. The eighteen dilution series were tested in ten-fold, using one reagent lot, one cobas c 503 analytical unit, in a single run and within one calibration cycle. The mean of the ten replicates was compared to the result from the reference sample (aliquot with no interferent).

Cross-Reactivity: A series of experiments were performed using one reagent lot, on one cobas c 503 analytical unit, in a single run, within one calibration cycle to evaluate potential cross-reactivity with hemoglobin fractions (HbA0, Carbamylated Hb, HbA1(a+b), Acetylated Hb, Labile HbA1c) and glycated albumin. Ten replicates of each sample were analyzed for each dilution level. The median % HbA1c of each dilution level was compared to the median % HbA1c from dilution level zero (without cross-reactant).

Hemoglobin Variants: Hemoglobin variant testing was conducted to determine if significant interference with any of the major hemoglobin variants (HbS, HbC, HbD, HbE, HbA2, Elevated HbF) occurred. Each sample was tested once, in at least one run, on one cobas c 503 analytical unit. Results obtained with the Tina-quant Hemoglobin A1cDx Gen.3 assay were compared to those obtained with the corresponding reference method.

Exogenous Interferences – Drugs: Eighteen commonly used drugs were added to samples at two different HbA1c levels and examined for potential effect on % HbA1c determination. Samples were measured in ten-fold using the cobas c 503 analytical unit. The median value was compared to the reference value (HbA1c sample with no drug added) and the deviation from the reference was calculated.

Sample Matrix Comparison: The study evaluated Hemoglobin A1c determination in the presence of various anticoagulants (K2-EDTA, K3-EDTA, Na Heparin, Li Heparin, NaF/Potassium oxalate, EDTA/Fluoride). At least 40 samples of each anticoagulant and at least 40 half-filled tubes of each anticoagulant were evaluated.

Method Comparison: A method comparison study was performed to compare the sample results from the candidate method, Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit, to results from Tosoh Automated Glycohemoglobin Analyzer HLC-723G8 (Tosoh), the NGSP method. This study was conducted with both the Tina-quant Hemoglobin A1cDx Gen.3 Whole Blood Application and Hemolysate Application. One hundred and seventy-one whole blood samples and one hundred seventy-three hemolysate samples from the secondary NGSP reference laboratory were used. The samples were tested over a 3-day period, with one lot of reagent, on one cobas c 503 analytical unit. The method comparison demonstrated good agreement between Roche Tina-quant Hemoglobin A1cDx Gen.3 assay and the NGSP Tosoh reference method.

Total Error: Total Error (TE) was calculated using the results of bias estimation (%Bias) generated during the method comparison study and precision estimates (%CV) from the precision study, using the equation: %TE = |%Bias| + 1.96 * %CV * (1 + %Bias/100).

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Precision:
- Hemolysate Application, Analytical Unit 1: Intermediate Precision (total) CV ranged from 1.3% to 2.1%.
- Hemolysate Application, Analytical Unit 2: Intermediate Precision (total) CV ranged from 0.9% to 1.7%.
- Hemolysate Application, Analytical Unit 3: Intermediate Precision (total) CV ranged from 1.0% to 1.5%.
- Hemolysate Application, 3 Lots and 3 Analytical Units: Reproducibility (total) CV ranged from 1.1% to 1.9%.
- Whole Blood Application, Analytical Unit 1: Intermediate Precision (total) CV ranged from 1.2% to 1.5%.
- Whole Blood Application, Analytical Unit 2: Intermediate Precision (total) CV ranged from 1.1% to 1.6%.
- Whole Blood Application, Analytical Unit 3: Intermediate Precision (total) CV ranged from 1.2% to 1.6%.
- Whole Blood Application, 3 Lots and 3 Analytical Units: Reproducibility (total) CV ranged from 1.3% to 1.6%.
Analytical Sensitivity (LoB):
- Hb LoB: 0.0530 mmol/L (0.085 g/dL)
- HbA1c LoB: 0.0220 mmol/L (0.035 g/dL)
- Corresponds to 15 mmol/mol (IFCC) and 3.5 % HbA1c (DCCT/NGSP) at 13.2 g/dL Hb.
Analytical Sensitivity (LoD):
- Hb LoD: 0.119 mmol/L (0.192 g/dL)
- HbA1c LoD: 0.0437 mmol/L (0.07 g/dL)
- Corresponds to 22 mmol/mol (IFCC) and 4.2 % HbA1c (DCCT/NGSP) at 13.2 g/dL Hb.
Linearity/Assay Reportable Range:
- Hemolysate Hb: 3.04 g/dL - 40.4 g/dL
- Hemolysate HbA1c: 0.293 g/dL - 2.87 g/dL
- Claimed measuring range: 23-196 mmol/mol HbA1c (IFCC) and 4.2-20.1 % HbA1c (DCCT/NGSP).
Endogenous Interferences (claimed maximum concentration without interference):
- Bilirubin: 60 mg/dL
- Ditaurobilirubin: 60 mg/dL
- Lipemia: 400 mg/dL
- Rheumatoid Factors: 750 IU/mL
- Total Protein: 21 g/dL
- Albumin: 60 g/L
- Immunoglobulin (IgG): 60 g/L
- Glucose: 1000 mg/dL
- Triglycerides: 1584 mg/dL
Cross-Reactivity (Max Whole Blood Cross-Reactant Concentration with no Interference):
- HbA0: 120 g/dL
- HbA1(a+b): 0.96 g/dL (HbA1c Level 1), 1.6 g/dL (HbA1c Level 2)
- Carbamylated Hb: 2.0 g/dL
- Acetylated Hb: 2.0 g/dL
- Glycated Albumin: 10 g/dL
- Labile HbA1c: 1000 mg/dL
Hemoglobin Variant Testing (Percent Relative Bias from Reference Method):
- HbS: -2.5% (around 6.5% HbA1c), -4.0% (around 9% HbA1c)
- HbC: -3.9% (around 6.5% HbA1c), -6.0% (around 9% HbA1c)
- HbE: -0.1% (around 6.5% HbA1c), -1.2% (around 9% HbA1c)
- HbD: -1.8% (around 6.5% HbA1c), -2.6% (around 9% HbA1c)
- HbA2: -1.0% (around 6.5% HbA1c), 0.4% (around 9% HbA1c)
- Elevated HbF: Specimens containing high amounts of HbF (>7%) may yield lower than expected HbA1c values.
Method Comparison:
- Whole Blood Application: Mean bias vs. NGSP TOSOH: -0.046%. Mean bias at lower 95% CI: -0.410%, upper 95% CI: 0.318%.
- Hemolysate Application: Mean bias vs. NGSP TOSOH: 0.046%. Mean bias at lower 95% CI: -0.338%, upper 95% CI: 0.431%.
Total Error:
- Hemolysate Application: Range from 3.0% to 4.4%.
- Whole Blood Application: Range from 3.1% to 5.2%.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 assay, K121291

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 862.1373 Hemoglobin A1c test system.

(a)
Identification. A hemoglobin A1c test system is a device used to measure the percentage concentration of hemoglobin A1c in blood. Measurement of hemoglobin A1c is used as an aid in the diagnosis of diabetes mellitus and as an aid in the identification of patients at risk for developing diabetes mellitus.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The device must have initial and annual standardization verification by a certifying glycohemoglobin standardization organization deemed acceptable by FDA.
(2) The premarket notification submission must include performance testing to evaluate precision, accuracy, linearity, and interference, including the following:
(i) Performance testing of device precision must, at a minimum, use blood samples with concentrations near 5.0 percent, 6.5 percent, 8.0 percent, and 12 percent hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least three lots of the device and three instruments, as applicable.
(ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the device and compare results of the new device to results of a standardized test method. Results must demonstrate little or no bias versus the standardized method.
(iii) Total error of the new device must be evaluated using single measurements by the new device compared to results of the standardized test method, and this evaluation must demonstrate a total error less than or equal to 6 percent.
(iv) Performance testing must demonstrate that there is little to no interference from common hemoglobin variants, including Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2, and Hemoglobin S.
(3) When assay interference from Hemoglobin F or interference with other hemoglobin variants with low frequency in the population is observed, a warning statement must be placed in a black box and must appear in all labeling material for these devices describing the interference and any affected populations.

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.

March 26, 2020

Roche Diagnostics Operations Inc. Leslie Patterson Regulatory Affairs Principal 9115 Hague Rd Indianapolis, Indiana 46250

Re: K193053

Trade/Device Name: Tina-quant Hemoglobin A1cDx Gen.3 Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin A1c Test System Regulatory Class: Class II Product Code: PDJ, LCP Dated: February 13, 2020 Received: February 14, 2020

Dear Leslie Patterson:

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

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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,

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

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Indications for Use

510(k) Number (if known) K193053

Device Name Tina-quant Hemoglobin A1cDx Gen.3

Indications for Use (Describe)

The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or venous whole blood on the cobas c 503 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.

Type of Use (Select one or both, as applicable)
☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)

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Tina-quant Hemoglobin A1cDx Gen.3 510(k) Summary (K193053)

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 Hemoglobin A1cDx Gen.3 assay.

Submitter Name	Roche Diagnostics Operations Inc.
Address	9115 Hague Road
P.O. Box 50416
Indianapolis, IN 46250-0457
Contact	Leslie Patterson
Phone: (317) 521-7307
FAX: (317) 521-2324
Email: leslie.patterson@roche.com
Date Prepared	February 13, 2020
Proprietary Name	Tina-quant Hemoglobin A1cDx Gen.3
Common Name	Glycosylated Hemoglobin Assay
Classification Name	Hemoglobin A1c test system
Product Codes,
Regulation Numbers	PDJ, 862.1373
LCP, 864.7470
Predicate Device	COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 assay, K121291
Establishment Registration	1823260, Roche Diagnostics Corporation

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1. DEVICE DESCRIPTION

Tina-quant Hemoglobin A1cDx Gen.3 assay is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the cobas c 503 clinical chemistry analyzer.

The assay offers separate applications that are specific to the sample types whole blood and hemolysate. The Whole Blood Application differs from the Hemolysate Application in the hemolyzing step. For the Whole Blood Application, whole blood samples are placed on the analyzer and hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer and hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results. Hemolyzing reagent is part of the test system and is either placed on board the analyzer for the Whole Blood Application or used manually for the Hemolysate Application.

Anticoagulated whole blood is hemolyzed either manually or automatically prior to determination of HbAlc by a turbidimetric inhibition immunoassay. Liberated hemoglobin (Hb) in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum and measured bichromatically. The instrument calculates the % HbAlc from the HbAlc/Hb ratio according to a user selected protocol, either IFCC or NGSP protocols.

Test Principle 1.1.

This method uses tetradecyltrimethylammonium bromide (TTAB) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to remove labile HbA1c is not necessary. All hemoglobin variants which are glycated at the ß -chain N-terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of patients having uremia or the most frequent hemoglobinopathies (HbAC, HbAC, HbAD) can be determined using this assay.

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1.2. Hemoglobin A1c

The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.

. Sample and addition of R1 (buffer/antibody):
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, formation of insoluble complexes does not take place.
Addition of R3 (buffer/polyhapten) and start of reaction: .
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibodypolyhapten complex which can be determined turbidimetrically.

1.3. Hemoglobin

Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently, not necessary.

Final HbA1c Result 1.4.

The final result is expressed as mmol/mol HbA1c and is calculated from the HbA1c/Hb ratio as follows:

Protocol 1 (mmol/mol HbA1c acc. to IFCC): HbA1c (mmol/mol) = (HbA1c/Hb) × 1000

Protocol 2 (% HbA1c acc. to DCCT/NGSP): HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15

1.5. Standardization

Traceability: This method has been standardized against the approved IFCC reference method for the measurement of HbA1c in human blood and can be transferred to results traceable to DCCT/NGSP by calculation.

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2. INDICATIONS FOR USE

The Tina-quant Hemoglobin A1cDx Gen.3 assay is intended for use as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. It is an in vitro diagnostics reagent system intended for quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or venous whole blood on the cobas c 503 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.

TECHNOLOGICAL CHARACTERISTICS 3.

The following table compares the Tina-quant Hemoglobin A1cDx Gen.3 assay with its predicate device, COBAS INTEGRA 800 Tina-quant Hemoglobin A1cDx Gen.2 (K121291).

| Feature | Predicate Device:
COBAS INTEGRA 800 Tina-quant
Hemoglobin A1cDx Gen.2 K121291 | Submitted Device:
Tina-quant Hemoglobin A1cDx Gen.3 |
|-----------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | This test is to be used as an aid in diagnosis of
diabetes and as an aid in identifying patients
who may be at risk for developing diabetes. It
is an in vitro diagnostic reagent system
intended for quantitative determination of
mmol/mol hemoglobin A1c (IFCC) and %
hemoglobin A1c (DCCT/NGSP) in hemolysate
or venous whole blood. HbA1c determinations
are useful for monitoring of long-term blood
glucose control in individuals with diabetes
mellitus. | Same |
| Sample Types | Anticoagulated venous blood
Acceptable anticoagulants for both the
hemolysate and whole blood applications
include:
• Li-Heparin
• K2-EDTA
• K3-EDTA
• Fluoride/potassium oxalate
• Na-Heparin
• NaF/Na2-EDTA | Anticoagulated venous blood
Acceptable anticoagulants for both the
hemolysate and whole blood applications
include:
• Li-Heparin
• K2-EDTA
• K3-EDTA
• Fluoride/potassium oxalate
• Na-Heparin
• EDTA Fluoride |
| Instrument
Platform | COBAS Integra 800
• Absorbance Photometry | cobas c 503 |
| Calibrator | Cfas HbA1c | Same |
| Feature | Predicate Device:
COBAS INTEGRA 800 Tina-quant
Hemoglobin A1cDx Gen.2 K121291 | Submitted Device:
Tina-quant Hemoglobin A1cDx Gen.3 |
| Calibration
Frequency | • After 29 days on-board the analyzer
• After reagent lot change
• As required following quality control
procedures | Same |
| Calibration Mode | Logit/Log 5 | Hb: linear
HbA1c : RCM4 |
| Controls | PreciControl HbA1c norm
PreciControl HbA1c path | Same |
| Reagent Stability | Unopened:
• 2-8 °C until expiration date
On-board in use:
• 2-8 °C for 28 days | Same |
| Reporting Units | • % HbA1c (NGSP/DCCT)
• mmol/mol (IFCC) | Same |
| Antibody | Polyclonal anti-HbA1c from sheep blood | Same |
| Test Principle | The anticoagulated whole blood specimen is
hemolyzed automatically on the COBAS
INTEGRA 800 analyzers with COBAS
INTEGRA Hemolyzing Reagent Gen.2. This
method uses TTAB as the detergent in the
hemolyzing reagent to eliminate interference
from leukocytes (TTAB does not lyse
leukocytes). Sample pretreatment to remove
labile HbA1c is not necessary. All hemoglobin
variants which are glycated at the β-chain N-
terminus and which have antibody-recognizable
regions identical to that of HbA1c are
determined by this assay. Consequently, the
metabolic state of diabetic patients having
uremia or the most frequent
hemoglobinopathies (HbAS, HbAC, HbAE,
HbAD) can be determined by this assay. | This method uses
tetradecyltrimethylammonium bromide (TTAB)
as the detergent in the hemolyzing reagent to
eliminate interference from leukocytes (TTAB
does not lyse leukocytes). Sample
pretreatment to remove labile HbA1c is not
necessary. All hemoglobin variants which are
glycated at the β -chain N-terminus and which
have antibody-recognizable regions identical to
that of HbA1c are determined by this assay.
Consequently, the metabolic state of patients
having uremia or the most frequent
hemoglobinopathies (HbAS, HbAC, HbAE,
HbAD) can be determined using this assay. |
| Determination of
HbA1c | Turbidimetric immunoinhibition (TINIA).
Antigen-antibody complexes are formed
and excess Ab aggregate with polyhapten to
form insoluble complexes | HbA1c determination is based on the
turbidimetric inhibition immunoassay (TINIA)
for hemolyzed whole blood. Glycohemoglobin
in the sample reacts with anti-HbA1c antibody
to form soluble antigen-antibody complexes.
Polyhaptens react with excess anti-HbA1c
antibodies to form an insoluble antibody-
polyhapten complex which can be measured
turbidimetrically. |
| Determination of
Hb | Bichromatic photometric determination
after conversion to a colored derivate | Liberated hemoglobin in the hemolyzed
sample is converted to a derivative having a
characteristic absorption spectrum which is
measured bichromatically. |
| Feature | Predicate Device:
COBAS INTEGRA 800 Tina-quant
Hemoglobin A1cDx Gen.2 K121291 | Submitted Device:
Tina-quant Hemoglobin A1cDx Gen.3 |
| Determination
of % HbA1c | The final result is expressed as % HbA1c and
is calculated from the HbA1c/Hb ratio per
DCCT/NGSP as follows:
$HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15$ | Same |
| Measuring Range | Hemoglobin: 4-35 g/dL
HbA1c: 0.3-3.4 g/dL
This corresponds to a measuring range of
23-258 mmol/mol HbA1c (IFCC)
and 4.3-24.8 % HbA1c (DCCT/NGSP) at a
typical hemoglobin concentration of 13.2 g/dL. | Hemoglobin: 4-40 g/dL (2.48-24.8 mmol/L)
HbA1c: 0.3-2.6 g/dL (0.186-1.61 mmol/L)
This corresponds to a measuring range of 23-
196 mmol/molHbA1c (IFCC) and 4.2-20.1 %
HbA1c (DCCT/NGSP) at a typical hemoglobin
concentration of 13.2 g/dL (8.2 mmol/L). |
| Traceability | The assigned HbA1c and total hemoglobin
values of the cobas c Tina-quant Hemoglobin
A1cDx Gen.3 assay is certified with the
National Glycohemoglobin Standardization
Program (NGSP). NGSP certification is
repeated annually. | Same |
| Reagent
Composition | R1 Antibody Reagent:
• MES (2-morpholinoethane sulfonic acid)
buffer: 0.025 mol/L
• TRIS (Tris(hydroxymethyl) aminomethane)
buffer: 0.015 mol/L, pH 6.2
• HbA1c antibody (ovine serum): ≥ 0.5 mg/ml
• detergents; stabilizers; preservatives
SR Polyhapten Reagent:
• MES buffer: 0.025 mol/L
• TRIS buffer: 0.015 mol/L, pH 6.2
• HbA1c polyhapten: > 8µg/mL
• stabilizers; preservatives
A1CD (Hemolyzing Reagent):
• Aqueous buffered matrix, pH 7.25
• Tetradecyltrimethylammonium bromide: 36
g/L
• sodium dihydrogenphosphate monohydrate:
16 mmol/L
• sodium monohydrogenphosphate dihydrate:
64 mmol/L
stabilizers; preservatives | R1 Antibody Reagent:
•MES (2-morpholinoethane sulfonic
acid) buffer: 0.025 mol/L
•TRIS (Tris(hydroxymethyl)aminomethane)
buffer: 0.015 mol/L, pH 6.2
•HbA1c antibody (ovine serum): ≥ 0.5 mg/ml
• detergents; stabilizers; preservatives
R3 Polyhapten Reagent:
•MES buffer: 0.025 mol/L
•TRIS buffer: 0.015 mol/L, pH 6.2
•HbA1c polyhapten: > 8 µg/mL
• detergents; stabilizers; preservatives
A1CD (Hemolyzing Reagent):
• Aqueous buffered matrix, pH 7.25
• Tetradecyltrimethylammonium bromide: 36
g/L
• Sodium dihydrogenphosphate monohydrate:
16 mmol/L
• Sodium monohydrogenphosphate dihydrate:
64 mmol/L
stabilizers; preservatives |

Table 1: Substantial Equivalence Assay Comparison

7

8

9

4. NON-CLINICAL PERFORMANCE EVALUATION

Performance characteristics were evaluated with Tina-quant Hemoglobin A1cDx Gen.3 on the cobas c 503 analytical unit.

Tina-quant Hemoglobin A1cDx Gen.3 offers two sample type specific applications, Hemolysate Application and Whole Blood Application, one for manually hemolyzed samples and one for whole blood samples respectively.

The Tina-quant Hemoglobin A1cDx Gen.3 assay first measures total hemoglobin (Hb) and glycated hemoglobin (HbA1c) in terms of either g/dL or mmol/L. Then the analyzer calculates the HbA1c/Hb ratio according to either IFCC or DCCT/NGSP. IFCC protocol reports the ratio in terms of mmol/mol HbA1c while the DCCT/NGSP protocol reports the ratio in terms of % HbA1c. Performance characteristics that support the measuring ranges claimed for Hb and HbA1c included limit of detection and linearity. These results were reported in terms of Hb and HbA1c individually. Patient sample values are reported in terms of the ratio of glycated to total hemoglobin. Method comparison, control recovery and precision were evaluated in terms of the ratio.

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

Precision 4.1.

Repeatability and Intermediate Precision 4.1.1.

Precision measurements were conducted to evaluate repeatability (within-run precision) and intermediate precision (within-laboratory precision) according the CLSI guideline EP5-A3.

Two aliquots per sample were measured once each, in two runs per day, for 21 days, on 3 cobas c 503 analytical units and using 3 reagent lots per system. Ten total samples were evaluated in each run: two controls, PreciControl HbA1c norm and PreciControl HbA1c path, and eight human samples with approximate Hb1Ac concentrations of 4.9%, 6.6%, 7.3%, 8.2%, 12.5%, 14.6%, 12.3% and 13.1% for Whole Blood and 5.0%, 6.6%, 7.3%, 8.3%, 12.5%, 14.7%, 12.1% and 12.9% for Hemolysate.

10

The samples were randomized within each run. For each sample, the following was calculated: mean, repeatability and intermediate precision as CV and SD values and the upper 95% confidence interval for SD and CV values.

| Mean
% HbA1c | Repeatability (error) | | Between-Run | | Between-Day | | Between-lot | | Intermediate Precision
(total) | |
|-----------------|-----------------------|--------|-------------|--------|-------------|--------|-------------|--------|-----------------------------------|--------|
| | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Hem 1
4.96 | 0.025 | 0.5 | 0.006 | 0.1 | 0.071 | 1.4 | 0.016 | 0.3 | 0.077 | 1.6 |
| Hem 2
6.62 | 0.027 | 0.4 | 0.013 | 0.2 | 0.053 | 0.8 | 0.059 | 0.9 | 0.085 | 1.3 |
| Hem 3
7.32 | 0.035 | 0.5 | 0.000 | 0.0 | 0.053 | 0.7 | 0.067 | 0.9 | 0.092 | 1.3 |
| Hem 4
8.32 | 0.039 | 0.5 | 0.009 | 0.1 | 0.056 | 0.7 | 0.083 | 1.0 | 0.108 | 1.3 |
| Hem 5
12.54 | 0.057 | 0.5 | 0.011 | 0.1 | 0.100 | 0.8 | 0.203 | 1.6 | 0.234 | 1.9 |
| HE 006
14.77 | 0.077 | 0.5 | 0.013 | 0.1 | 0.148 | 1.0 | 0.268 | 1.8 | 0.316 | 2.1 |
| HE_007
12.14 | 0.055 | 0.5 | 0.023 | 0.2 | 0.100 | 0.8 | 0.181 | 1.5 | 0.215 | 1.8 |
| HE_008
12.94 | 0.072 | 0.6 | 0.000 | 0.0 | 0.111 | 0.9 | 0.188 | 1.5 | 0.230 | 1.8 |
| PCA1N
5.53 | 0.024 | 0.4 | 0.009 | 0.2 | 0.059 | 1.1 | 0.028 | 0.5 | 0.071 | 1.3 |
| PCA1P
10.89 | 0.055 | 0.5 | 0.026 | 0.2 | 0.085 | 0.8 | 0.146 | 1.3 | 0.179 | 1.6 |

Table 2: Precision Results – Hemolysate Application, Analytical Unit 1

Table 3: Precision Results - Hemolysate Application, Analytical Unit 2

| Mean
% HbA1c | | Repeatability (error) | Between-Run | | Between-Day | | | Between-lot | Intermediate Precision
(total) | |
|-----------------|-------|-----------------------|-------------|--------|-------------|--------|-------|-------------|-----------------------------------|--------|
| | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Hem 1
4.96 | 0.027 | 0.5 | 0.005 | 0.1 | 0.034 | 0.7 | 0.015 | 0.3 | 0.046 | 0.9 |
| Hem 2
6.59 | 0.035 | 0.5 | 0.000 | 0.0 | 0.038 | 0.6 | 0.057 | 0.9 | 0.077 | 1.2 |
| Hem 3
7.29 | 0.041 | 0.6 | 0.000 | 0.0 | 0.043 | 0.6 | 0.068 | 0.9 | 0.090 | 1.2 |
| Hem 4
8.28 | 0.039 | 0.5 | 0.015 | 0.2 | 0.046 | 0.6 | 0.093 | 1.1 | 0.112 | 1.4 |
| Hem 5
12.43 | 0.069 | 0.6 | 0.027 | 0.2 | 0.038 | 0.3 | 0.175 | 1.4 | 0.193 | 1.6 |
| HE 006
14.68 | 0.085 | 0.6 | 0.011 | 0.1 | 0.060 | 0.4 | 0.220 | 1.5 | 0.243 | 1.7 |
| HE_007
12.05 | 0.063 | 0.5 | 0.018 | 0.1 | 0.036 | 0.3 | 0.163 | 1.4 | 0.179 | 1.5 |
| HE_008
12.85 | 0.071 | 0.6 | 0.034 | 0.3 | 0.053 | 0.4 | 0.177 | 1.4 | 0.201 | 1.6 |
| PCA1N
5.52 | 0.030 | 0.5 | 0.008 | 0.1 | 0.029 | 0.5 | 0.024 | 0.4 | 0.049 | 0.9 |
| PCA1P
10.81 | 0.074 | 0.7 | 0.000 | 0.0 | 0.041 | 0.4 | 0.134 | 1.2 | 0.159 | 1.5 |

Table 4: Precision Results – Hemolysate Application, Analytical Unit 3

| Mean %
HbA1c | Repeatability (error)
SD | Repeatability (error)
CV (%) | Between-Run
SD | Between-Run
CV (%) | Between-Day
SD | Between-Day
CV (%) | Between-lot
SD | Between-lot
CV (%) | Intermediate Precision (total)
SD | Intermediate Precision (total)
CV (%) |
|-----------------|-----------------------------|---------------------------------|-------------------|-----------------------|-------------------|-----------------------|-------------------|-----------------------|--------------------------------------|------------------------------------------|
| Hem 1
4.94 | 0.027 | 0.5 | 0.010 | 0.2 | 0.031 | 0.6 | 0.023 | 0.5 | 0.048 | 1.0 |
| Hem 2
6.57 | 0.030 | 0.5 | 0.004 | 0.1 | 0.035 | 0.5 | 0.049 | 0.7 | 0.067 | 1.0 |

11

| Hem 3

7.28	0.032	0.4	0.000	0.0	0.038	0.5	0.054	0.7	0.073	1.0
Hem 4
8.28	0.040	0.5	0.000	0.0	0.041	0.5	0.070	0.8	0.091	1.1
Hem 5
12.43	0.063	0.5	0.024	0.2	0.045	0.4	0.162	1.3	0.181	1.5
HE 006
14.68	0.075	0.5	0.000	0.0	0.050	0.3	0.240	1.6	0.256	1.7
HE 007
12.07	0.064	0.5	0.000	0.0	0.039	0.3	0.146	1.2	0.164	1.4
HE_008
12.84	0.072	0.6	0.000	0.0	0.052	0.4	0.150	1.2	0.174	1.4
PCA1N
5.49	0.026	0.5	0.008	0.2	0.035	0.6	0.024	0.4	0.050	0.9
PCA1P
10.78	0.071	0.7	0.000	0.0	0.048	0.4	0.116	1.1	0.144	1.3

Table 5: Precision Results - Hemolysate Application, 3 Lots and 3 Analytical Units
------------------------------------------------------------------------------------	--

| Mean,
%HbA1c | | Repeatability
(error) | | Between-Run | | Between-Day | | Between-Lot | | Between-Device | | Reproducibility
(total) | |
|------------------|-------|--------------------------|-------|-------------|-------|-------------|-------|-------------|-------|----------------|-------|----------------------------|--|
| | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | |
| Hem 1,
4.96 | 0.026 | 0.5 | 0.007 | 0.1 | 0.049 | 1.0 | 0.019 | 0.4 | 0.009 | 0.2 | 0.059 | 1.2 | |
| Hem 2,
6.59 | 0.031 | 0.5 | 0.006 | 0.1 | 0.042 | 0.6 | 0.055 | 0.8 | 0.023 | 0.3 | 0.080 | 1.2 | |
| Hem 3,
7.30 | 0.036 | 0.5 | 0.000 | 0.0 | 0.045 | 0.6 | 0.063 | 0.9 | 0.019 | 0.3 | 0.088 | 1.2 | |
| Hem 4,
8.29 | 0.039 | 0.5 | 0.005 | 0.1 | 0.049 | 0.6 | 0.082 | 1.0 | 0.019 | 0.2 | 0.105 | 1.3 | |
| Hem 5,
12.47 | 0.063 | 0.5 | 0.022 | 0.2 | 0.070 | 0.6 | 0.179 | 1.4 | 0.061 | 0.5 | 0.212 | 1.7 | |
| HE_006,
14.71 | 0.079 | 0.5 | 0.010 | 0.1 | 0.098 | 0.7 | 0.242 | 1.6 | 0.053 | 0.4 | 0.278 | 1.9 | |
| HE_007,
12.08 | 0.061 | 0.5 | 0.016 | 0.1 | 0.067 | 0.6 | 0.163 | 1.3 | 0.048 | 0.4 | 0.193 | 1.6 | |
| HE_008,
12.88 | 0.072 | 0.6 | 0.017 | 0.1 | 0.078 | 0.6 | 0.172 | 1.3 | 0.055 | 0.4 | 0.210 | 1.6 | |
| PCA1N,
5.51 | 0.027 | 0.5 | 0.008 | 0.2 | 0.043 | 0.8 | 0.027 | 0.5 | 0.022 | 0.4 | 0.062 | 1.1 | |
| PCA1P,
10.83 | 0.067 | 0.6 | 0.000 | 0.0 | 0.062 | 0.6 | 0.132 | 1.2 | 0.054 | 0.5 | 0.169 | 1.6 | |

Table 6: Precision Results – Whole Blood Application, Analytical Unit 1

| Mean | Repeatability (error) | | Between-Run | | Between-Day | | Between-Lot | | Intermediate Precision
(total) | |
|------------------|-----------------------|-------|-------------|-------|-------------|-------|-------------|-------|-----------------------------------|-------|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| WB 1,
4.87 | 0.038 | 0.8 | 0.000 | 0.0 | 0.043 | 0.9 | 0.045 | 0.9 | 0.073 | 1.5 |
| WB 2,
6.60 | 0.026 | 0.4 | 0.015 | 0.2 | 0.030 | 0.4 | 0.071 | 1.1 | 0.083 | 1.3 |
| WB 3,
7.37 | 0.032 | 0.4 | 0.018 | 0.2 | 0.032 | 0.4 | 0.084 | 1.1 | 0.097 | 1.3 |
| WB 4,
8.24 | 0.039 | 0.5 | 0.008 | 0.1 | 0.048 | 0.6 | 0.089 | 1.1 | 0.108 | 1.3 |
| WB 5,
12.59 | 0.056 | 0.4 | 0.025 | 0.2 | 0.061 | 0.5 | 0.154 | 1.2 | 0.177 | 1.4 |
| WB_006,
14.69 | 0.079 | 0.5 | 0.043 | 0.3 | 0.067 | 0.5 | 0.168 | 1.1 | 0.202 | 1.4 |
| WB_007,
12.34 | 0.062 | 0.5 | 0.032 | 0.3 | 0.057 | 0.5 | 0.152 | 1.2 | 0.176 | 1.4 |
| WB_008,
13.14 | 0.061 | 0.5 | 0.015 | 0.1 | 0.055 | 0.4 | 0.165 | 1.3 | 0.185 | 1.4 |
| PCA1N,
5.51 | 0.029 | 0.5 | 0.002 | 0.0 | 0.039 | 0.7 | 0.044 | 0.8 | 0.066 | 1.2 |
| PCA1P,
11.20 | 0.055 | 0.5 | 0.011 | 0.1 | 0.057 | 0.5 | 0.129 | 1.2 | 0.152 | 1.4 |

12

| Mean
%HbA1c | Repeatability (error) | | Between-Run | | Between-Day | | Between-Lot | | Intermediate Precision
(total) | |
|------------------|-----------------------|-------|-------------|-------|-------------|-------|-------------|-------|-----------------------------------|-------|
| | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| WB 1,
4.88 | 0.032 | 0.7 | 0.013 | 0.3 | 0.026 | 0.5 | 0.031 | 0.6 | 0.053 | 1.1 |
| WB 2,
6.58 | 0.031 | 0.5 | 0.010 | 0.2 | 0.025 | 0.4 | 0.077 | 1.2 | 0.087 | 1.3 |
| WB 3,
7.35 | 0.040 | 0.5 | 0.000 | 0.0 | 0.029 | 0.4 | 0.091 | 1.2 | 0.103 | 1.4 |
| WB 4,
8.21 | 0.042 | 0.5 | 0.011 | 0.1 | 0.035 | 0.4 | 0.106 | 1.3 | 0.120 | 1.5 |
| WB 5,
12.53 | 0.070 | 0.6 | 0.029 | 0.2 | 0.047 | 0.4 | 0.164 | 1.3 | 0.186 | 1.5 |
| WB_006,
14.62 | 0.095 | 0.6 | 0.035 | 0.2 | 0.091 | 0.6 | 0.198 | 1.4 | 0.240 | 1.6 |
| WB_007,
12.25 | 0.078 | 0.6 | 0.000 | 0.0 | 0.035 | 0.3 | 0.176 | 1.4 | 0.195 | 1.6 |
| WB_008,
13.06 | 0.073 | 0.6 | 0.009 | 0.1 | 0.052 | 0.4 | 0.163 | 1.2 | 0.186 | 1.4 |
| PCA1N,
5.51 | 0.036 | 0.6 | 0.000 | 0.0 | 0.028 | 0.5 | 0.044 | 0.8 | 0.063 | 1.1 |
| PCA1P,
11.13 | 0.062 | 0.6 | 0.033 | 0.3 | 0.043 | 0.4 | 0.162 | 1.5 | 0.182 | 1.6 |

Table 7: Precision Results – Whole Blood Application, Analytical Unit 2

					Table 8: Precision Results - Whole Blood Application, Analytical Unit 3
--	--	--	--	--	--------------------------------------------------------------------------	--	--

| Mean | Repeatability (error) | | Between-Run | | Between-Day | | Between-Lot | | Intermediate Precision
(total) | |
|------------------|-----------------------|-------|-------------|-------|-------------|-------|-------------|-------|-----------------------------------|-------|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| WB 1,
4.86 | 0.029 | 0.6 | 0 | 0.0 | 0.035 | 0.7 | 0.044 | 0.9 | 0.064 | 1.3 |
| WB 2,
6.55 | 0.031 | 0.5 | 0.009 | 0.1 | 0.025 | 0.4 | 0.080 | 1.2 | 0.090 | 1.4 |
| WB 3,
7.31 | 0.039 | 0.5 | 0.006 | 0.1 | 0.026 | 0.4 | 0.090 | 1.2 | 0.101 | 1.4 |
| WB 4,
8.17 | 0.043 | 0.5 | 0.012 | 0.2 | 0.036 | 0.4 | 0.096 | 1.2 | 0.111 | 1.4 |
| WB 5,
12.51 | 0.079 | 0.6 | 0.000 | 0.0 | 0.064 | 0.5 | 0.154 | 1.2 | 0.184 | 1.5 |
| WB_006,
14.62 | 0.082 | 0.6 | 0.000 | 0.0 | 0.107 | 0.7 | 0.191 | 1.3 | 0.234 | 1.6 |
| WB_007,
12.23 | 0.068 | 0.6 | 0.028 | 0.2 | 0.056 | 0.5 | 0.157 | 1.3 | 0.182 | 1.5 |
| WB_008,
13.05 | 0.083 | 0.6 | 0.000 | 0.0 | 0.064 | 0.5 | 0.162 | 1.2 | 0.193 | 1.5 |
| PCA1N,
5.50 | 0.030 | 0.6 | 0.006 | 0.1 | 0.028 | 0.5 | 0.051 | 0.9 | 0.066 | 1.2 |
| PCA1P,
11.10 | 0.071 | 0.6 | 0.000 | 0.0 | 0.043 | 0.4 | 0.137 | 1.2 | 0.160 | 1.4 |

13

| Mean | | Repeatability
(error) | | Between-Run | | Between-Day | | Between-Lot | | Between-Device | | Reproducibility
(total) | |
|------------------|-------|--------------------------|-------|-------------|-------|-------------|-------|-------------|-------|----------------|-------|----------------------------|--|
| %HbA1c | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | |
| WB 1,
4.87 | 0.034 | 0.7 | 0.004 | 0.1 | 0.036 | 0.7 | 0.040 | 0.8 | 0.007 | 0.1 | 0.064 | 1.3 | |
| WB 2,
6.57 | 0.029 | 0.4 | 0.012 | 0.2 | 0.028 | 0.4 | 0.075 | 1.1 | 0.024 | 0.4 | 0.090 | 1.4 | |
| WB 3,
7.34 | 0.037 | 0.5 | 0.009 | 0.1 | 0.032 | 0.4 | 0.087 | 1.2 | 0.027 | 0.4 | 0.104 | 1.4 | |
| WB 4,
8.20 | 0.041 | 0.5 | 0.011 | 0.1 | 0.042 | 0.5 | 0.096 | 1.2 | 0.031 | 0.4 | 0.117 | 1.4 | |
| WB 5,
12.54 | 0.069 | 0.5 | 0.020 | 0.2 | 0.062 | 0.5 | 0.155 | 1.2 | 0.043 | 0.3 | 0.187 | 1.5 | |
| WB_006,
14.64 | 0.086 | 0.6 | 0.028 | 0.2 | 0.096 | 0.7 | 0.181 | 1.2 | 0.043 | 0.3 | 0.228 | 1.6 | |
| WB_007,
12.27 | 0.070 | 0.6 | 0.023 | 0.2 | 0.055 | 0.5 | 0.159 | 1.3 | 0.055 | 0.4 | 0.192 | 1.6 | |
| WB_008,
13.08 | 0.073 | 0.6 | 0.000 | 0.0 | 0.061 | 0.5 | 0.161 | 1.2 | 0.049 | 0.4 | 0.194 | 1.5 | |
| PCA1N,
5.51 | 0.032 | 0.6 | 0.002 | 0.0 | 0.032 | 0.6 | 0.047 | 0.8 | 0.007 | 0.1 | 0.065 | 1.2 | |
| PCA1P,
11.14 | 0.063 | 0.6 | 0.019 | 0.2 | 0.054 | 0.5 | 0.141 | 1.3 | 0.047 | 0.4 | 0.171 | 1.5 | |

Table 9: Precision Results – Whole Blood Application, 3 Lots and 3 Analytical Units

Analytical Sensitivity 4.2.

Limit of Blank (LoB) 4.2.1.

For determination of LoB, one analyte free sample was measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3, in 10-fold determinations. Six runs were distributed over ≥ three days and measured using one cobas c 503 analytical unit. In total, 60 measurements were obtained per lot. Data analysis was based on determination of the 95th percentile of the 60 measured values. In our design (n=60) the 95th percentile is the average of the 57th and 58th value.

LoB was determined according to CLSI guideline EP17-A2.

Table 10: LoB Results

	Hb LoB		HbA1c LoB
	mmol/L	g/dL	mmol/L	g/dL
	0.0530 mmol/L	0.085 g/dL	0.0220 mmol/L	0.035 g/dL

This corresponds to a Limit of Blank of 15 mmol/mol (IFCC) and 3.5 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).

14

Limit of Detection (LoD) 4.2.2.

For determination of LoD, five unique human samples with low-analyte concentrations were measured with three lots of Tina-quant Hemoglobin A1cDx Gen.3 in two-fold determinations. The measurements were performed in six runs, over > three days, on one cobas c 503 analytical unit. In total. 60 measurements were obtained per lot.

LoD is defined as the concentration, at which there is a 95% probability that a sample contains analyte.

LoD was determined according to CLSI guideline EP17-A2.

Table 11: LoD Results

Hb LoD		HbA1c LoD
mmol/L	g/dL	mmol/L	g/dL
0.119 mmol/L	0.192 g/dL	0.0437 mmol/L	0.07 g/dL

This corresponds to a Limit of Detection of 22 mmol/mol (IFCC) and 4.2 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).

Linearity/Assay Reportable Range 4.3.

Separate dilution series, consisting of at least eleven levels, were prepared for each glycated hemoglobin (HbA1c) and total hemoglobin (Hb) using human hemolysate sample pools. The sample pools include HbA1c and Hb concentrations above the upper end of the corresponding measuring range. Hemolyzing reagent was used for the diluent. Samples were measured in triplicate and data analysis was performed separately for each sample.

The study was performed according to CLSI guideline EP6-A.

Table 12: Linearity Results according to CLSI EP6-A

Application	Analyte	Low End of Linear Range		High End of Linear Range
		g/dL	mmol/L	g/dL	mmol/L
Hemolysate	Hb	3.04	1.89	40.4	25.1
	HbA1c	0.293	0.182	2.87	1.78

15

Table 13: Empirical First Order Regression Results

Application	Analyte	Slope	Intercept	Pearson's r
Hemolysate	Hb	1.019	-0.1552	0.9999
	HbA1c	0.991	-0.0026	0.9990

Linearity was determined throughout the claimed measuring range of:

4 – 40 g/dL (2.48 – 24.8 mmol/L) Hb:

0.3 – 2.6 g/dL (0.186 – 1.61 mmol/L) HbA1c:

This corresponds to a measuring range of 23-196 mmol/mol HbA1c (IFCC) and 4.2-20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 13.2 g/dL (8.2 mmol/L).

4.4. Endogenous Interferences

A study evaluated several endogenous substances for potential interference with measurement of % HbA1c. The following nine endogenous substances were evaluated.

Bilirubin .
Ditaurobilirubin .
Lipemia .
Rheumatoid Factors .
Total Protein .
Albumin .
Immunoglubulin (IgG) .
Glucose .
Triglycerides .

Pooled whole blood samples, with two hemoglobin A1c levels, one near the medical decision level and one above it, were spiked with the maximum level of the above nine interferents, in separate preparations, resulting in eighteen spiked samples were then hemolyzed

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with Tina-quant HbA1c Hemolyzing Reagent. Another pool, without interferent, was equally hemolyzed. A > ten-level dilution series was created for each of the eighteen spiked samples, using the interferent-free pools as the diluent.

The eighteen dilution series were tested in ten-fold, using one reagent lot, one cobas c 503 analytical unit, in a single run and within one calibration cycle. Additionally, PreciControl HbA1c norm and PreciControl HbA1c path were used as the controls. The mean of the ten replicates was compared to the result from the reference sample (aliquot with no interferent). The comparison was evaluated as a percent deviation. For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.

Potential Interferent	Claimed Maximum Concentration without Interference
Bilirubin	60 mg/dL
Ditaurobilirubin	60 mg/dL
Lipemia	400 mg/dL
Rheumatoid Factors	750 IU/mL
Total Protein	21 g/dL
Albumin	60 g/L
Immunoglobulin (IgG)	60 g/L
Glucose	1000 mg/dL
Triglycerides	1584 mg/dL

Table 14: Endogenous Interference

4.5. Cross-Reactivity

This study was conducted to evaluate the Tina-quant Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit for potential cross-reactivity with the following hemoglobin fractions and glycated albumin.

HbA0 .
Carbamylated Hb .
HbA1(a+b) .
Acetylated Hb .
Labile HbA1c Glycated Albumin . .

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A series of experiments were performed using one reagent lot, on one cobas c 503 analytical unit, in a single run, within one calibration cycle. PreciControl HbA1c norm and PreciControl HbA1c path were used as the controls. Ten replicates of each sample were analyzed for each dilution level. The median % HbA1c of each dilution level was compared to the median % HbA1c from dilution level zero (without cross-reactant). For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.

| Cross-Reactant | | Max Whole Blood Cross-
Reactant Concentration | Max Whole Blood Cross-
Reactant Concentration with
no Interference |
|------------------|---------------|--------------------------------------------------|--------------------------------------------------------------------------|
| HbA0 | HbA1c Level 1 | 120 g/dL | 120 g/dL |
| | HbA1c Level 2 | 120 g/dL | 120 g/dL |
| HbA1(a+b) | HbA1c Level 1 | 1.6 g/dL | 0.96 g/dL |
| | HbA1c Level 2 | 1.6 g/dL | 1.6 g/dL |
| Carbamylated Hb | HbA1c Level 1 | 2.0 g/dL | 2.0 g/dL |
| | HbA1c Level 2 | 2.0 g/dL | 2.0 g/dL |
| Acetylated Hb | HbA1c Level 1 | 2.0 g/dL | 2.0 g/dL |
| | HbA1c Level 2 | 2.0 g/dL | 2.0 g/dL |
| Glycated Albumin | HbA1c Level 1 | 10 g/dL | 10 g/dL |
| | HbA1c Level 2 | 10 g/dL | 10 g/dL |
| Labile HbA1c | HbA1c Level 1 | 1000 mg/dL | 1000 mg/dL |
| | HbA1c Level 2 | 1000 mg/dL | 1000 mg/dL |

Table 15: Cross-Reactivity

Hemoglobin Variants 4.6.

Hemoglobin variant testing was conducted to determine if significant interference with any of the major hemoglobin variants occurred when using the Tina-quant Hemoglobin A1cDx Gen.3 assay on cobas c 503 analytical unit. Hemoglobin variants are structurally altered hemoglobin molecules with at least one amino acid exchange, compared to the normal beta chain of hemoglobin. These changes are caused by mutations in the coding region of the globin genes which encode the protein part of hemoglobin. The most common hemoglobin variants are HbS, HbC, HbD and HbE. Additionally, in some conditions fetal hemoglobin, HbF, is elevated. Also, the erythrocytes of some patients (e.g. beta thalassemia minor) contain elevated levels of HbA2.

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Therefore, it is crucial to ensure accurate HbA1c results from patients who are carriers of these variants.

Variant Type	Number of Samples	% Variant	HbA1c %
HbS	30	35-41% S	4.35 - 12.7
HbC	30	28-37% C	4.90 - 14.1
HbE	30	24-27% E	5.17 - 10.0
HbD	29	36-42% D	5.17 - 9.70
HbA2	15	4.3-6.5% A2	5.10 - 9.80
Elevated HbF	19	3.2-39% F	6.10 - 9.30

Table 16: Hemoglobin Variant Samples

Each sample was tested once, in at least one run, on one cobas c 503 analytical unit. Results obtained with the Tina-quant Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit were compared to those obtained with the corresponding reference method. For purposes of this experiment, the data was collected using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.

Table 17: Hemoglobin Variant Testing

Percent Relative Bias from Reference Method at Low and High Concentrations of HbA1c Samples
	Around 6.5% HbA1c		Around 9% HbA1c
HbVariant	Relative % Difference	Range	Relative % Difference	Range
HbS	-2.5	-7.2 - 3.2	-4.0	-9.3 - (-2.0)
HbC	-3.9	-7.7 - 2.8	-6.0	-4.6 - (-3.6)
HbE	-0.1	-5.5 - 5.7	-1.2	-5.2 - 0.6
HbD	-1.8	-4.5 - 3.0	-2.6	-3.3 - 0.2
HbA2	-1.0	-4.1 - 2.7	0.4	-2.2 - 1.1
HbF	Specimens containing high amounts of HbF (>7%) may yield lower than expected HbA1c values.

Exogenous Interferences – Drugs 4.7.

The purpose of this study was to evaluate drugs for potential interference with the Tina-quant Hemoglobin A1cDx Gen.3 assay measured on the cobas c 503 analytical unit.

The eighteen commonly used drugs listed below were added to samples and examined for potential effect on % HbA1c determination. Drug interference testing was performed with

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hemolysate samples at two different HbA1c levels, approximately 6% and 9% HbA1c. Each drug was added in two defined concentrations with concentration one being several times (typically five times) the maximum daily dosage and concentration 2 being the maximum daily dosage level. Concentration one was performed for screening purposes only and concentration two was the relevant drug concentration for determining interferences with the assay. Samples were measured in ten-fold using the cobas c 503 analytical unit. The median value was compared to the reference value (HbA1c sample with no drug added) and the deviation from the reference was calculated. Drug interference studies were conducted using the Hemolysate Application and were representative of both Hemolysate and Whole Blood Applications.

. N-Acetylcysteine
Acetylsalicylic acid .
Ampicillin-Na .
Ascorbic acid .
Cefoxitin .
Heparin .
Levodopa .
Methyldopa + 1.5 .
Metronidazole .
Doxycyclin .
Rifampicin .
Gammagard .
Cyclosporine .
Phenylbutazone .
Acetaminophen .
Ibuprofen .
Theophylline .
Tolbutamide .

Sample Matrix Comparison 4.8.

The purpose of this study was to evaluate Hemoglobin A1c determination, with the Tina-quant Hemoglobin A1cDx Gen.3 assay, in the presence of anticoagulants.

At least 40 samples of each anticoagulant and at least 40 half-filled tubes of each anticoagulant were evaluated. The filled and corresponding half-filled (double concentrated) sample tubes were from one donor. Matrix comparison studies were conducted using the Hemolysate Application and was representative of both Hemolysate and Whole Blood Applications.

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| Sample Type | Anticoagulant | Tube Fill | Mean
Difference | Upper 95% | Lower 95% |
|-------------|--------------------------|-----------|--------------------|-----------|-----------|
| Hemolysate | K2-EDTA | ½ Full | 0.004 | 0.124 | -0.116 |
| | K3-EDTA | Full | -0.005 | 0.104 | -0.114 |
| | | ½ Full | 0.004 | 0.140 | -0.131 |
| | Na Heparin | Full | 0.000 | 0.120 | -0.121 |
| | | ½ Full | -0.026 | 0.129 | -0.181 |
| | Li Heparin | Full | -0.003 | 0.081 | -0.088 |
| | | ½ Full | -0.015 | 0.138 | -0.167 |
| | NaF/Potassium
oxalate | Full | 0.009 | 0.123 | -0.105 |
| | | ½ Full | 0.019 | 0.135 | -0.097 |
| | EDTA/Fluoride | Full | 0.003 | 0.100 | -0.094 |
| | | ½ Full | 0.017 | 0.172 | -0.139 |

Table 18: Matrix Comparison Results

4.9. Method Comparison

A method comparison study was performed to compare the sample results from the candidate method, Hemoglobin A1cDx Gen.3 assay on the cobas c 503 analytical unit, to results from Tosoh Automated Glycohemoglobin Analyzer HLC-723G8 (Tosoh), the NGSP method. This study was conducted with both the Tina-quant Hemoglobin A1cDx Gen.3 Whole Blood Application and Hemolysate Application.

One hundred and seventy-one whole blood samples and one hundred seventy-three hemolysate samples from the secondary NGSP reference laboratory were used in the evaluation. These samples were measured by the secondary NGSP reference laboratory using the Tosoh HPLC system (X-axis) and the Roche Tina-quant Hemoglobin A1cDx Gen.3 test system (Y-axis). The samples were tested over a 3-day period, with one lot of reagent, on one cobas c 503 analytical unit.

	# Samples Tested		% Samples Tested
% HbA1c	Whole Blood	Hemolysate	Whole Blood	Hemolysate
≤ 5%	6	6	3.5%	3.5%
5-6 %	23	24	13.5%	13.9%

Table 19: Method Comparison Sample Distribution

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>6 - 6.5%	31	32	18.1%	18.5%
>6.5 - 7%	37	37	21.6%	21.4%
>7 – 8%	25	25	14.6%	14.5%
>8 – 9%	13	13	7.6%	7.5%
>9%	36	36	21.1%	20.8%
Total	171	173	100%	100%

The method comparison demonstrated good agreement between Roche Tina-quant Hemoglobin A1cDx Gen.3 assay and the NGSP Tosoh reference method. The tables below summarize the bias between the Tina-quant Hemoglobin A1cDx Gen.3 assay and the NGSP Tosoh reference method.

Table 20: Difference Plot Analysis Data Summary

	Whole Blood Application	Hemolysate Application
Mean bias vs. NGSP TOSOH	-0.046%	0.046%
Mean bias at lower 95% CI	-0.410%	-0.338%
Mean bias at upper 95% CI	0.318%	0.431%

Table 21: Bias at Concentration Data Summary

| % HbA1c | % Relative Bias
Whole Blood Application | % Relative Bias
Hemolysate Application |
|---------|--------------------------------------------|-------------------------------------------|
| 5% | -2.4% | 0.6% |
| 6.5% | -1.2% | 0.8% |
| 8% | -0.4% | 1.0% |
| 12% | 0.7% | 1.2% |

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4.10. Total Error

Using the results of bias estimation (%Bias) generated during the method comparison study and precision estimates (%CV) from the precision study, the Total Error (TE) was calculated using the following equation:

%TE = |%Bias| + 1.96 * %CV * (1 + %Bias/100).

Table 22: Total Error – Hemolysate Application

%HbA1c	% BIAS	Precision (%CV)	Total Error (%)
4.96	0.63	1.2	3.0
6.59	0.87	1.2	3.3
8.29	1.02	1.3	3.5
12.1	1.20	1.6	4.4

Table 23: Total Error – Whole Blood Application

%HbA1c	% BIAS	Precision (%CV)	Total Error (%)
4.87	-2.56	1.3	5.2
6.57	-1.12	1.4	3.8
8.20	-0.30	1.4	3.1
12.3	0.79	1.6	3.9

ADDITIONAL INFORMATION 5.

Other Devices Marketed with This Assay 5.1.

The following devices are required, but not provided:

C.f.a.s. (Calibrator for automated systems) HbA1c, K052101 .
PreciControl HbA1c norm, K103099 .
PreciControl HbA1c path, K103099 .

CONCLUSIONS 6.

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