Predicate Devices

COBAS INTEGRA 800 Tina-quant HbA1cDx Gen.2 assay, K121291

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on standard automated clinical chemistry analysis using photometric methods and software control for instrument functions and data management. There is no mention of AI/ML algorithms for data analysis, interpretation, or decision support. The performance studies are based on traditional analytical validation methods.

Therapeutic

No
This device is an in-vitro diagnostic device used for quantitative determination of analytes to aid in the diagnosis and monitoring of diabetes, not a device that directly treats or prevents a disease.

Diagnostic

Yes

The Tina-quant Hemoglobin A1cDx Gen.3 assay, which is used with the cobas c 513 clinical chemistry analyzer, is explicitly stated as being "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 indicates its role in the diagnostic process.

SaMD (Software as a Medical Device)

No

The device is a clinical chemistry analyzer system which includes significant hardware components (analyzer, control unit, sampler unit, analytical unit) in addition to the software that controls it.

IVD (In Vitro Diagnostic)

Based on the provided information, the device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use / Indications for Use: Explicitly states "intended for the in-vitro quantitative determination of analytes in body fluids" and "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 whole blood".
Device Description: Describes a system designed for "in vitro quantitative determination of analytes in human body fluids".
Predicate Device: A predicate device (K121291; COBAS INTEGRA 800 Tina-quant HbA1cDx Gen.2 assay) is listed, which is also an IVD.

All these points clearly indicate that the device is intended for use outside of the body to examine specimens for diagnostic purposes, which is the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The cobas c 513 clinical chemistry analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.

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 whole blood on the Roche/Hitachi cobas c 513 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, JJE

Device Description

Roche Diagnostics plans to introduce the cobas c 513 clinical chemistry analyzer, a new member of the Roche / Hitachi family of analyzers. It has been developed jointly by Roche Diagnostics and Hitachi High Technologies (HHT). It is a standalone, high volume Roche/Hitachi cobas c analyzer, that is to be used with the Tina-quant HbA1cDx Gen.3 Assay which is intended for use with human whole blood and hemolysate samples for monitoring of long term blood glucose control in individuals with diabetes mellitus and as "an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes".

Analyzer
The cobas c 513 analyzer is a fully automated, software controlled analyzer system for in vitro quantitative determination of analytes in human body fluids. The cobas c 513 analyzer system includes a control unit and an analyzer with a closed tube sampling functionality.

The cobas c 513 control unit includes a computer (PC) located in the sampler unit, a touchscreen monitor, soft-keyboard, a mouse (optional use), and a printer. The control unit is used to perform tasks on the analyzer and the PC runs the software that controls the analyzer.

The software manages all instrument functions, all system functions, and all information related to orders and results. The software offers a graphical user interface (GUI) to control all functions by the operator. The control unit contains System Control software including interfaces to a Medical Device Data System (cobas Link) and to the customer Laboratory Information System (LIS).

The analyzer is composed of the sampler unit and the analytical unit. The sampler unit is composed of the rack loading/unloading areas, a barcode reader, a rack rotor, a STAT port, and conveyor lines. It is used to load and unload racks, power on the system, access the PC, and manage the conveyance of samples to the analytical unit.

The analytical unit is comprised of the reagent area, the sample area, and the reaction disk. Samples are conveyed to the analytical unit from the sample unit for photometric analysis before being returned to the sample unit.

Assay
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.

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. Hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer. Hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results.

Test principle
This method uses tetradecyltrimethyl-ammonium bromide (TTAB) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). 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. Sample pretreatment to remove labile HbA1c is not necessary. All hemoglobin variants which are glycated at the ßchain 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.

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 R2 (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.

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.

The final result is expressed as mmol/mol HbA1c or % 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

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

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

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Description of the test set, sample size, data source, and annotation protocol

A method comparison study was performed to compare the sample results from the candidate method, Tina-quant HbA1c Gen.3 on the cobas c 513, to results from the TOSOH 8 HPLC. This study will be conducted with both the Tina-quant HbA1c Gen.3 Whole Blood Application and Hemolysate Application.

One hundred and fifty-five (155) samples from the secondary NGSP reference laboratory were used in the evaluation. These samples were measured by the secondary NGSP reference laboratory using a Tosoh HPLC system (X axis) and by the Roche Tina-quant HbA1c Gen.3 test system (Y axis). Samples were tested over a 3 day period with one lot of reagent on one cobas c 513 analyzer.

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

Non-Clinical Performance Evaluation:
Performance characteristics were evaluated with the Tina-quant HbA1c Gen.3 reagent on the cobas c 513 analyzer. This assay offers two sample type-specific applications, one for manually hemolysed samples and one for whole blood samples; thus they are named the Hemolysate and Whole Blood Applications.

The Tina-quant HbA1c Gen.3 assay first measures total hemoglobin (Hb) and glycated hemoglobin (HbA1c) in terms of mmol/L. Then the analyzer calculates the HbA1c/Hb ratio according to the corresponding protocol, 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 include limit of detection and linearity. They report results in terms of Hb and HbA1c individually. A patient sample value is reported in terms of the ratio of glycated to total hemoglobin. Method comparison, control recovery, and precision are evaluated in terms of the ratio. The protocol according to DCCT/NGSP was chosen for these studies.

4.1. Limit of Blank and Limit of Detection (LoB and LoD)
Limit of Blank: For determination of LoB one analyte free sample will be measured with three lots in 10-fold determination. Six runs distributed over > 3 days using one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot. Data analysis will be based on determination of the 95th percentile of the 60 measured values.
Results: Hb: 0.50 g/dL (0.31 mmol/L), HbA1c: 0.19 g/dL (0.12 mmol/L).

Limit of Detection: For determination of LoD, five samples with low-analyte concentration will be measured with three lots in two-fold determination. Six runs distributed over > three days on one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot. LoD is defined as the concentration, at which there is a 95% probability that a sample contains analyte.
Results: Hb: 1.00 g/dL (0.62 mmol/L), HbA1c: 0.29 g/dL (0.18 mmol/L).

4.2. Precision
Precision experiments were performed in Accordance with CLSI Guideline EP5-A2 to evaluate repeatability (within-run precision) and intermediate precision of within-laboratory precision (total precision).
Two aliquots per sample were measured once each in two runs per day for 21 days on three cobas c 513 analyzers using 3 reagent lots per system. Eight total samples were evaluated in each run: two controls, PreciControl HbA1c norm and path, and 7 human samples with HbAc concentrations near 5%, 6.5%, 7.0%, 8.0%, 10.5%, 12%, and 14%.
The samples were randomized in each run separately. The data set was completed for the 21 days. 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.
Key Results: Precision for both Hemolysate and Whole Blood applications showed low CVs, generally below 1.5% for total precision across various HbA1c concentrations.

4.3. Method Comparison
A method comparison study was performed to compare the sample results from the candidate method, Tina-quant HbA1c Gen.3 on the cobas c 513, to results from the TOSOH 8 HPLC. This study was conducted with both the Tina-quant HbA1c Gen.3 Whole Blood Application and Hemolysate Application.
One hundred and fifty-five (155) samples from the secondary NGSP reference laboratory were used. Samples were tested over a 3 day period with one lot of reagent on one cobas c 513 analyzer.
Key Results: All acceptance criteria for method comparison were met. The difference plots show good agreement between the Roche method and the NGSP TOSOH HPLC reference method.
Mean bias (vs. NGSP TOSOH): Whole Blood Application: -0.016; Hemolysate Application: -0.138.
Percent Relative Bias at %>5%: Whole Blood Application: 0.1%; Hemolysate Application: -0.8%.

4.4. Total Error Near the Cut-off
Total Error (TE) was calculated using bias estimation (%Bias) from the method comparison study and precision estimates from the precision study: %TE = |%Bias| + 1.96 * %CV * (1+%Bias).
Key Results:
Whole Blood Application: TE ranged from 2.1% (at 6.5% HbA1c) to 3.9% (at 5.0% HbA1c).
Hemolysate Application: TE ranged from 3.7% (at 5.0% and 6.5% HbA1c) to 5.2% (at 12.0% HbA1c).

4.5. Linearity
Two separate dilution series consisting of at least 11 levels were prepared using human hemolysate sample pools with concentrations above the upper end of the measuring range. Hemolysing reagent was used for the diluent. Samples were measured in triplicate and data analysis was done separately for each sample. Linear regression analysis was done according to EP6-A.
Key Results:
For Hemolysate:
Hb: Slope = 0.983, Intercept = 0.051, Pearson's r = 0.9999. Claimed Range: 4.0 - 40 g/dL (2.48-24.8 mmol/L).
HbA1c: Slope = 0.972, Intercept = 0.002, Pearson's r = 0.9990. Claimed Range: 0.3 - 1.93 g/dL (0.186-1.20 mmol/L).
This corresponds to a measuring range of 23-146 mmol/mol HbA1c (IFCC) and 4.2-15.5 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L.

4.6. Matrix Comparison
A matrix comparison study was performed to evaluate different anticoagulants on analyte recovery. At least 40 samples of each sample type and at least 40 half-filled tubes of each sample type were evaluated. The Hemolysate application was used.
Key Results: All acceptance criteria for matrix comparison were met. Mean differences were small, e.g., K2-EDTA (½ full) -0.0559, K3-EDTA (Full) 0.000732, Li Heparin (Full) 0.0156.

4.7. Endogenous Interference
A study evaluated six endogenous substances (Lipemia, Bilirubin, Ditaurobilirubin, Glucose, Rheumatoid Factor, Total Protein) for potential interference with % HbA1c. Samples were spiked and results compared to interferent-free pools.
Key Results: Interference was considered significant if causing >7% deviation. Maximum concentrations without interference were reported: Lipemia 600 mg/dL, Bilirubin 60 mg/dL, Ditaurobilirubin 60 mg/dL, Glucose 1000 mg/dL, Rheumatoid Factor 750 IU/mL.

4.8. Drug Interferences
A study evaluated 16 commonly used drugs for potential interference with %HbA1c measurement using the hemolysate application. Drugs were added at two concentrations to samples with two different HbA1c levels (~6% and ~8%).
Key Results: All acceptance criteria for drug interferences were met. Significant interference was defined as ≥ ±7% deviation from the reference value.

4.9. Cross reactivity
Studies were performed to determine if the Tina-quant Hemoglobin A1cDx Gen.3 assay demonstrates cross-reactivity with HbA0, HbA1a+b, Acetylated Hb, Carbamylated Hb, Glycated Albumin, and Labile HbA1c.
Key Results: Cross-reactivity was evaluated by % recovery. Maximum concentrations with no interference were reported for each cross-reactant, indicating minimal or no cross-reactivity at tested concentrations.

4.10. Hemoglobin Variants
Hemoglobin variant testing was conducted to determine if there was any significant interference from HbS, HbC, HbD, HbE, Elevated F, and HbA2. Samples were tested once on one c 513 analyzer, comparing results to reference methods.
Key Results: No significant interference was found with most major hemoglobin variants (HbAS, HbAD, HbAE). Significant interference was defined as ≥7% change in HbA1c value. Specimens with high amounts of HbF (>7%) may yield lower than expected HbA1c values. Relative % differences from reference method were generally low (e.g., HbA2: -2.0% at ~6.0% HbA1c)

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

Not Found

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 HbA1cDx 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/1 description: The image is a black and white logo for the Department of Health & Human Services - USA. The logo features a stylized image of three human profiles facing to the right, stacked on top of each other. The profiles are connected and form a single, abstract shape. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the image.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

December 19, 2016

ROCHE DIAGNOSTICS OPERATIONS DAVID TRIBBETT REGULATORY PROGRAM MANAGER 9115 HAGUE ROAD INDIANAPOLIS, IN 46250

Re: K160571 Trade/Device Name: cobas c 513 Analyzer, cobas c 513 Tina-quant HbA1cDx Gen.3 Assay Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin Alc test system Regulatory Class: II Product Code: PDJ, JJE Dated: December 08, 2016 Received: December 09, 2016

Dear David Tribbett:

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. 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Courtney

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology 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) K160571

Device Name cobas c 513 Analyzer cobas c 513 Tina-quant HbA1cDx Gen.3 Assay

Indications for Use (Describe)

The cobas c 513 clinical chemistry analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.

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 Alc (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate or whole blood on the Roche/Hitachi cobas c 513 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)

Registration Use (Part 81 CFR 801 Subpart D)
Over-The-Counter Use

|X Prescription Use (Part 21 CFR 801 Subpart D)

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

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cobas c 513 Tina-quant Hemoglobin A1cDx Gen.3 assay 510(k) Summary

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

Submitter Name	Roche Diagnostics Operations (RDO)
Address	9115 Hague Road
Indianapolis, IN, 46250, USA
Contact	David Tribbett
Phone: (317)-521-2964
FAX: (317)-521-2324
Email: david.tribbett@roche.com
Date Prepared	January 31, 2016
Proprietary Names	cobas c 513 clinical chemistry analyzer
Tina-quant Hemoglobin A1cDx Gen.3
Common Names	Analyzer, Chemistry (Photometric, Discrete), for clinical use
Glycosylated Hemoglobin Assay
Classification Name	Discrete photometric chemistry analyzer for clinical use
Hemoglobin A1c test system
Product Codes	JJE, 21 CFR 862.2160
PDJ, 21 CFR 862.1373
Predicate Device	COBAS INTEGRA 800 Tina-quant HbA1cDx Gen.2 assay, K121291
Establishment Registration	1823260, Roche Diagnostics Corporation

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1. DEVICE DESCRIPTION

Roche Diagnostics plans to introduce the cobas c 513 clinical chemistry analyzer, a new member of the Roche / Hitachi family of analyzers. It has been developed jointly by Roche Diagnostics and Hitachi High Technologies (HHT). It is a standalone, high volume Roche/Hitachi cobas c analyzer, that is to be used with the Tina-quant HbA1cDx Gen.3 Assay which is intended for use with human whole blood and hemolysate samples for monitoring of long term blood glucose control in individuals with diabetes mellitus and as "an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes".

Analyzer

The cobas c 513 analyzer is a fully automated, software controlled analyzer system for in vitro quantitative determination of analytes in human body fluids. The cobas c 513 analyzer system includes a control unit and an analyzer with a closed tube sampling functionality.

The cobas c 513 control unit includes a computer (PC) located in the sampler unit, a touchscreen monitor, soft-keyboard, a mouse (optional use), and a printer. The control unit is used to perform tasks on the analyzer and the PC runs the software that controls the analyzer.

The software manages all instrument functions, all system functions, and all information related to orders and results. The software offers a graphical user interface (GUI) to control all functions by the operator. The control unit contains System Control software including interfaces to a Medical Device Data System (cobas Link) and to the customer Laboratory Information System (LIS).

The analyzer is composed of the sampler unit and the analytical unit. The sampler unit is composed of the rack loading/unloading areas, a barcode reader, a rack rotor, a STAT port, and conveyor lines. It is used to load and unload racks, power on the system, access the PC, and manage the conveyance of samples to the analytical unit.

The analytical unit is comprised of the reagent area, the sample area, and the reaction disk. Samples are conveyed to the analytical unit from the sample unit for photometric analysis before being returned to the sample unit.

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Assay

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.

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. Hemolysis occurs onboard the analyzer. For the Hemolysate Application, hemolyzed samples are placed on the analyzer. Hemolysis occurs manually before placing the samples onboard the analyzer. The two applications yield the same results.

Test principle

This method uses tetradecyltrimethyl-ammonium bromide (TTAB) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). 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. Sample pretreatment to remove labile HbA1c is not necessary. All hemoglobin variants which are glycated at the ßchain 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.

Hemoglobin A1c

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

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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 R2 (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.

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.

The final result is expressed as mmol/mol HbA1c or % 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

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.

2. INDICATIONS FOR USE

The cobas c 513 analyzer is a fully automated, standalone clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.

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

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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 Roche/Hitachi cobas c 513 clinical chemistry analyzer. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus.

3. TECHNOLOGICAL CHARACTERISTICS

As the cobas c 513 analyzer is a new member of the Roche/Hitachi clinical chemistry instruments family, a comparison to the cobas c 501 analyzer is shown in the table below to compare the analyzer to other members of the Roche/Hitachi Clinical Chemistry instrument family.

Topic	cobas c 501	cobas c 513
	Basic features
Intended Use	Fully automated clinical chemistry analyzer intended for the in vitro quantitative / qualitative determination of analytes in body fluids	The cobas c 513 is a fully automated, discrete clinical chemistry analyzer intended for the in-vitro quantitative determination of analytes in body fluids.
Measurement principle	Absorbance Photometry (enzymes, substrates, proteins, DAT, TDM)	Absorbance Photometry (enzymes, substrates, proteins, DAT, TDM)
	ISE Potentiometric (electrolytes)
Endpoint, Kinetic,	N/A
Reaction modes	Potentiometric	Same
		N/A
Throughput Photometry	Max 600 tests per hour without ISE	400 tests per hour
Throughput ISE Potentiometry	Max 600 tests per hour	N/A
Analyzer	Standalone module or multiple modules linked	Standalone analyzer
	Sample Handling
Typical sample volumes	1.0 – 35 μL	S1: 1.5-25 μl
S2: 1.5-6 μl
Sample types	Serum, plasma, urine, CSF and other depending on the chemistry test	Hemolysate, whole blood, Serum/Plasma, Urine
Sample handling system	Input of samples via core input buffer using universal sample racks	Closed Tube Sampling (CTS) on specified 5 position Racks ( CTS Rack) and universal sample racks
Sample capacity On board	150	Same
Sample identification	Barcode	Same
Topic	cobas c 501	cobas c 513
	Reagent Handling
Reagent volume	5 – 180 μL	15-150 μl
Reagent container
(electrolytes)	Plastic bottles open	N/A
Reagent container
(non-electrolytes)	Plastic bottles closed via pierceable screwcaps	Plastic bottles closed via screwcaps
The cap of the reagent bottle is
decapped (opened) by operator
Reagent access	Reagent cassette caps pierced onboard by the
instrument	Cassette caps to be opened before
placing on instrument
Onboard storage
Temperature	Refrigerated
5 -12 °C	Refrigerated
5-15°C
Reagent bottle / Cassette
identification	Barcode	RFID
On board reagent storage
capacity	60 rotor channels on 1 rotor to run 60 reagent kits
in parallel	Same
System cycle time	6 sec	7.2 sec
Reagent mixing	Ultrasonic	Same
Auto rerun	Available	Same
Application information
transfer to instrument	Via remote transfer or CD	Via remote transfer
	Pipetting System
Sample and reagent
Syringes	XY robotic	Same
Reagent probes	2 polished steel probes	Same
Sample probes	1 polished steel probe	2 polished steel probes
Probe cleaning	Automatic for all probes	Same
Liquid level detection (LLD)
sample	Electrostatic	Electrostatic for open tubes (S1), no
LLD for CTS (S2)
Liquid level detection
reagent	Initial Cassette Volume Check (ICVC)	Electrostatic (Capacitance)
Clot detection	Provided	Same
	Test Reaction Chamber
Temperature control	Circulating water bath at 37°C	Same
Cuvettes	Multiple use	Same

	Detection Information
Spectrophotometer	Gradient photometer with discrete photodiodes in
fixed array	Same
Light source	Tungsten/halogen	Same
Light path	5.6 mm	5 mm
Measuring unit	1	Same
Wavelengths	340,376,415,450,480,505,546, 570, 600,
660,700,800 nm	Same
	Calibration and Quality Control
Calibrators	Multiple use	Same
Calibration modes	Linear, nonlinear	Same
Calibration Stability	Typically each lot or 12 weeks for same reagent
on board	For HbA1c: 4 weeks during shelf live
Online QC	Yes	No automatic QC, but QC timeout
function
Control storage on
instrument	In remote buffer at ambient temperature	none
Calibrator/control value
transfer	Via remote transfer or CD	Via remote transfer
Topic	cobas c 501	cobas c 513
Internal quality
management system	Available to monitor and validate test results	Same
Software and Interfaces
Host Interface	RS232C bi-directional	LAN/Ethernet
Printer	Laser	Any printer supported by the OS
Display	Keyboard + touch-screen
Optional use of mouse	Keyboard + touch-screen: soft-
keyboard; Optional use of mouse
Software	cobas 6000 modular system software	c 513 instrument software
Configuration	One or several analytical units with one PC and
one Core	One stand-alone analytical unit with
one PC
Units controlled	cobas c501 and cobas e601	513 analytical unit, one unit
Functions performed	Data input, sample processing, result calculation,
result reporting, quality control	Same
PC (Controller Unit)
functions	Data input (keyboard, disc),
Data output (screen, printer)	Data input (keyboard, disc)
no input from USB,DVD-RAM
Data output (screen, printer + USB DVD-RAM)
Core Unit functions	Realtime database, data input and output ( via
HOST Communication ) control of sample conveyer	Same
Analytical Unit(s) function	Control of analytic processes (pipetting, incubation,
detection) Primary Signal processing	Same
Data storage	Real time database in Core unit
(storage of System and Application parameters,
Calibration Data ,QC Data, Sample results, Alarm
history)	Same
Software-controlled test
countdown	Available	Same
Result calculation	Automated measuring of signal for kinetic and
endpoint methods according to cycle time and
automated calculation of concentrations via
calibration curve	Same
User management	Yes	Same
Flagging of errors	Available	Same

Substantial Equivalence – Analyzer Similarities

8

9

Substantial Equivalence – Assay Similarities

While the c 513 analyzer is a new member of the Roche /Hitachi family of analyzers, none of the analyzers in this family have received 510(k) clearance for an HbA1c "aid in diagnosis" assay for both human whole blood and hemolysate sample types. Sample type is a key element of the intended use, which necessitates the COBAS Integra 800 Tina-quant Hemoglobin A1cDx Gen.2 to be the predicate device since it has been cleared for aid in diagnosis for both of these sample types.

10

| Feature | Predicate Device: COBAS INTEGRA 800
Tina-quant HbA1cDx Gen.2 assay
K121291 | Candidate Device: cobas c 513 Tina-
quant HbA1cDx Gen.3 assay |
|--------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 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. | This test 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 whole blood on the
Roche/Hitachi cobas c 513 clinical
chemistry analyzer. HbA1c
determinations are useful for monitoring of
long-term blood glucose control in
individuals with diabetes mellitus. |
| Sample Types | Anticoagulated venous or capillary blood

Acceptable anticoagulants for both the
hemolysate and whole blood applications
include:
Li-Heparin
K2-EDTA
K3-EDTA
Na-Heparin
NaF/K-Oxalate
NaF/Na2-EDTA | Anticoagulated venous blood

On-bard in use:
• 2-8°C for 28 days | Same |
| Reporting Units | % HbA1c NGSP/DCCT and mmol/mol
IFCC | Same |
| Antibody | Polyclonal anti-HbA1c from sheep blood | Same |
| Feature | Predicate Device: COBAS INTEGRA 800
Tina-quant HbA1cDx Gen.2 assay
K121291 | Candidate Device: cobas c 513 Tina-
quant HbA1cDx Gen.3 assay |
| 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 tetradecyltrimethyl-
ammonium 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 to form soluble antigen-
antibody complexes. Polyhaptens react
with excess anti-HbA1c 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. |
| 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) x 91.5 + 2.15 | Same |
| Claimed Measuring Range | Hemoglobin:
● 4-35 g/dL (2.48 – 21.7 mmol/L)
HbA1c:
● 0.3-3.4 g/dL (0.186-2.11 mmol/L) | Hemoglobin:
● 4-40 g/dL (2.48-24.8 mmol/L)
HbA1c:
● 0.3-1.93 g/dL (0.186-1.2 mmol/L) |
| Traceability | The assigned HbA1c and total hemoglobin
values of the cobas c Tina-quant
Hemoglobin A1c Gen.3 assay is certified
with the National Glycohemoglobin
Standardization Program (NGSP). The
NGSP certification expires in one year. | Same |
| Feature | Predicate Device: COBAS INTEGRA 800
Tina-quant HbA1cDx Gen.2 assay
K121291 | Candidate Device: cobas c 513 Tina-
quant HbA1cDx Gen.3 assay |
| 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 | 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: > 8µg/mL stabilizers; preservatives | 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 | 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 | sodium monohydrogenphosphate dihydrate: 64 mmol/L stabilizers; preservatives |

11

12

13

4. NON-CLINICAL PERFORMANCE EVALUATION

Performance characteristics were evaluated with the Tina-quant HbA1c Gen.3 reagent on the cobas c 513 analyzer.

This assay offers two sample type-specific applications, one for manually hemolysed samples and one for whole blood samples; thus they are named the Hemolysate and Whole Blood Applications.

The Tina-quant HbA1c Gen.3 assay first measures total hemoglobin (Hb) and glycated hemoglobin (HbA1c) in terms of mmol/L. Then the analyzer calculates the HbA1c/Hb ratio according to the corresponding protocol, 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 include limit of detection and linearity. They report results in terms of Hb and HbA1c individually. A patient sample value is reported in terms of the ratio of glycated to total hemoglobin. Method comparison, control recovery, and precision are evaluated in terms of the ratio. The protocol according to DCCT/NGSP was chosen for these studies.

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

4.1. Limit of Blank and Limit of Detection (LoB and LoD)

4.1.1. Limit of Blank

For determination of LoB one analyte free sample will be measured with three lots in 10-fold determination. Six runs distributed over > 3 days using one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot. Data analysis will be 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 (see also EP17-A2).

14

Limit of Detection 4.1.2.

For determination of LoD, five samples with low-analyte concentration will be measured with three lots in two-fold determination. Six runs distributed over > three days on one c 513 analyzer will be performed. In total, 60 measurements will be obtained per lot.

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

		Result (g/dL)	Result (mmol/L)
Limit of Blank	Hb	0.50	0.31
	HbA1c	0.19	0.12
Limit of Detection	Hb	1.00	0.62
	HbA1c	0.29	0.18

LoB and LoD Results

4.2. Precision

Precision experiments were performed in Accordance with CLSI Guideline EP5-A2 to evaluate repeatability (within-run precision) and intermediate precision of within-laboratory precision (total precision).

Two aliquots per sample were measured once each in two runs per day for 21 days on three cobas c 513 analyzers using 3 reagent lots per system. Eight total samples were evaluated in each run: two controls, PreciControl HbA1c norm and path, and 7 human samples with HbAc concentrations near 5%, 6.5%, 7.0%, 8.0%, 10.5%, 12%, and 14%.

The samples were randomized in each run separately. The data set was completed for the 21 days. 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.

15

c513 (Hemolysate) - instrument 1
Mean	Repeatability		Between-run		Between-day		Between-lot		Total
%HbA1c	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.59%	0.02	0.5	0.02	0.4	0.02	0.4	0.02	0.3	0.04	0.8
Human2.
6.18%	0.03	0.4	0.00	0.0	0.02	0.4	0.02	0.4	0.04	0.7
Human3.
6.97%	0.03	0.4	0.01	0.1	0.02	0.4	0.03	0.5	0.05	0.7
Human4,
8.05%	0.03	0.3	0.02	0.2	0.03	0.3	0.06	0.7	0.07	0.9
Human5.
10.3%	0.04	0.4	0.01	0.1	0.03	0.3	0.05	0.5	0.08	0.7
Human6.
11.8%	0.05	0.5	0.05	0.4	0.05	0.4	0.07	0.6	0.11	0.9
Human7,
13.9%	0.06	0.5	0.03	0.2	0.06	0.4	0.00	0.0	0.09	0.7
Preci Norm,
5.40%	0.02	0.4	0.01	0.1	0.02	0.4	0.01	0.2	0.03	0.6
Preci Path,
10.4%	0.04	0.4	0.00	0.0	0.04	0.4	0.06	0.6	0.08	0.8

c513 (Hemolysate) - instrument 2
Mean	Repeatability		Between-run		Between-day		Between-lot		Total
%HbA1c	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.64%	0.02	0.5	0.02	0.5	0.02	0.4	0.01	0.3	0.04	0.02
Human2,
6.25%	0.03	0.4	0.01	0.1	0.03	0.4	0.02	0.4	0.05	0.03
Human3,
7.06%	0.03	0.4	0.00	0.0	0.03	0.4	0.03	0.4	0.05	0.03
Human4,
8.14%	0.03	0.4	0.00	0.0	0.03	0.4	0.04	0.4	0.06	0.03
Human5,
10.4%	0.04	0.4	0.01	0.1	0.04	0.4	0.05	0.5	0.08	0.04
Human6,
12.0%	0.08	0.6	0.04	0.3	0.06	0.5	0.11	0.9	0.15	1.3
Human7,
13.8%	0.05	0.4	0.04	0.3	0.04	0.3	0.07	0.5	0.10	0.05
Preci Norm,
5.46%	0.03	0.5	0.01	0.2	0.02	0.4	0.02	0.4	0.04	0.03
Preci Path,
10.5%	0.04	0.4	0.02	0.2	0.04	0.4	0.05	0.5	0.08	0.04

16

c513 (Hemolysate) - instrument 3
Mean	Repeatability		Between-run		Between-day		Between-lot		Total
%HbA1c	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.67%	0.02	0.5	0.03	0.7	0.01	0.3	0.03	0.6	0.05	1.1
Human2,
6.26%	0.03	0.4	0.02	0.3	0.02	0.3	0.03	0.5	0.05	0.8
Human3,
7.06%	0.03	0.4	0.03	0.4	0.02	0.2	0.04	0.6	0.06	0.9
Human4,
8.14%	0.03	0.4	0.02	0.2	0.03	0.4	0.07	0.8	0.08	1.0
Human5.
10.4%	0.04	0.4	0.02	0.2	0.03	0.3	0.09	0.9	0.11	1.1
Human6.
11.8%	0.06	0.5	0.03	0.2	0.04	0.4	0.09	0.7	0.12	1.0
Human7,
13.8%	0.06	0.4	0.05	0.3	0.04	0.3	0.09	0.7	0.13	0.9
Preci Norm,
5.49%	0.02	0.4	0.02	0.4	0.02	0.3	0.03	0.5	0.04	0.8
Preci Path.
10.5%	0.04	0.4	0.03	0.2	0.03	0.3	0.09	0.9	0.11	1.1

c513 Hemolysate - all 3 instruments
Mean
%HbA1c	Repeatability		Between
Run		Between
Day		Between
Lot		Between
Instrument		Total
	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.63%	0.02	0.5	0.03	0.6	0.02	0.4	0.02	0.4	0.04	0.9	0.06	1.3
Human2,
6.23%	0.03	0.4	0.01	0.2	0.02	0.4	0.03	0.4	0.04	0.7	0.06	1.0
Human3,
7.03%	0.03	0.4	0.02	0.2	0.02	0.3	0.04	0.5	0.05	0.7	0.07	1.0
Human4,
8.11%	0.03	0.4	0.01	0.2	0.03	0.4	0.06	0.7	0.04	0.5	0.08	1.0
Human5,
10.4%	0.04	0.4	0.02	0.2	0.04	0.3	0.07	0.7	0.03	0.3	0.10	0.9
Human6,
11.9%	0.06	0.5	0.04	0.3	0.05	0.5	0.09	0.7	0.09	0.7	0.15	1.3
Human7,
13.8%	0.06	0.4	0.04	0.3	0.05	0.4	0.07	0.5	0.02	0.1	0.11	0.8
Preci
Norm
5.45%	0.02	0.4	0.01	0.3	0.02	0.4	0.02	0.4	0.04	0.8	0.06	1.1
Preci Path,
10.4%	0.04	0.4	0.01	0.1	0.04	0.4	0.07	0.7	0.03	0.3	0.10	0.9

17

c513 (Whole Blood) - instrument 1
Mean
%HbA1c	Repeatability		Between-run		Between-day		Between-lot		Total
	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.54%	0.03	0.6	0.02	0.5	0.01	0.3	0.07	1.5	0.08	1.7
Human2,
6.21%	0.02	0.4	0.01	0.1	0.02	0.3	0.01	0.2	0.03	0.6
Human3,
6.97%	0.03	0.4	0.01	0.2	0.02	0.3	0.01	0.2	0.04	0.6
Human4,
8.10%	0.04	0.5	0.01	0.1	0.03	0.4	0.04	0.5	0.06	0.8
Human5,
10.5%	0.04	0.4	0.04	0.4	0.01	0.1	0.09	0.8	0.10	1.0
Human6,
11.8%	0.05	0.4	0.04	0.4	0.08	0.6	0.10	0.8	0.14	1.2
Human7,
13.9%	0.06	0.5	0.06	0.4	0.05	0.4	0.06	0.4	0.12	0.8
Preci Norm,
5.42%	0.03	0.5	0.01	0.2	0.02	0.4	0.03	0.5	0.05	0.8
Preci Path,
10.6%	0.04	0.4	0.04	0.4	0.03	0.3	0.08	0.8	0.10	1.0

c513 (Whole Blood) - instrument 2
Mean	Repeatability		Between-run		Between-day		Between-lot		Total
%HbA1c	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.64%	0.03	0.6	0.03	0.7	0.01	0.2	0.02	0.4	0.05	1.0
Human2,
6.29%	0.03	0.4	0.01	0.2	0.02	0.3	0.02	0.3	0.04	0.6
Human3,
7.05%	0.03	0.5	0.03	0.5	0.01	0.1	0.02	0.2	0.05	0.7
Human4,
8.17%	0.04	0.5	0.03	0.4	0.02	0.3	0.05	0.6	0.07	0.9
Human5.
10.5%	0.05	0.5	0.03	0.3	0.02	0.2	0.07	0.6	0.09	0.8
Human6,
12.1%	0.07	0.6	0.04	0.3	0.07	0.6	0.15	1.3	0.19	1.5
Human7,
13.9%	0.06	0.4	0.05	0.3	0.08	0.6	0.13	0.9	0.17	1.2
Preci Norm,
5.51%	0.03	0.5	0.02	0.4	0.01	0.2	0.01	0.2	0.04	0.7
Preci Path,
10.6%	0.05	0.5	0.04	0.4	0.01	0.1	0.07	0.6	0.09	0.9

18

c513 (Whole Blood) - instrument3
Mean
%HbA1c	Repeatability		Between-run		Between-day		Between-lot		Total
	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.61%	0.03	0.7	0.02	0.4	0.02	0.3	0.01	0.2	0.04	0.9
Human2,
6.27%	0.03	0.4	0.02	0.3	0.01	0.2	0.04	0.6	0.05	0.8
Human3,
7.04%	0.04	0.5	0.02	0.3	0.01	0.2	0.04	0.6	0.06	0.9
Human4,
8.21%	0.04	0.5	0.04	0.4	0.01	0.1	0.09	1.1	0.10	1.2
Human5,
10.6%	0.04	0.4	0.04	0.4	0.00	0.0	0.13	1.2	0.14	1.3
Human6,
12.0%	0.06	0.5	0.05	0.4	0.05	0.4	0.07	0.6	0.11	0.9
Human7,
13.8%	0.07	0.5	0.03	0.2	0.04	0.3	0.27	2.0	0.28	2.1
Preci Norm,
5.48%	0.03	0.5	0.01	0.2	0.01	0.3	0.01	0.3	0.04	0.7
Preci Path,
10.7%	0.05	0.5	0.06	0.6	0.00	0.0	0.13	1.2	0.15	1.4

c513 - whole blood - all 3 instruments
Mean
%HbA1c	Repeatability		Between
Run		Between
Day		Between Lot		Between
Instrument		Total
	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Human1,
4.60%	0.03	0.6	0.02	0.5	0.01	0.3	0.04	0.9	0.04	1.0	0.07	1.6
Human2,
6.26%	0.03	0.4	0.01	0.2	0.02	0.3	0.02	0.4	0.04	0.6	0.06	0.9
Human3,
7.02%	0.03	0.5	0.02	0.4	0.02	0.2	0.03	0.4	0.04	0.5	0.06	0.9
Human4,
8.16%	0.04	0.5	0.03	0.3	0.02	0.3	0.06	0.8	0.04	0.5	0.09	1.1
Human5,
10.5%	0.04	0.4	0.04	0.4	0.01	0.1	0.10	0.9	0.00	0.0	0.11	1.1
Human6,
12.0%	0.06	0.5	0.04	0.3	0.07	0.6	0.11	0.9	0.11	0.9	0.19	1.5
Human7,
13.9%	0.06	0.5	0.05	0.3	0.06	0.4	0.18	1.3	0.00	0.0	0.20	1.5
PreciNorm
5.47%	0.03	0.5	0.01	0.3	0.02	0.3	0.02	0.3	0.04	0.8	0.06	1.1
PreciPath,
10.6%	0.05	0.5	0.05	0.5	0.01	0.1	0.09	0.9	0.00	0.0	0.12	1.1

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4.3. Method Comparison

A method comparison study was performed to compare the sample results from the candidate method, Tina-quant HbA1c Gen.3 on the cobas c 513, to results from the TOSOH 8 HPLC. This study will be conducted with both the Tina-quant HbA1c Gen.3 Whole Blood Application and Hemolysate Application.

One hundred and fifty-five (155) samples from the secondary NGSP reference laboratory were used in the evaluation. These samples were measured by the secondary NGSP reference laboratory using a Tosoh HPLC system (X axis) and by the Roche Tina-quant HbA1c Gen.3 test system (Y axis). Samples were tested over a 3 day period with one lot of reagent on one cobas c 513 analyzer.

All acceptance criteria for method comparison were met. The difference plots show there is good agreement between the Roche method and the NGSP TOSOH HPLC reference method. The table below summarizes the bias between the Tina-quant HbA1c Gen.3 (Whole Blood and Hemolysate applications) and the NGSP's TOSOH HPLC reference method.

| | Tina-quant HbA1c Gen.3
Whole Blood Application | Tina-quant HbA1c Gen.3
Hemolysate Application |
|---------------------------|---------------------------------------------------|--------------------------------------------------|
| Mean bias vs. NGSP TOSOH | -0.016 | -0.138 |
| Mean bias at lower 95% CI | -0.260 | -0.429 |
| Mean bias at upper 95% CI | 0.227 | 0.153 |

Difference Plot Analysis Data Summary

The table below summarizes the percent relative bias at various points in the measuring range for both applications.

| %HbA1c | Bias limit | % Relative Bias
Whole Blood Application | % Relative Bias
Hemolysate Application |
|--------|------------|--------------------------------------------|-------------------------------------------|
| >5% | ≤ 5.0% | 0.1% | -0.8% |
| 6.0% | ≤ 3.5% | -0.2% | -1.4% |
| 6.5% | ≤ 3.5% | -0.3% | -1.6% |
| 7.0% | ≤ 3.5% | -0.4% | -1.8% |
| >12.0% | ≤ 10% | -0.8% | -2.8% |

Bias at Concentration Data Summary

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Method Comparison Sample Distribution

Hemolysate
%
HbA1c	Samples
Tested	% of Total
≤ 5%	6	3.9%
5 – 6%	25	16.1%
6 – 6.5%	30	19.4%
6.5 – 7%	36	23.2%
7 – 8%	25	16.1%
8 – 9 %	14	9.0%
> 9%	19	12.3%
Total	155	100.0%

Whole blood
%	Samples	% of
HbA1c	Tested	Total
≤ 5%	6	3.9%
5 - 6%	26	16.9%
6 - 6.5%	30	19.5%
6.5 – 7%	36	23.4%
7-8%	25	16.2%
8-9%	14	9.1%
> 9%	17	11.0%
Total	154	100.0%

4.4. Total Error Near the Cut-off

Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the precision study, the Total Error (TE) at the following concentrations (5%, 6.5%, 8% and 12%) was calculated as follows: %TE =%Bias| + 1.96 * %CV * (1+%Bias). The results are presented in the tables below.

Total Error - Whole Blood Application

% HbA1c	%BIAS	%CV	Total Error (%)
5.0% HbA1c	0.7	1.6	3.9
6.5% HbA1c	0.3	0.9	2.1
8.0% HbA1c	0.1	1.1	2.3
12.0% HbA1c	0.2	1.5	3.1

Total Error – Hemolysate Application

% HbA1c	%BIAS	%CV	Total Error (%)
5.0% HbA1c	1.1	1.3	3.7
6.5% HbA1c	1.7	1.0	3.7
8.0% HbA1c	2.0	1.0	4.0
12.0% HbA1c	2.6	1.3	5.2

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4.5. Linearity

Two separate dilution series consisting of at least 11 levels were prepared using the human hemolysate sample pools with either HbA1c or Hb with 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 done separately for each sample. Linear regression analysis was done according to EP6-A. Additionally, first order regression statistics using theoretical values determined from sample pools with known concentrations is provided in the table below.

| Application | Analyte | Low End of Linear
Range | | High End of Linear
Range | | Slope | Intercept | Pearson's
r |
|-------------|---------|----------------------------|--------|-----------------------------|--------|-------|-----------|----------------|
| | | g/dL | mmol/L | g/dL | mmol/L | | | |
| Hemolysate | Hb | 3.4 | 2.09 | 40.3 | 25.0 | 0.983 | 0.051 | 0.9999 |
| | HbA1c | 0.28 | 0.177 | 2.85 | 1.77 | 0.972 | 0.002 | 0.9990 |

Empirical First Order Linear Regression Results

Claimed Ranges:

Hemoglobin: 4.0 - 40 g/dL (2.48-24.8 mmol/L)

HbA1c: 0.3 - 1.93 g/dL (0.186-1.20 mmol/L)

This corresponds to a measuring range of 23-146 mmol/mol HbA1c (IFCC) and 4.2-15.5 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L.

4.6. Matrix Comparison

A matrix comparison study was performed to evaluate different anticoagulants on analyte recovery. At least 40 samples of each sample type and at least 40 half-filled tubes of each sample type were evaluated. The half-filled (double concentrated) and filled sample tubes were from one donor. Only samples within the limit of icteric, lipemic and hemolytic interference

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were used. Samples covered the measuring range. The Hemolysate application was used for these measurements. All acceptance criteria for matrix comparison were met.

| Sample Type | Anticoagulant | Tube Fill | Mean
Difference | Upper 95% | Lower 95% |
|-------------|---------------|-----------|--------------------|-----------|-----------|
| Hemolysate | K2-EDTA | ½ full | -0.0559 | 0.319 | -0.431 |
| | K3-EDTA | Full | 0.000732 | 0.418 | -0.417 |
| | K3-EDTA | ½ full | -0.0351 | 0.506 | -0.576 |
| | Li Heparin | Full | 0.0156 | 0.501 | -0.470 |
| | Li Heparin | ½ full | -0.0285 | 0.504 | -0.561 |
| | NaF/Potox | Full | -0.00732 | 0.459 | -0.473 |
| NaF/Potox | ½ full | 0.0393 | 0.483 | -0.404 | |

Matrix Comparison Results

Endogenous Interference 4.7.

A study evaluated several endogenous substances for potential interference with the measure of % HbA1c. The following six endogenous substances were evaluated:

Lipemia .
Bilirubin ●
. Ditaurobilirubin
. Glucose
. Rheumatoid Factor
Total Protein ●

Pooled whole blood samples with two A1c levels, one near the medical decision level and one above it, were spiked with the maximum level of the above six interferents in separate preparations resulting in 12 spiked samples. These samples were then hemolyzed with Tinaquant HbA1c Hemolyzing Reagent. Another pool, without interferent, was equally hemolyzed. A minimum of a 10-level dilution series was then created for each of the 12 spiked samples by using the interferent free pool as the diluting reagent.

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The experiment was performed with one reagent lot, one c 513 analyzer, and in a single run from the same calibration. PreciControl HbA1c norm and path will be the controls. The twelve dilution series were tested ten-fold for % HbA1c using the Hemolysate Application only. Interference is a matter of specificity of the assay which is independent of the application; therefore, interference data on the Hemolysate application will be representative of data on both the Hemolysate and Whole Blood Applications.

The mean of the ten replicates was determined and compared to the result from the reference sample (aliquot with no interfering substance). The comparison was evaluated as a percent deviation. An interferent will be significant if it causes >7% deviation of a measurement in terms of % HbA1c.

Endogenous Interference

| Interferent | Maximum Concentration without interference
Claimed |
|-------------------|-------------------------------------------------------|
| Lipemia | 600 mg/dL |
| Bilirubin | 60 mg/dL |
| Ditaurobilirubin | 60 mg/dL |
| Glucose | 1000 mg/dL |
| Rheumatoid Factor | 750 IU/mL |

4.8. Drug Interferences

A study evaluated several drugs for potential interference with the measurement of %HbA1c using the hemolysate application. The following drugs were studied:

. N- Acetylcysteine
Ampicillin-Na .
Ascorbic acid .
Cefoxitin ●
Heparin ●
Levodopa ●
. Methyldopa

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. Metronidazole
Doxycyclin
Acetylsalicylic acid ●
Rifampicin ●
Cyclosporine ●
Phenylbutazone ●
Acetaminophen ●
Ibuprofen .
. Theophylline

The 16 commonly used drugs listed above were added to native patient samples and examined for potential effect on % HbA1c determination with the Hemolysate Application. Drug interference testing was performed with hemolysate samples at 2 different HbA1c levels. The two different HbA1c concentrations will be approximately 6% and 8% HbA1c. Each drug was added in two defined concentrations with concentration 1 being several times (typically 5 times) the maximum daily dosage and concentration 2 being the maximum daily dosage level. Concentration 1 was performed for screening purposes only and concentration 2 is the relevant drug concentration for determining interferences with the assay. Samples were measured in tenfold using the cobas c 513 system. The median value was compared to the reference value (HbA1c sample with no drug added) and the deviation from the reference was calculated.

The data was collected using the Hemolysate Application which is acceptable since the assay interference is a matter of specificity of the antibody.

Significant interference is defined as > ±7% deviation from the reference value observed. All acceptance criteria for drug interferences were met.

Cross reactivity 4.9.

Studies were performed to determine if the Tina-quant Hemoglobin A1cDx Gen.3 assay demonstrates cross-reactivity with any of the following hemoglobin fractions and glycated albumin.

25

HbA0 ●
HbA1a+b ●
Acetylated Hb ●
Carbamylated Hb ●
Glycated Albumin .
Labile HbA1c ●

A series experiments were performed with one reagent lot, one c 513 analyzer, and in a single run from the same calibration. PreciControl HbA1c norm and path function as the controls. Ten replicates of each sample were analyzed. The % recovery of the median for each dilution step related to the median obtained for Pool 1 will be plotted against the dilution step and/or the concentration of the potential cross-reactant.

As with the interference studies, the data was collected using the Hemolysate Application only. This is acceptable because cross-reactivity is a matter of specificity of the assay which is independent of the application. Therefore, cross-reactivity data on the Hemolysate Application are representative of data on both the Hemolysate and Whole Blood Application.

| Cross-Reactant | | Max Concentration
Measured | Max Concentration
with no interference |
|------------------|---------|-------------------------------|-------------------------------------------|
| HbA0 | Level 1 | 120 g/dL | 120 g/dL |
| | Level 2 | 120 g/dL | 120 g/dL |
| HbA1a+b | Level 1 | 1.60 g/dL | 0.96 g/dL |
| | Level 2 | 1.60 g/dL | 1.60 g/dL |
| Acetylated Hb | Level 1 | 2.00 g/dL | 2.00 g/dL |
| | Level 2 | 2.00 g/dL | 2.00 g/dL |
| Carbamylated Hb | Level 1 | 2.0 g/dL | 2.0 g/dL |
| | Level 2 | 2.0 g/dL | 2.0 g/dL |
| Glycated albumin | Level 1 | 10.0 g/dL | 10.0 g/dL |
| | Level 2 | 10.0 g/dL | 10.0 g/dL |
| Labile HbA1c | Level 1 | 1500 mg/dL | 1500 mg/dL |
| | Level 2 | 1500 mg/dL | 1500 mg/dL |

Cross-Reactivity

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4.10. Hemoglobin Variants

Hemoglobin variant testing was conducted to determine if there was any significant interference with any of the major hemoglobin variants and the Tina-quant Hemoglobin A1cDx Gen.3 assay. 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. Moreover, in some conditions, the fetal hemoglobin HbF is elevated. Also, the erythrocytes of some patients (e.g. beta thalassemia minor) contain elevated levels of HbA2. 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	20	31 - 42% S	5.0-14.4
HbC	20	33 - 44% C	4.7-13.0
HbE	20	27 - 33% E	5.0-9.7
HbD	20	34 - 42% D	5.0-10.9
Elevated F	20	2 - 28% F	5.8-10.1
HbA2	13	4.3 - 6.2% A2	5-10

Hemoglobin Variant Samples

Each sample was tested once in at least one run on one c 513 analyzer.

Results obtained with the Tina-quant Hemoglobin A1cDx Gen.3 assay (Hemolysate Application) on the c 513 analyzer will be compared to those obtained with the reference methods.

For purposes of this experiment, the data was collected using the Hemolysate Application. This is acceptable since variant interference is a matter of specificity of the antibody. Therefore, the data shown is representative of both the whole blood and hemolysate applications.

There was no significance interference with any of the major hemoglobin variants. Results are summarized in the table below.

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Percent Relative Bias from Reference Method at Low and High Concentrations of HbA1c Samples
	~6.0% HbA1c		~9.0% HbA1c
Hb Variant	Relative %
Difference	Range	Relative %
Difference	Range
HbA2	-2.0%	-3.5 - (-1.3)	1.2%	-1.2 - 2.5
HbC	-1.3%	-4.0 - 1.79	-1.3%	-4.79 - 2.45
HbD	-1.5%	-1.9 - 0.8	-2.3%	-2.4 - (-0.3)
HbE	-1.8%	-4.5 - (-0.2)	0.7%	-0.9 - 2.5
HbS	-1.6%	-4.1 - 3.4	2.7%	-0.3 - 3.0
HbF	Specimens containing high amounts of HbF (> 7 %) may yield lower than expected HbA1c
values.

Hemoglobin Variant Testing Results

Heterozygous presence of the most common hemoglobin variants (HbAS, HbAD, HbAE) does not interfere. Significant interference was defined as ≥7% change in HbA1c value in the presence of the hemoglobin variant relative to control.

5. CONCLUSIONS

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