CAPI 3 Hb A1c

K171537 · Sebia · PDJ · Sep 12, 2017 · Clinical Chemistry

Device Facts

• Record ID: K171537
• Device Name: CAPI 3 Hb A1c
• Applicant: Sebia
• Product Code: PDJ · Clinical Chemistry
• Decision Date: Sep 12, 2017
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1373
• Device Class: Class 2

Intended Use

The CAPI 3 Hb A1c kit is intended for separation and quantification of the HbA1c glycated fraction of hemoglobin (in IFCC unit (mmol/mol) and NGSP unit (%)) in venous whole human blood, by capillary electrophoresis in alkaline buffer with the CAPILLARYS 3 TERA instrument. Measurement of hemoglobin A1c is used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus. The CAPI 3 Hb A1c kit is intended for in vitro Diagnostic Use Only.

Device Story

The CAPI 3 Hb A1c kit is an in vitro diagnostic assay used with the CAPILLARYS 3 TERA instrument for the separation and quantification of HbA1c in venous whole human blood. The system utilizes capillary electrophoresis in free solution; samples are diluted with hemolysing solution and injected into silica capillaries. Charged hemoglobin molecules are separated based on electrophoretic mobility in an alkaline buffer. Detection occurs at the cathodic end of the capillary at 415 nm. The instrument processes 12 samples simultaneously. The output provides HbA1c concentrations in NGSP (%) and IFCC (mmol/mol) units. Results are used by clinicians to diagnose diabetes, identify at-risk patients, and monitor long-term glycemic control. The device provides high-resolution separation, allowing quantification in the presence of labile HbA1c, carbamylated/acetylated hemoglobins, and major hemoglobin variants (S, C, E, D, F).

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP05-A3 using 4 concentrations (~5%, 6.5%, 8%, 12%) over 24 days, 1728 results. Linearity confirmed across 4.0-16.5% (NGSP) range. Method comparison with 152 variant-free samples against a reference HPLC method showed r=0.999, slope=1.014, and y-intercept=-0.142. Total error was ≤6% across all tested concentrations. Interference studies confirmed no significant interference from common drugs, endogenous factors, or hemoglobin variants (S, C, D, E, A2), except for HbF >23%.

Technological Characteristics

Capillary electrophoresis in free solution; silica capillaries; alkaline buffer; 415 nm absorbance detection. Measures HbA1c in venous whole blood. Standardized to IFCC and NGSP. Connectivity: CAPILLARYS 3 TERA instrument. Software-based analysis of electrophoretic peaks.

Indications for Use

Indicated for individuals with diabetes mellitus or those at risk for developing diabetes mellitus. Used for diagnosis and monitoring of long-term blood glucose control. Contraindicated for patients with fetal hemoglobin (HbF) concentrations >23%.

Regulatory Classification

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.

Special Controls

(b) Classification. Class II (special controls). Hemoglobin A1c test systems must comply with the following special controls: 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%, 6.5%, 8.0% and 12% hemoglobin A1c. This testing must evaluate precision over a minimum of 20 days using at least 3 lots of the device and 3 instruments, as applicable. ii) Performance testing of device accuracy must include a minimum of 120 blood samples that span the measuring interval of the new device and compare results of the new device to results of the 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%. 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.

*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.

Predicate Devices

• Tosoh Automated Glycohemoglobin Analyzer HLC-723G-8 (k131580)

Submission Summary (Full Text)

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

September 12, 2017

SEBIA, INC. KAREN ANDERSON DIRECTOR OF TECHNICAL AND QUALITY ASSURANCE 1705 CORPORATE DRIVE SUITE 400 NORCROSS GA 30093

Re: k171537

Trade/Device Name: CAPI 3 Hb A1c Regulation Number: 21 CFR 862.1373 Regulation Name: Hemoglobin A1c Test System Regulatory Class: II Product Code: PDJ Dated: July 24, 2017 Received: July 27, 2017

Dear Ms. Karen Anderson:

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 (QS) 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 [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/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 [phone] or [phone] or at its Internet address

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

Sincerely yours.

Courtney H. Lias -S

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) k171537

Device Name CAPI 3 Hb A1c

#### Indications for Use (Describe)

The CAPI 3 Hb A1c kit is intended for separation and quantification of the HbA1c glycated fraction of hemoglobin (in IFCC unit (mmol/mol) and NGSP unit (%)) in venous whole human blood, by capillary electrophoresis in alkaline buffer with the CAPILLARYS 3 TERA instrument of hemoglobin A1c is used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus. The CAPI 3 Hb A1c kit is intended for in vitro Diagnostic Use Only.

| Type of Use (Select one or both, as applicable) |
|-------------------------------------------------------------------------------------------------------------------------------|
| <div> <span> <span style="text-decoration: overline;">X</span> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> |
| <div> <span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> </div> |

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## 510K SUMMARY (Summary of Safety and Effectiveness) - K171537

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

| Submitter Name | Sebia, Inc. |
|---------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Address | 1705 Corporate Drive Suite 400<br>Norcross, Georgia 30093, USA |
| Contact | Karen Anderson, Dir of Technical and QA<br>Phone: [phone]<br>Fax: [phone]<br>Email: [e-mail]<br>Aigars Brants, Ph.D, Scientific Affairs Officer<br>Phone [phone]<br>Fax [phone]<br>Email: [e-mail] |
| Date Prepared | August 14, 2017 |
| Manufacturing | Sebia<br>Parc Technologique Léonard de Vinci<br>Rue Léonard de Vinci,<br>CP 8010 LISSES, 91008 EVRY Cedex<br>FRANCE<br>Phone: [phone]<br>Fax: [phone] |
| Product Name | CAPI 3 Hb A1c |
| Common Name | Whole blood hemoglobin A1c (HbA1c) by capillary<br>electrophoresis |
| Product Regulation No. | 21 CFR 862.1373 |
| Product Codes | PDJ |
| Device classification and Panel<br>Classification | Class II , Clinical Chemistry(75) |
| Establishment Registration No. | 8023024 |

510(k) Summary 1

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## 1. DEVICE DESCRIPTION

The CAPILLARYS 3 TERA instrument uses the principle of capillary electrophoresis in free solution. With this technique, charged molecules are separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation occurs according to the electrolyte pH and electroosmotic flow.

The CAPILLARYS 3 TERA instrument has silica capillaries functioning in parallel allowing 12 simultaneous analyses of HbA1c quantification in a whole blood sample. A sample dilution with hemolysing solution is prepared and injected by aspiration at the anodic end of the capillary. A high voltage protein separation is then performed and direct detection of the hemoglobins is made at the cathodic end of the capillary at 415 nm, which is the absorbance wave length specific to hemoglobins. Before each run, the capillaries are washed with a wash solution and prepared for the next analysis with buffer.

Direct detection provides accurate relative quantification of individual hemoglobin A1c fraction. In addition, the high resolution of CAPI 3 Hb A1c procedure allows the quantification of HbA1c even in the presence of labile HbA1c. carbamylated and acetylated hemoglobins, and major hemoqlobin variants.

By using an alkaline pH buffer, normal and abnormal (or variant) hemoglobins are detected in the following order, from cathode to anode: A2/C, E, S/D, F, A0, other Hb (including minor Hb A1) and then A1c.

The HbA1c concentrations are standardized and indicated in %HbA1c (DCCT/NGSP) and in mmol/mol (IFCC) units.

### Reagents:

### CAPI 3 Hb A1c KIT

| ITEMS | PN 2515 |
|------------------------------------|----------------------|
| Buffer (ready to use) | 2 vials, 700 mL each |
| Hemolysing solution (ready to use) | 1 vial, 700 mL |
| Filters | 4 filters |

### Additional reagents not included in the CAPI 3 Hb A1c KIT

| ITEMS | PN | COMPONENTS |
|----------------------------|------|---------------|
| CAPICLEAN CAPILLARYS 3 | 2060 | 1 vial, 25 mL |
| CAPILLARYS 3 WASH SOLUTION | 2062 | 1 vial, 75mL |

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| CAPI 3 DISPOSABLES KIT | 2580 | 10 packs of 14 reagent cups<br>5 bins for used reagent cups |
|--------------------------------------|------|-------------------------------------------------------------|
| TEST TUBES | 9214 | 200 of 100 mm-tubes |
| CAPI 3 BINS FOR USED REAGENT<br>CUPS | 2581 | 5 units |
| TUBES AND CAPS FOR | 9202 | 20 units |
| CONTROLS | 9205 | 500 units |
| CAPI 3 REAGENT CUPS | 2582 | 24 packs of 14 reagent cups |

## 2. INDICATIONS FOR USE

## CAPI 3 Hb A1c kit:

The CAPI 3 Hb A1c kit is intended for separation and quantification of the HbA1c glycated fraction of hemoglobin (in IFCC unit (mmol/mol) and NGSP unit (%)) in venous whole human blood, by capillary electrophoresis in alkaline buffer with the CAPILLARYS 3 TERA instrument. Measurement of hemoglobin A1c is used as an aid in diagnosis of diabetes, as an aid to identify patients who may be at risk for developing diabetes mellitus, and for the monitoring of longterm blood glucose control in individuals with diabetes mellitus. The CAPI 3 Hb A1c kit is intended for in vitro Diagnostic Use Only.

## 3. TECHNOLOGICAL CHARACTERISTICS

The CAPILLARYS 3 TERA instrument uses the principle of capillary electrophoresis in free solution which is the most common form of capillary electrophoresis. With this technique, charged molecules are separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation also occurs according to the electrolyte pH and electroosmotic flow.

The CAPILLARYS 3 TERA instrument has silica capillaries functioning in parallel allowing 12 simultaneous analyses of Hb A1c quantification in a whole blood sample. A sample dilution with hemolysing solution is prepared and injected by aspiration at the anodic end of the capillary. A high voltage protein separation is then performed and direct detection of the hemoglobins is made at the cathodic end of the capillary at 415 nm, which is the absorbance wave length specific to hemoglobins. Before each run, the capillaries are washed with a wash solution and prepared for the next analysis with buffer.

Direct detection provides accurate relative quantification of individual hemoglobin A1cfraction. In addition, the high resolution of CAPI 3 Hb A1c procedure allows the quantification of HbA1c, and particularly, even in the presence of labile HbA1c, carbamylated and acetylated hemoglobins, and major hemoglobin variants.

By using an alkaline pH buffer, normal and abnormal (or variant) hemoglobins are detected in the following order, from cathode to anode: A2/C, E, S/D, F, A0, other Hb (including minor Hb A1) and then A1c.

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## 4. SUBSTANTIAL EQUIVALENCE INFORMATION:

| Predicate Device Name | Predicate Device<br>510(k) number | Product Code | Regulation No |
|------------------------------------------------------------|-----------------------------------|--------------|---------------|
| Tosoh Automated<br>Glycohemoglobin Analyzer HLC-<br>723G-8 | k131580 | PDJ | 862.1373 |

Similarities between the candidate device (CAPI 3 Hb A1c) and the predicate device (Tosoh HLC-723G8, k131580 (Table A).

| | Similarities | |
|------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Table A | Sebia CAPI 3 Hb A1c Candidate<br>Device | Tosoh HLC-723G8 Predicate<br>Device (k131580) |
| Intended use | The CAPI 3 Hb A1c kit is intended for<br>separation and quantification of the HbA1c<br>glycated fraction of hemoglobin (in IFCC unit<br>(mmol/mol) and NGSP unit (%)) in venous<br>whole human blood, by capillary<br>electrophoresis in alkaline buffer with the<br>CAPILLARYS 3 TERA instrument.<br>Measurement of hemoglobin A1c is used as<br>an aid in diagnosis of diabetes, as an aid to<br>identify patients who may be at risk for<br>developing diabetes mellitus, and for the<br>monitoring of long-term blood glucose control<br>in individuals with diabetes mellitus. The CAPI<br>3 Hb A1c kit is intended for in vitro Diagnostic<br>Use Only. | The Tosoh Automated<br>Glycohemoglobin Analyzer<br>HLC723G8 is intended for the in vitro<br>diagnostic use for the measurement of<br>% hemoglobin A1c (HbA1c)<br>(DCCT/NGSP) and mmol/mol<br>hemoglobin A1c (IFCC) in whole<br>blood specimens. This test is to be<br>used as an aid in diagnosis of<br>diabetes and identifying patients who<br>may be at risk of developing diabetes. |
| Specimen Type | Human Whole Blood | Human Whole Blood |
| Standardization | Traceable to the Diabetes Control and<br>Complications Trial (DCCT) reference<br>method and IFCC. Certified via the National<br>Glycohemoglobin Standardization Program (<br>NGSP) | Same |
| Linearity<br>Measuring Range | 4.0 - 16.5% (NGSP)<br>20 - 157 mmol/mol (IFCC) | 4.0 - 16.9 % |

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| | Concentration | TE% | Concentration | TE% |
|----------------------------------------|-----------------------------------------------------------------|-----|---------------|-----------------------------------------------------------------|
| Total Error<br>allowable bias 6% | 5.2 | 4.4 | 5.0 | 5.8 |
| | 6.5 | 2.9 | 6.5 | 2.8 |
| | 8.1 | 2.3 | 8.0 | 3.0 |
| | 11.9 | 3.6 | 12.0 | 3.1 |
| Hemoglobin<br>Variant<br>Interferences | HbA2, HbF, HbS, HbC, HbD,<br>does not interfere with this assay | | | HbA2, HbF, HbS, HbC, HbD, does<br>not interfere with this assay |

Table B. Differences between the predicate device (CAPI 3 Hb A1c) and the candidate device (Tosoh HLC-723G8, k131580) in (Table B).

| | Differences | |
|----------------------------------------|-----------------------------------------|---------------------------------------------------------------------------------------|
| Table B | Sebia CAPI 3 Hb A1c<br>Candidate Device | Tosoh HLC-723G8<br>Predicate Device (k131580) |
| Assay Principle | Capillary electrophoresis | Ion-exchange HPLC |
| Hemoglobin<br>Variant<br>Interferences | Hb E no interference | Hb E has known interference, a HbE<br>flag is displayed and no Hb A1c is<br>reported. |

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### 5. Performance Data:

### a. Precision / Reproducibility:-

The precision of the CAPI 3 Hb A1c procedure using the CAPILLARYS 3 TERA was evaluated based on CLSI guidelines EP05-A3 guidelines, Evaluation of Quantitative Measurement Procedures. Four whole blood samples at the following targeted HbA1c concentration of ~ 5%, ~ 6.5%, ~ 8%, and ~12% were used in the study. The study included two quality control materials and two calibrators. The samples were analyzed in duplicate on two capillaries per run on 3 instruments. The studied used three lots of kits over 24 days yielding a total of 1728 results over total testing time of 72 days.

| Sample | Mean<br>(mmol/mol) | Within-<br>capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Betweeninstrument | | Total<br>reproducibility<br>(*) | |
|-----------------|--------------------|----------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|-------------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No.<br>1 | 33 | 0,50 | 1,5% | 0,48 | 1,4% | 0,00 | 0,0% | 0,24 | 0,7% | 0,40 | 1,2% | 0,18 | 0,6% | 0,86 | 2,6% |
| Sample No.<br>2 | 34 | 0,49 | 1,4% | 0,52 | 1,5% | 0,00 | 0,0% | 0,28 | 0,8% | 0,49 | 1,5% | 0,00 | 0,0% | 0,91 | 2,7% |
| Sample No.<br>3 | 37 | 0,44 | 1,2% | 0,51 | 1,4% | 0,00 | 0,0% | 0,27 | 0,7% | 0,42 | 1,1% | 0,00 | 0,0% | 0,84 | 2,3% |
| Sample No.<br>4 | 48 | 0,56 | 1,2% | 0,43 | 0,9% | 0,26 | 0,5% | 0,22 | 0,5% | 0,38 | 0,8% | 0,00 | 0,0% | 0,83 | 1,7% |
| Sample No.<br>5 | 61 | 0,53 | 0,9% | 0,43 | 0,7% | 0,00 | 0,0% | 0,43 | 0,7% | 0,09 | 0,1% | 1,52 | 2,6% | 1,72 | 2,9% |
| Sample No.<br>6 | 65 | 0,62 | 1,0% | 0,43 | 0,7% | 0,00 | 0,0% | 0,31 | 0,5% | 0,23 | 0,4% | 0,13 | 0,2% | 0,86 | 1,3% |
| Sample No.<br>7 | 86 | 0,64 | 0,7% | 0,49 | 0,6% | 0,00 | 0,0% | 0,39 | 0,5% | 0,72 | 0,8% | 0,00 | 0,0% | 1,14 | 1,3% |
| Sample No.<br>8 | 107 | 0,74 | 0,7% | 0,83 | 0,8% | 0,00 | 0,0% | 0,67 | 0,6% | 1,79 | 1,7% | 0,00 | 0,0% | 2,21 | 2,1% |

(*) Total reproducibility includes : within-capillary, between-run, between-day, between-lot and between-instrument.

| Sample | Mean | Within-<br>capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Betweeninstrument | | Total<br>reproducibility<br>(*) | |
|-----------------|------|----------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|-------------------|------|---------------------------------|------|
| | (%) | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No.<br>1 | 5,2 | 0,05 | 0,9% | 0,04 | 0,8% | 0,00 | 0,0% | 0,02 | 0,4% | 0,04 | 0,7% | 0,02 | 0,3% | 0,08 | 1,5% |
| Sample No.<br>2 | 5,2 | 0,05 | 0,9% | 0,05 | 0,9% | 0,00 | 0,0% | 0,03 | 0,6% | 0,04 | 0,7% | 0,00 | 0,0% | 0,08 | 1,6% |
| Sample No.<br>3 | 5,5 | 0,04 | 0,8% | 0,04 | 0,8% | 0,00 | 0,0% | 0,03 | 0,5% | 0,04 | 0,7% | 0,00 | 0,0% | 0,08 | 1,4% |
| Sample No.<br>4 | 6,5 | 0,05 | 0,7% | 0,04 | 0,5% | 0,02 | 0,3% | 0,02 | 0,3% | 0,03 | 0,5% | 0,00 | 0,0% | 0,07 | 1,1% |
| Sample No.<br>5 | 7,7 | 0,05 | 0,7% | 0,04 | 0,5% | 0,00 | 0,0% | 0,04 | 0,5% | 0,01 | 0,2% | 0,13 | 1,7% | 0,15 | 2,0% |
| Sample No.<br>6 | 8,1 | 0,06 | 0,7% | 0,04 | 0,5% | 0,00 | 0,0% | 0,03 | 0,4% | 0,02 | 0,3% | 0,01 | 0,1% | 0,08 | 1,0% |
| Sample No.<br>7 | 10,1 | 0,06 | 0,6% | 0,05 | 0,5% | 0,00 | 0,0% | 0,04 | 0,4% | 0,07 | 0,7% | 0,00 | 0,0% | 0,11 | 1,1% |
| Sample No.<br>8 | 11,9 | 0,07 | 0,6% | 0,08 | 0,6% | 0,00 | 0,0% | 0,06 | 0,5% | 0,16 | 1,4% | 0,00 | 0,0% | 0,20 | 1,7% |

(*) Total reproducibility includes : within-capillary, between-cay, between-day, between-ot and between-instrument.

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The reproducibility within the same instrument is summarized in the following tables:

- including within-capillary, between-capillary, between-run, between-day, between-lot and total reproducibility precision estimates (SD and %CV) for the HbA1c concentrations (in mmol/mol) and percentages for each instrument.
- including the within-lot precision estimates (SD and %CV) for the HbA10 concentrations -(in mmol/mol) and percentages for each lot on each instrument.

| Sample | Mean<br>(mmol/mol) | Within-capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|--------------------|------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,50 | 1,5% | 0,42 | 1,3% | 0,00 | 0,0% | 0,19 | 0,6% | 0,42 | 1,3% | 0,80 | 2,4% |
| Sample No. 2 | 34 | 0,42 | 1,3% | 0,49 | 1,4% | 0,00 | 0,0% | 0,28 | 0,8% | 0,62 | 1,8% | 0,94 | 2,8% |
| Sample No. 3 | 37 | 0,47 | 1,3% | 0,44 | 1,2% | 0,00 | 0,0% | 0,30 | 0,8% | 0,45 | 1,2% | 0,84 | 2,3% |
| Sample No. 4 | 48 | 0,55 | 1,1% | 0,38 | 0,8% | 0,18 | 0,4% | 0,28 | 0,6% | 0,49 | 1,0% | 0,89 | 1,9% |
| Sample No. 5 | 61 | 0,51 | 0,9% | 0,50 | 0,8% | 0,00 | 0,0% | 0,37 | 0,6% | 0,07 | 0,1% | 0,81 | 1,4% |
| Sample No. 6 | 65 | 0,59 | 0,9% | 0,38 | 0,6% | 0,00 | 0,0% | 0,34 | 0,5% | 0,38 | 0,6% | 0,86 | 1,3% |
| Sample No. 7 | 86 | 0,73 | 0,8% | 0,47 | 0,5% | 0,00 | 0,0% | 0,44 | 0,5% | 1,00 | 1,2% | 1,39 | 1,6% |
| Sample No. 8 | 107 | 0,70 | 0,6% | 0,76 | 0,7% | 0,00 | 0,0% | 0,70 | 0,6% | 2,41 | 2,2% | 2,71 | 2,5% |

#### Instrument No. 1

(*) Total reproducibility includes : within-capillary, between-run, between-run, between-day and between-lot.

| Sample | Mean (mmol/mol) | Within-lot (*) | | | | | |
|--------------|-----------------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,67 | 2,0% | 0,62 | 1,9% | 0,75 | 2,3% |
| Sample No. 2 | 34 | 0,72 | 2,1% | 0,75 | 2,3% | 0,65 | 1,9% |
| Sample No. 3 | 37 | 0,65 | 1,8% | 0,70 | 1,9% | 0,77 | 2,1% |
| Sample No. 4 | 48 | 0,80 | 1,7% | 0,78 | 1,6% | 0,70 | 1,5% |
| Sample No. 5 | 61 | 0,97 | 1,6% | 0,60 | 1,0% | 0,83 | 1,4% |
| Sample No. 6 | 65 | 0,76 | 1,2% | 0,64 | 1,0% | 0,93 | 1,4% |
| Sample No. 7 | 86 | 0,98 | 1,2% | 0,72 | 0,8% | 1,16 | 1,3% |
| Sample No. 8 | 107 | 1,33 | 1,3% | 1,20 | 1,1% | 1,25 | 1,2% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-day.

| Sample | Mean<br>(%) | Within-capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|-------------|------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,04 | 0,9% | 0,04 | 0,8% | 0,00 | 0,0% | 0,02 | 0,4% | 0,04 | 0,8% | 0,08 | 1,5% |
| Sample No. 2 | 5,2 | 0,04 | 0,8% | 0,05 | 0,9% | 0,00 | 0,0% | 0,03 | 0,6% | 0,05 | 0,9% | 0,08 | 1,6% |
| Sample No. 3 | 5,5 | 0,05 | 0,8% | 0,05 | 0,9% | 0,00 | 0,0% | 0,04 | 0,7% | 0,04 | 0,7% | 0,09 | 1,6% |
| Sample No. 4 | 6,5 | 0,05 | 0,7% | 0,04 | 0,6% | 0,02 | 0,3% | 0,02 | 0,2% | 0,05 | 0,7% | 0,08 | 1,2% |
| Sample No. 5 | 7,7 | 0,05 | 0,6% | 0,04 | 0.6% | 0,00 | 0,0% | 0,03 | 0,4% | 0,01 | 0.1% | 0,07 | 0,9% |
| Sample No. 6 | 8,1 | 0,06 | 0,7% | 0,04 | 0,5% | 0,00 | 0,0% | 0,03 | 0,4% | 0,03 | 0,4% | 0,08 | 1,0% |
| Sample No. 7 | 10,1 | 0,06 | 0,6% | 0,04 | 0,4% | 0,00 | 0,0% | 0,04 | 0,4% | 0,10 | 1,0% | 0,13 | 1,3% |
| Sample No. 8 | 11,9 | 0,07 | 0,6% | 0,07 | 0,6% | 0,00 | 0,0% | 0,06 | 0,5% | 0,21 | 1,8% | 0,24 | 2,0% |

510(k) Summary 7

{10}------------------------------------------------

(*) Total reproducibility includes : within-capillary, between-run, between-run, between-day and between-lot.

| Sample | Mean (%) | Within-lot (*) | | | | | |
|--------------|----------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,06 | 1,1% | 0,07 | 1,3% | 0,07 | 1,4% |
| Sample No. 2 | 5,2 | 0,06 | 1,2% | 0,08 | 1,6% | 0,06 | 1,2% |
| Sample No. 3 | 5,5 | 0,07 | 1,3% | 0,07 | 1,4% | 0,08 | 1,4% |
| Sample No. 4 | 6,5 | 0,07 | 1,1% | 0,06 | 0,9% | 0,07 | 1,0% |
| Sample No. 5 | 7,7 | 0,09 | 1,1% | 0,06 | 0,8% | 0,07 | 0,9% |
| Sample No. 6 | 8,1 | 0,08 | 0,9% | 0,06 | 0,7% | 0,09 | 1,1% |
| Sample No. 7 | 10,1 | 0,09 | 0,9% | 0,08 | 0,8% | 0,09 | 0,9% |
| Sample No. 8 | 11,9 | 0,12 | 1,0% | 0,11 | 0,9% | 0,12 | 1,0% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-day.

#### Instrument No. 2

| Sample | Mean<br>(mmol/mol) | Within-<br>capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|--------------------|----------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,57 | 1,7% | 0,55 | 1,7% | 0,00 | 0,0% | 0,22 | 0,7% | 0,45 | 1,4% | 0,94 | 2,8% |
| Sample No. 2 | 34 | 0,54 | 1,6% | 0,56 | 1,7% | 0,00 | 0,0% | 0,24 | 0,7% | 0,42 | 1,3% | 0,91 | 2,7% |
| Sample No. 3 | 37 | 0,45 | 1,2% | 0,53 | 1,5% | 0,00 | 0,0% | 0,23 | 0,6% | 0,31 | 0,8% | 0,80 | 2,2% |
| Sample No. 4 | 48 | 0,61 | 1,3% | 0,41 | 0,9% | 0,37 | 0,8% | 0,15 | 0,3% | 0,30 | 0,6% | 0,89 | 1,9% |
| Sample No. 5 | 61 | 0,55 | 0,9% | 0,28 | 0,5% | 0,00 | 0,0% | 0,48 | 0,8% | 0,00 | 0,0% | 0,78 | 1,3% |
| Sample No. 6 | 65 | 0,69 | 1,1% | 0,34 | 0,5% | 0,00 | 0,0% | 0,31 | 0,5% | 0,03 | 0,0% | 0,83 | 1,3% |
| Sample No. 7 | 86 | 0,58 | 0,7% | 0,55 | 0,6% | 0,00 | 0,0% | 0,40 | 0,5% | 0,41 | 0,5% | 0,99 | 1,1% |
| Sample No. 8 | 107 | 0,81 | 0,8% | 1,01 | 0,9% | 0,00 | 0,0% | 0,76 | 0,7% | 1,38 | 1,3% | 2,04 | 1,9% |

(*) Total reproducibility includes : within-capillary, between-run, between-day and between-lot.

| Sample | Mean<br>(mmol/mol) | Within-lot (*) | | | | | |
|--------------|--------------------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,88 | 2,6% | 0,73 | 2,2% | 0,84 | 2,5% |
| Sample No. 2 | 34 | 0,88 | 2,6% | 0,65 | 2,0% | 0,90 | 2,7% |
| Sample No. 3 | 37 | 0,87 | 2,4% | 0,56 | 1,5% | 0,74 | 2,0% |
| Sample No. 4 | 48 | 0,79 | 1,7% | 0,89 | 1,9% | 0,86 | 1,8% |
| Sample No. 5 | 61 | 0,66 | 1,1% | 0,67 | 1,1% | 0,98 | 1,6% |
| Sample No. 6 | 65 | 0,75 | 1,2% | 0,77 | 1,2% | 0,99 | 1,5% |
| Sample No. 7 | 86 | 1,03 | 1,2% | 0,79 | 0,9% | 0,86 | 1,0% |
| Sample No. 8 | 107 | 1,73 | 1,6% | 1,45 | 1,4% | 1,47 | 1,4% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-day.

{11}------------------------------------------------

| Sample | Mean<br>(%) | Within-capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|-------------|------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,05 | 1,0% | 0,05 | 0,9% | 0,00 | 0,0% | 0,02 | 0,4% | 0,05 | 0,9% | 0,09 | 1,7% |
| Sample No. 2 | 5,2 | 0,05 | 0,9% | 0,06 | 1,1% | 0,00 | 0,0% | 0,02 | 0,4% | 0,03 | 0,7% | 0,09 | 1,6% |
| Sample No. 3 | 5,5 | 0,04 | 0,8% | 0,04 | 0,8% | 0,00 | 0,0% | 0,02 | 0,4% | 0,04 | 0,8% | 0,08 | 1,4% |
| Sample No. 4 | 6,5 | 0,05 | 0,7% | 0,03 | 0,5% | 0,02 | 0,3% | 0,02 | 0,4% | 0,02 | 0,4% | 0,07 | 1,1% |
| Sample No. 5 | 7,7 | 0,05 | 0,7% | 0,03 | 0,3% | 0,00 | 0,0% | 0,05 | 0,6% | 0,01 | 0,1% | 0,08 | 1,0% |
| Sample No. 6 | 8,1 | 0,07 | 0,8% | 0,03 | 0,3% | 0,01 | 0,1% | 0,03 | 0,3% | 0,01 | 0,1% | 0,08 | 0,9% |
| Sample No. 7 | 10,1 | 0,05 | 0,5% | 0,05 | 0,5% | 0,00 | 0,0% | 0,03 | 0,3% | 0,05 | 0,5% | 0,10 | 1,0% |
| Sample No. 8 | 11,9 | 0,08 | 0,6% | 0,09 | 0,8% | 0,02 | 0,2% | 0,07 | 0,6% | 0,13 | 1,1% | 0,19 | 1,6% |

(*) Total reproducibility includes : within-capillary, between-run, between-day and between-lot.

| Sample | Mean (%) | Within-lot (*) | | | | | |
|--------------|----------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,08 | 1,5% | 0,06 | 1,2% | 0,08 | 1,6% |
| Sample No. 2 | 5,2 | 0,09 | 1,7% | 0,07 | 1,3% | 0,08 | 1,6% |
| Sample No. 3 | 5,5 | 0,06 | 1,2% | 0,06 | 1,1% | 0,07 | 1,3% |
| Sample No. 4 | 6,5 | 0,06 | 1,0% | 0,06 | 1,0% | 0,07 | 1,1% |
| Sample No. 5 | 7,7 | 0,06 | 0,8% | 0,06 | 0,8% | 0,10 | 1,2% |
| Sample No. 6 | 8,1 | 0,07 | 0,8% | 0,08 | 0,9% | 0,09 | 1,1% |
| Sample No. 7 | 10,1 | 0,10 | 1,0% | 0,07 | 0,7% | 0,08 | 0,8% |
| Sample No. 8 | 11,9 | 0,15 | 1,3% | 0,14 | 1,2% | 0,14 | 1,2% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-run and between-day.

#### Instrument No. 3

| Sample | Mean<br>(mmol/mol) | Within-capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|--------------------|------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,44 | 1,3% | 0,46 | 1,4% | 0,00 | 0,0% | 0,30 | 0,9% | 0,33 | 1,0% | 0,77 | 2,3% |
| Sample No. 2 | 34 | 0,49 | 1,4% | 0,50 | 1,5% | 0,00 | 0,0% | 0,32 | 0,9% | 0,40 | 1.2% | 0,87 | 2,6% |
| Sample No. 3 | 37 | 0,40 | 1,1% | 0,54 | 1,5% | 0,00 | 0,0% | 0,29 | 0,8% | 0,48 | 1,3% | 0,88 | 2,4% |
| Sample No. 4 | 48 | 0,52 | 1,1% | 0,49 | 1,0% | 0,17 | 0,4% | 0,21 | 0,4% | 0,32 | 0,7% | 0,83 | 1,7% |
| Sample No. 5 | 61 | 0,54 | 0,9% | 0,46 | 0,7% | 0,00 | 0,0% | 0,42 | 0,7% | 0,15 | 0,2% | 0,84 | 1,3% |
| Sample No. 6 | 65 | 0,59 | 0,9% | 0,55 | 0,8% | 0,00 | 0,0% | 0,28 | 0,4% | 0,12 | 0,2% | 0,86 | 1,3% |
| Sample No. 7 | 86 | 0,58 | 0,7% | 0,45 | 0,5% | 0,00 | 0,0% | 0,31 | 0,4% | 0,61 | 0,7% | 1,01 | 1,2% |
| Sample No. 8 | 107 | 0.69 | 0,6% | 0,68 | 0,6% | 0,00 | 0,0% | 0,53 | 0,5% | 1,39 | 1,3% | 1,77 | 1,7% |

(*) Total reproducibility includes : within-capillary, between-run, between-day and between-lot.

{12}------------------------------------------------

| Sample | Mean (mmol/mol) | Within-lot (*) | | | | | |
|--------------|-----------------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 33 | 0,61 | 1,8% | 0,89 | 2,7% | 0,57 | 1,7% |
| Sample No. 2 | 34 | 0,64 | 1,9% | 1,00 | 3,0% | 0,59 | 1,7% |
| Sample No. 3 | 37 | 0,53 | 1,4% | 0,94 | 2,6% | 0,67 | 1,8% |
| Sample No. 4 | 48 | 0,72 | 1,5% | 0,88 | 1,8% | 0,74 | 1,5% |
| Sample No. 5 | 61 | 0,78 | 1,2% | 0,87 | 1,4% | 0,83 | 1,3% |
| Sample No. 6 | 65 | 0,71 | 1,1% | 0,88 | 1,4% | 0,96 | 1,5% |
| Sample No. 7 | 86 | 0,69 | 0,8% | 0,76 | 0,9% | 0,93 | 1,1% |
| Sample No. 8 | 107 | 1,09 | 1,0% | 1,16 | 1,1% | 1,09 | 1,0% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-day.

| Sample | Mean<br>(%) | Within-capillary | | Betweencapillary | | Between-run | | Between-day | | Between-lot | | Total<br>reproducibility<br>(*) | |
|--------------|-------------|------------------|------|------------------|------|-------------|------|-------------|------|-------------|------|---------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,05 | 0,9% | 0,04 | 0,8% | 0,00 | 0,0% | 0,03 | 0,5% | 0,01 | 0,2% | 0,07 | 1,3% |
| Sample No. 2 | 5,2 | 0,05 | 0,9% | 0,04 | 0,8% | 0,00 | 0,0% | 0,03 | 0,6% | 0,03 | 0,6% | 0,08 | 1,5% |
| Sample No. 3 | 5,5 | 0,04 | 0,8% | 0,04 | 0,7% | 0,00 | 0,0% | 0,02 | 0,4% | 0,04 | 0,7% | 0,07 | 1,3% |
| Sample No. 4 | 6,5 | 0,05 | 0,8% | 0,04 | 0,6% | 0,02 | 0,4% | 0,02 | 0,2% | 0,03 | 0,4% | 0,07 | 1,1% |
| Sample No. 5 | 7,7 | 0,05 | 0,6% | 0,04 | 0,6% | 0,00 | 0,0% | 0,03 | 0,4% | 0,02 | 0,3% | 0,08 | 1,0% |
| Sample No. 6 | 8,1 | 0,06 | 0,7% | 0,05 | 0,6% | 0,00 | 0,0% | 0,03 | 0,4% | 0,01 | 0,1% | 0,08 | 1,0% |
| Sample No. 7 | 10,1 | 0,05 | 0,5% | 0,04 | 0,4% | 0,00 | 0,0% | 0,03 | 0,3% | 0,06 | 0,6% | 0,10 | 1,0% |
| Sample No. 8 | 11,9 | 0,06 | 0,5% | 0,07 | 0,6% | 0,00 | 0,0% | 0,05 | 0,4% | 0,13 | 1,1% | 0,17 | 1,4% |

(*) Total reproducibility includes : within-capillary, between-run, between-run, between-lot.

| Sample | Mean<br>(%) | Within-lot (*) | | | | | |
|--------------|-------------|----------------|------|-----------|------|-----------|------|
| | | Lot No. 1 | | Lot No. 2 | | Lot No. 3 | |
| | | SD | CV | SD | CV | SD | CV |
| Sample No. 1 | 5,2 | 0,05 | 1,0% | 0,09 | 1,6% | 0,06 | 1,2% |
| Sample No. 2 | 5,2 | 0,06 | 1,2% | 0,09 | 1,7% | 0,06 | 1,2% |
| Sample No. 3 | 5,5 | 0,05 | 0,9% | 0,08 | 1,4% | 0,06 | 1,1% |
| Sample No. 4 | 6,5 | 0,07 | 1,0% | 0,08 | 1,2% | 0,06 | 1,0% |
| Sample No. 5 | 7,7 | 0,07 | 0,9% | 0,08 | 1,0% | 0,07 | 0,9% |
| Sample No. 6 | 8,1 | 0,07 | 0,9% | 0,08 | 1,0% | 0,09 | 1,2% |
| Sample No. 7 | 10,1 | 0,06 | 0,6% | 0,07 | 0,7% | 0,10 | 0,9% |
| Sample No. 8 | 11,9 | 0,11 | 0,9% | 0,11 | 0,9% | 0,10 | 0,9% |

(*) Within-lot reproducibility includes : within-capillary, between-run and between-day.

{13}------------------------------------------------

### b. Linearity / assay reportable range

A linearity study was performed per CLSI EP06-A: Evaluation of Quantitative Measuring Procedures: A Statistical Approach, Linearity across the reportable range was performed using low 3.8% Hb A1c (18 mmol/mol) and high 17.3% (166 mmol/mol).

### Mixture of 2 different blood samples:

2 characteristic blood samples, including a normal sample and an elevated HbA10 level sample were mixed within different proportions and the mixtures were electrophoresed with the CAPI 3 Hb A1c procedure. For each mixture, samples were analyzed in triplicate.

The tests were determined to be linear within the entire range studied for HbA% hemoglobin fraction. The stated measuring range is 20 mmol/mol to 157 mmol/mol HbA+c (4.0 % to 16.5 % HbA1c).

### Dilution of 4 different blood samples in hemolysing solution :

4 different characteristic blood samples, including 1 normal sample with HbA1c concentration at 21 mmol/mol (4.1 % HbA1c), 1 sample with HbA1c level close to the cut-off value with HbA1c concentration at 47 mmol/mol (6.4 % HbA1c) and 2 elevated HbA1c level samples with HbA1c concentrations at 82 mmol/mol (9.6 % HbA1c) and at 134 mmol/mol (14.4 % HbA1c), were all serially diluted in hemolysing solution and electrophoresed with the CAPI 3 Hb A1c procedure. The tests were determined to be linear within the entire ranges studied from 2.9 to 30.5 g/dL total hemoglobin and HbA1c fraction concentration and percentage were not affected by the hemoglobin concentration of the samples.

### c. Traceability, Stability (controls, calibrators, or methods)

The CAPI 3 Hb A1c test standardization is traceable to the International Federation of Clinical Chemistry (IFCC) reference calibrators.

The CAPI 3 Hb A1c assay is NGSP certified. The NGSP certification expires in one year. See the NGSP website for current certification at http://www.ngsp.org

Hb A1c results are provided in two different units: NGSP equivalent units (%) and IFCC equivalent units (mmol/mol).

### d. Calibrators and Controls

The Hb A1c CAPILLARY CALIBRATORS are required for use with this device. Value assignment and stability protocal and acceptance criteria were previously reviewed and cleared in k162281 and k122101.

Sebia MULTI-SYSTEM Hb A1c CAPILLARYS CONTROLS are required for use with this device and cleared in submission k162281.

{14}------------------------------------------------

## e. Comparison Studies

A method comparison study of 152 variant-free whole blood samples covering the measuring range were evaluated using the Capi3 Hb A1c kit and the CAPILLARYS 3 TERA instrument. The results were compared to the testing performed at a NGSP reference laboratory using the cleared HPLC HbA1c method (Automated Glycohemoglobin Analyzer HLC-712G8).

To support the diagnostic claim for HbA1c the samples spanned around the decision points as follows:

| HbA1c level | Number of samples | % of samples |
|-----------------------|-------------------|--------------|
| HbA1c ≤ 5 % | 9 | 6 |
| 5,0 % < HbA1c ≤ 6,0 % | 19 | 13 |
| 6,0 % < HbA1c ≤ 6,5 % | 35 | 23 |
| 6,5 % < HbA1c ≤ 7,0 % | 36 | 24 |
| 7,0 % < HbA1c ≤ 8,0 % | 23 | 15 |
| 8,0 %< HbA1c ≤ 9,0 % | 13 | 9 |
| HbA1c > 9,0 % | 17 | 11 |
| Total | 152 | 100 |

HbA1c results given as a NGSP unit (%)

| Fraction | Correlation coefficient | y-intercept | Slope | Range of HbA1c % values<br>CAPI 3 Hb A1c |
|----------|-------------------------|-------------|-------|------------------------------------------|
| HbA1c | 0,999 | -0,142 | 1,014 | 3,9 - 16,5 |

152 samples

| Regression Analysis (HbA1c fraction) | | | | |
|----------------------------------------------------|-------------------------------------------|--|--|--|
| Ordinary line ar fit | | | | |
| 95 % Confidence Intervals are shown in parentheses | | | | |
| Points (Plotted/Total) | 152/152 | | | |
| Results Ranges | 3,9 to 16,5 | | | |
| Normal range | < 6,5 | | | |
| Correlation coefficient (r) | 0,999 | | | |
| Slope | 1,014 (1,007 to 1,021) | | | |
| y-intercept | -0,142 (-0,197 to -0,087) | | | |
| Average bias (all samples) | -0,04 (-0,06 to -0,02) | | | |
| Bias at normal range | The bias at 6,5 is -0,05 (-0,07 to -0,03) | | | |

The unit of the bias is the same of the result provided (%)

{15}------------------------------------------------

### f. Total Error Calculations

Total error (TE) is calculated for 4 concentrations, corresponding to the concentrations of the samples used in the reproducibility study, (5.2 %, 6.5 %, 8.1 % and 11.9 %) using the results of bias estimation (%Bias) and coefficients of variation (CV).

Total Error is calculated as follows: %TE=|%Bias| + 1.96 *CV* (1 + %Bias/100)).

The results are presented in the following table for the CAPI3 Hb A1c using the CAPILLARYS 3 TERA instrument:

| Linear equation<br>regression | Target (%) | y from linear<br>equation | Deviation from<br>target | %Bias | CV (%) | TE (%) | Specification |
|-------------------------------|------------|---------------------------|--------------------------|-------|--------|--------|---------------|
| $y = 1,014 x -0,142$ | 5,2 | 5,1 | -0,1 | -1,32 | 1,6 | 4,4 | ≤ 6,0 % |
| | 6,5 | 6,4 | -0,1 | -0,77 | 1,1 | 2,9 | |
| | 8,1 | 8,1 | 0,0 | -0,34 | 1,0 | 2,3 | |
| | 11,9 | 11,9 | 0,0 | 0,22 | 1,7 | 3,6 | |

HbA1c results given as a NGSP unit (%)

### g. Interferences

Interference studies were performed per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Each study was performed using 2 different whole blood samples: a blood sample close to the cut-off value and a blood sample with elevated HbA1c level. Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system.

No interference with the CAPI 3 Hb A1c procedure was detected due to the blood sample's high concentration of the following interfering factors tested at levels equal to the concentrations listed below:

| Endogenous interfering factor | Concentration |
|-------------------------------|---------------------|
| Bilirubin | 70 mg/dL (1197 µM) |
| D-glucose | 1000 mg/dL (55 mM) |
| Glycated Albumin | 2.2 mg/mL |
| Rheumatoid factor | 1294 IU/mL |
| Total Protein | 149.5 g/L |
| Triglycerides | 3.58 g/dL (40.9 mM) |
| Urea | 265 mg/dL (44.2 mM) |

| Drug | Concentration |
|----------------------|-----------------------|
| Acetaminophen | 200 mg/L (1325 μM) |
| Acetylcysteine | 200 mg/dL (12.3 mM) |
| Acetylsalicilyc acid | 1000 mg/dL (55.56 mM) |
| Ampicillin-Na | 1000 mg/dL (28653 μM) |
| Ascorbic acid | 300 mg/dL (17045 μM) |
| Cefoxitin | 2500 mg/dL (58548 μM) |
| Cyclosporine | 5 mg/L |

510(k) Summary 13

{16}------------------------------------------------

| Doxycycline | 50 mg/dL (1123.6 μM) |
|----------------|----------------------|
| Glybenclamide | 3 mg/dL |
| Heparin | 5000 U/L |
| Ibuprofen | 500 mg/L (2427 μM) |
| Levodopa | 40 mg/dL |
| Metformin | 5 mg/dL |
| Methyldopa | 40 mg/dL (1896 μM) |
| Metronidazole | 200 mg/dL (11696 μM) |
| Phenylbutazone | 400 mg/L |
| Rifampicin | 70 mg/L (85.1 μM) |
| Theophylline | 100 mg/L (556 μM) |

| Hemoglobin Derivatives and cross<br>reactants | Concentrations |
|-----------------------------------------------|----------------|
| Carbamylated Hemoglobin | ≤ 8.1 mg/mL |
| HbA1a+b | ≤ 0.20 mg/mL |
| Labile HbA1c | ≤ 20.0 mg/mL |
| Acetylated hemoglobin | ≤ 4.2 mg/mL |
| Glycated Albumin | ≤ 2.2 mg/mL |

Hemoglobin Variant Study was performed using specific samples known to contain hemoglobin variants S, C, E, D, A2 and F. The samples were analyzed with a reference method performed in a NGSP laboratory (reference) and with CAPI 3 Hb A1c procedure on CAPILLARYS 3 TERA instrument (test): percentages of HbA1c fraction. The relative deviation (%) between the reference procedure and the test procedure has been calculated for each sample (see the following tables).

| Hemoglobin<br>Variants | No.of<br>Samples | Ranges in %<br>Variant | Range of % HbA1c<br>Concentration |
|------------------------|------------------|------------------------|-----------------------------------|
| Hb A2 | 20 | 4.0-7.8 | 4.5-11.2 |
| Hb F | 19 | 1.5-23.7 | 5.1-15.1 |
| Hb S | 24 | 33.0-40.8 | 4.9-14.7 |
| Hb C | 24 | 25.6-37.6 | 4.9-12.4 |
| Hb D | 24 | 35.5-41.3 | 5.2-12.0 |
| Hb E | 20 | 21.1-26.8 | 4.9-9.6 |

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The percent relative bias from a reference method (commercially available high-performance liquid chromatography technique for HbAre quantification) at low and high concentrations of HbA1c in each sample is:

| Hemoglobin<br>Variant | Relative % Bias from<br>Reference Method ~ 6.5 % Hb A1c<br>Mean Relative % Bias<br>(Range of % Bias for Hb A1c) | Relative % Bias from<br>Reference Method ~ 9 % Hb A1c<br>Mean Relative % Bias<br>(Range of % Bias for Hb A1c) |
|-----------------------|-----------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------|
| Hb S | 1.6% (0% - 3.2%) | 2.9% (1.1% - 5.4%) |
| Hb C | -1.8% (-3.1% - 0%) | 3.9% (3.3% - 4.5%) |
| Hb D | 1.0% (0% - 1.6%) | 0.8% (0% - 2.4%) |
| Hb E | 1.5% (-1.5% - 1.6%) | 1.2% (0%-2.5%) |
| Hb A2 | 0.7% (0%-1.5%) | 0.4% (0%-1.1%) |
| Hb F | -4.1% (-3.3% -4.9 %) | -0.8% (0%-2.0%) |

- A significant neqative interference has been observed with fetal hemoglobin (HbF) concentrations > 23%. HbA1c results are invalid for patients with high amounts of HbF (>23%) including those with known Hereditary Persistence of Fetal Hemoglobin

- Levels of Hb A2 up to 7.8 % in the blood sample do not interfere with HbA1c fraction quantification.

- No interference has been observed with HbA1c fraction quantification due to the presence of major abnormal hemoglobins Hb S (≤ 40.8 %), Hb C (≤ 37.6 %), Hb D (≤ 41.3 %) and Hb E (≤ 26.8 %).

## h. Expected Values/ Reference Range

Hemoglobin Are expected value range was cited from the American Diabetes Association (Standards of Medical Care in Diabetes 2016, 39 (Suppl 1)). The American Diabetes Association's (ADA) most recent Clinical Practice are:

| Category | HbA1c Range (IFCC) | HbA1c Range (NGSP/DCCT) |
|----------------------------------------------|---------------------------|-------------------------|
| Normal | < 39 mmol/mol | < 5.7 % |
| Prediabetes (increased risk for<br>diabetes) | 39 mmol/mol - 47 mmol/mol | 5.7 % - 6.4 % |
| Diabetes | ≥ 48 mmol/mol | ≥ 6.5 % |

The expected HbA ; range for non-diabetic adults is 20 = 42 mmol/mol or 4.0 - 6.0 %. However, each laboratory should establish its reference range and HbA1c goal in their country of business taking into account sex, age, ethnicity and individual patient situation.

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## 6. Special Controls for Diabetes Diagnosis Claim

| YES OR NO | Special Controls |
|-----------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| YES | Device must have initial and annual standardization verification by<br>certifying glycohemoglobin standardization organization deemed<br>acceptable by FDA |
| YES | Performance testing of device precision must, at a minimum use blood<br>samples with concentrations near 5.0%, 6.5%, 8.0% and 12 %<br>hemoglobin A1c. Testing must evaluate precision over a minimum of<br>20 days using at least 3 lots of the device and instruments, as<br>applicable |
| YES | Performance testing of accuracy must include a minimum of 120<br>blood samples that span the measuring interval of the new device<br>and compare results of the new device to results of the standardized<br>method. Results must demonstrate little or no bias versus the<br>standardized method. |
| YES | Total error of the new device must be evaluated using single<br>measurements by the new device compared to the results of the<br>standardized test method, and this evaluation must demonstrate a<br>total error of less than or equal to 6% |
| YES | Performance testing must demonstrate that there is little to no<br>interference from common hemoglobin variants, including<br>Hemoglobin C, Hemoglobin D, Hemoglobin E, Hemoglobin A2 and<br>Hemoglobin S. |
| NA | When assay interference from Hemoglobin F or interference with<br>other hemoglobin variants with low frequency in the population is<br>observed, a warning statement must be placed in a black box and<br>must appear in all the labeling material for these devices describing<br>the interference and any affected population. |

## 7. Conclusion:

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