D-10 Hemoglobin A1c Program

K161687 · Bio-Rad Laboratories, Inc. · PDJ · Oct 14, 2016 · Clinical Chemistry

Device Facts

• Record ID: K161687
• Device Name: D-10 Hemoglobin A1c Program
• Applicant: Bio-Rad Laboratories, Inc.
• Product Code: PDJ · Clinical Chemistry
• Decision Date: Oct 14, 2016
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1373
• Device Class: Class 2

Intended Use

The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/ mol and NGSP %) in human whole blood using ion- exchange high-performance liquid chromatography (HPLC) on the D-10TM Hemoglobin Testing System. Hemoglobin Alc measurements are 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 D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.

Device Story

The D-10 Hemoglobin A1c Program is an in vitro diagnostic assay for professional use in clinical settings. It utilizes ion-exchange high-performance liquid chromatography (HPLC) to analyze human whole blood samples. The system uses a dual-piston pump to deliver buffer gradients through an analytical cartridge, separating hemoglobin species based on ionic interactions. Absorbance is measured at 415 nm via a photometer flow cell, producing a digital chromatogram. Software processes raw data, potentially applying calibration factors, to generate sample reports. The device aids clinicians in diagnosing diabetes, identifying at-risk patients, and monitoring long-term glycemic control. It provides quantitative HbA1c results in both NGSP (%) and IFCC (mmol/mol) units, enabling informed clinical decision-making regarding patient diabetes management.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated per CLSI EP05-A2 (60-day study, 720 results/sample). Linearity demonstrated from 3.9–18.8% HbA1c (NGSP) and 19–182 mmol/mol (IFCC). Method comparison (n=128) against NGSP SRL reference method showed strong correlation (Deming slope 0.9722, R²=1.0000). Interference studies confirmed no significant impact from endogenous substances (bilirubin, glucose, lipids, protein, rheumatoid factor), common drugs, or hemoglobin variants (S, C, D, E, A2) at tested levels. Significant positive interference noted for HbF >10%.

Technological Characteristics

Ion-exchange HPLC; dual-piston low-pulsation pump; cation exchange analytical cartridge (4.0 mm ID x 30 mm); photometer flow cell (415 nm detection). Reagents include elution buffers (Bis-Tris/Phosphate) and calibrators (lyophilized human RBC hemolysate). Connectivity via D-10 Hemoglobin Testing System. Software performs data reduction and chromatogram generation. Preservatives: <0.05% sodium azide.

Indications for Use

Indicated for professional in vitro diagnostic use in adults for the diagnosis of diabetes, identification of patients at risk for diabetes, and monitoring long-term blood glucose control in patients with diabetes mellitus. Not for use with newborns. Not for use to replace glucose testing in pediatric patients, pregnant women, or patients with Type 1 diabetes. Not for use to diagnose diabetes during pregnancy or gestational diabetes. Not for use in patients with conditions altering red blood cell lifespan (e.g., hemolytic anemia, hemoglobinopathies, thalassemias, recent blood loss/transfusion, iron deficiency) or severe hepatic/renal disease.

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

• VARIANT II TURBO HbA1c Kit -2.0 (k142448)

Submission Summary (Full Text)

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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 is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter of the circle. In the center of the circle is a stylized image of three human profiles facing to the right.

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

BIO-RAD LABORATORIES, INC. JACKIE BUCKLEY REGULATORY AFFAIRS SUPERVISOR 4000 ALFRED NOBEL DR. HERCULES CA 94547

October 14, 2016

Re: K161687

Trade/Device Name: D-10 Hemoglobin A1c Program Regulation Number: 21 CFR 862.1373 Regulation Name: Glycosylated hemoglobin assay Regulatory Class: II Product Code: PDJ Dated: August 30, 2016 Received: September 1, 2016

Dear Ms. Buckley:

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.

# Katherine Serrano -S

For: 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) k161687

Device Name D-10TM Hemoglobin A1c Program

#### Indications for Use (Describe)

The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/ mol and NGSP %) in human whole blood using ion- exchange high-performance liquid chromatography (HPLC) on the D-10TM Hemoglobin Testing System.

Hemoglobin Alc measurements are 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 D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.

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

| <span></span> | <span></span> |
|----------------------------------------------|---------------|
| <div></div> | <div></div> |
| Prescription Use (Part 21 CFR 801 Subpart D) | <div></div> |
| Over-The-Counter Use (21 CFR 801 Subpart C) | <div></div> |

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## 510(k) Summary (Summary of Safety and Effectiveness)

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

The assigned 510(k) number is: K161687

Date Summary prepared: Oct. 12, 2016

#### 1. Applicant Name:

Bio-Rad Laboratories, Inc. Clinical Diagnostics Group 4000 Alfred Nobel Drive Hercules, California 94547

#### 2. Contact Person(s):

Jackie Buckley, Regulatory Affairs Supervisor Telephone Number: [phone] FAX: [phone] E-Mail: jackie [e-mail]

Alfred Evans, RA/QA Director Telephone Number: [phone] FAX: [phone] E-Mail:al [e-mail]

#### 3. Device Name/Trade Name:

Reagents: Trade Name: D-10™ Hemoglobin A1c Program Classification Name: Hemoglobin A1c Test System Common Name: HbA1c Product Code: PDJ C.F.R Section: 21 CFR 862.1373 Device classification: Class II Panel Classification: Chemistry

#### 4. Predicate Device:

| Predicate Device<br>Name | Predicate Device<br>510(k) Number |
|---------------------------------|-----------------------------------|
| VARIANT II TURBO HbA1c Kit -2.0 | K142448 |

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#### 5. Description of the Device:

The D-10™ Hemoglobin Testing System utilizes the principles of ion-exchange highperformance liquid chromatography (HPLC). A dual-piston, low pulsation HPLC pump and a proportioning value deliver the buffer solution to an analytical cartridge and detector. Whole blood samples undergo an automatic two step dilution process and then introduced into the analytical flow path. Pre-diluted samples are aspirated directly and introduced into the analytical flow path. Between sample injections, the sample probe is rinsed with Wash/Diluent Solution to minimize sample carryover.

A programmed buffer gradient of increasing ionic strength delivers the sample to the analytical cartridge where the hemoglobin species are separated based upon their ionic interactions with the cartridge material and the buffer gradient. The separated hemoglobin species then pass through the photometer flow cell where changes in the absorbance are measured at 415 nm and recorded as a digital chromatogram.

The software performs a reduction of raw data collected from each analysis that may indicate use of a calibration factor. A samples report and chromatogram are generated for each sample.

The D-10™ Hemoglobin A1c Program is designed to be used on the D-10™ Hemoglobin Testing System with or without a D-10 Rack Loader.

#### Reagents:

The D-10™ HbA1c reagents contain the following components: Description

D-10 ™ Hemoglobin Are Analytical Cartridge. Cation exchange cartridge (400 tests), 4.0 mm ID x 30 mm.

D-10 ™ Wash/Diluent Solution. Each bottle contains 1600 mL of deionized water with <0.05 sodium azide as a preservative.

Floppy diskette with D-10 Hemoglobin A1c "Update Kit" program parameters.

D-10™ Hemoglobin Are Elution Buffer 1. Each bottle contains 2000 mL of a Bis-Tris/Phosphate buffer. Contains <0.05% sodium azide as a preservative.

D-10 "" Hemoglobin Are Elution Buffer 2. Each bottle contains 1000 mL of a Bis-Tris/Phosphate buffer. Contains<0.05% sodium azide as a preservative.

D-10 10 10 Hemoglobin A16 Calibrator/Diluent Set. One set consisting of 3 vials of Calibrator 1, 3 vials of Calibrator 2, and 1 bottle of Calibrator Diluent. The calibrator vials contain Ivophilized human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservative. Reconstituted volume is 7mL per vial.

Calibrator Diluent contains 100 mL of deionized water with <0.05% sodium azide as preservative.

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CD-ROM with D-10 Hemoglobin A1c "Update Kit" program parameters

Whole Blood Primer. Each vial contains lyophilized human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservatives. Reconstituted volume is 1.0 mL per vial.

Sample Vials. 100 polypropylene microvials with pierceable caps, 1.5mL

D-10 Hemoglobin A1c Supplemental Reagent Pack. This reagent pack is used as a supplement to the D-10 Hemoglobin A1c Reorder Pack.

#### 6. Indications for Use:

The D-10™ Hemoglobin A1c Program is intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) in human whole blood using ionexchange high-performance liquid chromatography (HPLC) on the D-10™ Hemoglobin Testing System.

Hemoqlobin A1c measurements are 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 D-10™ Hemoglobin A1c Program is intended for professional in vitro diagnostic use only.

#### 7. Substantial Equivalence Information:

#### Predicate Device Information:

| Predicate Device | Predicate Device |
|---------------------------------|------------------|
| Name | 510(k) Number |
| VARIANT II TURBO HbA1c Kit -2.0 | K142448 |

The comparison of the technological characterizes of the D-10 Hemoglobin A1c Program (candidate assay) utilizes principles of ion-exchange high-performance liquid chromatography (HPLC) similar to the same technology of the VARIANT II TURBO HbA1c Kit - 2.0 (predicate device).

Tables 1 and 2 provide the similarities and differences between the candidate assay and the predicate assay.

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| Reagent<br>Similarities and Differences | | |
|-----------------------------------------|----------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Features | Candidate Device:<br>D-10™ Hemoglobin A1c<br>(K)161687 | Predicate Device:<br>VARIANT™ II TURBO HbA1c<br>Kit - 2.0<br>(K)142448 |
| Intended Use | Same | Intended for the<br>quantitative determination of<br>hemoglobin A1c (IFCC mmol/mol<br>and NGSP %) |
| Platform | D-10™ Hemoglobin Testing<br>System | VARIANT™II TURBO<br>Hemoglobin Testing System and<br>VARIANT™II TURBO Link<br>Hemoglobin Testing System |
| Measuring<br>Interval | 3.9 to 18.8% (NSGP)<br>19 – 182 mmol/mol<br>HbA1c (IFCC) | 3.4 to 20.6 % (NSGP)<br>14 – 203 mmol/mol<br>HbA1c (IFCC) |
| Specimen Type | Same | Human Whole blood |
| Assay Principle | Same | Ion exchange HPLC |
| Matrices | K₂-EDTA, K₃-EDTA | K₂-EDTA, K₃-EDTA<br>Hemoglobin Capillary Collection Kit |
| Standardization | Same | Traceable to the Diabetes<br>Control and Complications Trial<br>(DCCT) reference method and<br>IFCC. Certified via the National<br>Glycohemoglobin Standardization<br>Program (NGSP) |

## Table 1: Reagent Similarities and Differences

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## 8. Summary of Nonclinical Performance Data:

#### Precision/Reproducibility: a.

The precision of the D-10™ Hemoglobin A1c Program was evaluated based on CLSI EP05-A2 guidelines, Evaluation of Precision Performance of Quantitative Measurement Methods using a modified study design. Four EDTA whole blood samples at the following targeted HbA1c concentrations of ~5%, ~6.5%, ~8% and ~12% were utilized in the study. In addition, five quality control materials were also tested. The samples were run in duplicate in 2 runs per day on 3 instruments for 20 days. The study was repeated using 3 different kit lots, yielding a total of 720 results per sample over a 60-day period. NGSP results are shown in Tables 2-5. IFCC results are shown in Tables 6-9.

| | HbA1c,% | | Between-Run | | Between-Day | | Between-Lot | | Total Precision | | |
|-----------|---------|---------------|-------------|------|-------------|------|-------------|------|-----------------|------|-----|
| Sample ID | | Repeatability | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 5.2 | 0.07 | 1.3 | 0.00 | 0.0 | 0.04 | 0.7 | 0.08 | 1.6 | 0.11 | 2.2 |
| Patient 2 | 6.7 | 0.07 | 1.0 | 0.00 | 0.0 | 0.07 | 1.1 | 0.05 | 0.7 | 0.11 | 1.7 |
| Patient 3 | 8.4 | 0.05 | 0.6 | 0.01 | 0.2 | 0.08 | 1.0 | 0.02 | 0.2 | 0.10 | 1.2 |
| Patient 4 | 12.7 | 0.09 | 0.7 | 0.06 | 0.5 | 0.17 | 1.3 | 0.21 | 1.7 | 0.29 | 2.3 |
| Control 1 | 5.6 | 0.07 | 1.2 | 0.00 | 0.0 | 0.05 | 0.9 | 0.05 | 0.9 | 0.10 | 1.8 |
| Control 2 | 10.4 | 0.10 | 0.9 | 0.00 | 0.0 | 0.12 | 1.1 | 0.12 | 1.2 | 0.20 | 1.9 |
| QC 1 | 5.5 | 0.06 | 1.0 | 0.00 | 0.0 | 0.05 | 0.9 | 0.08 | 1.5 | 0.11 | 2.1 |
| QC 2 | 10.0 | 0.13 | 1.3 | 0.00 | 0.0 | 0.09 | 0.9 | 0.06 | 0.6 | 0.17 | 1.7 |
| QC 3 | 15.6 | 0.11 | 0.7 | 0.15 | 1.0 | 0.08 | 0.5 | 0.17 | 1.1 | 0.27 | 1.7 |

## Table 2: Instrument 1 % CV by Sample (NGSP)

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| Sample ID | HbA1c,<br>% | Repeatability | | Between-<br>Run | | Between-<br>Day | | Between-Lot | | Total<br>Precision | |
|-----------|-------------|---------------|-----|-----------------|-----|-----------------|-----|-------------|-----|--------------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 5.2 | 0.04 | 0.9 | 0.02 | 0.4 | 0.04 | 0.8 | 0.08 | 1.5 | 0.10 | 1.9 |
| Patient 2 | 6.6 | 0.03 | 0.5 | 0.04 | 0.6 | 0.05 | 0.8 | 0.02 | 0.3 | 0.08 | 1.2 |
| Patient 3 | 8.2 | 0.04 | 0.5 | 0.04 | 0.4 | 0.07 | 0.8 | 0.02 | 0.2 | 0.09 | 1.0 |
| Patient 4 | 12.5 | 0.06 | 0.5 | 0.08 | 0.6 | 0.13 | 1.1 | 0.21 | 1.7 | 0.27 | 2.1 |
| Control 1 | 5.6 | 0.04 | 0.8 | 0.02 | 0.3 | 0.06 | 1.0 | 0.07 | 1.2 | 0.10 | 1.7 |
| Control 2 | 10.1 | 0.04 | 0.4 | 0.06 | 0.6 | 0.08 | 0.8 | 0.09 | 0.9 | 0.14 | 1.4 |
| QC 1 | 5.5 | 0.03 | 0.6 | 0.02 | 0.4 | 0.06 | 1.0 | 0.08 | 1.5 | 0.11 | 1.9 |
| QC 2 | 9.8 | 0.07 | 0.7 | 0.06 | 0.6 | 0.09 | 0.9 | 0.04 | 0.4 | 0.13 | 1.4 |
| QC 3 | 15.2 | 0.07 | 0.4 | 0.08 | 0.5 | 0.14 | 0.9 | 0.24 | 1.6 | 0.29 | 1.9 |

Table 3: Instrument 2 % (CV by Sample (NGSP))

Table 4: Instrument 3 (% CV by Sample (NGSP))

| | | | | Between- | | Between- | | | | Total | |
|-----------|--------|---------------|-----|----------|-----|----------|-----|-------------|-----|-----------|-----|
| Sample ID | HbA1c, | Repeatability | | Run | | Day | | Between-Lot | | Precision | |
| | % | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 5.2 | 0.06 | 1.2 | 0.00 | 0.0 | 0.06 | 1.2 | 0.05 | 0.9 | 0.10 | 1.9 |
| Patient 2 | 6.7 | 0.05 | 0.7 | 0.02 | 0.3 | 0.08 | 1.2 | 0.00 | 0.0 | 0.09 | 1.4 |
| Patient 3 | 8.3 | 0.05 | 0.6 | 0.03 | 0.4 | 0.11 | 1.3 | 0.04 | 0.5 | 0.13 | 1.6 |
| Patient 4 | 12.7 | 0.07 | 0.6 | 0.07 | 0.6 | 0.17 | 1.3 | 0.21 | 1.6 | 0.29 | 2.2 |
| Control 1 | 5.6 | 0.05 | 0.9 | 0.00 | 0.0 | 0.07 | 1.2 | 0.04 | 0.8 | 0.10 | 1.7 |
| Control 2 | 10.3 | 0.08 | 0.8 | 0.05 | 0.5 | 0.14 | 1.3 | 0.08 | 0.8 | 0.19 | 1.8 |
| QC 1 | 5.5 | 0.04 | 0.8 | 0.02 | 0.3 | 0.07 | 1.2 | 0.07 | 1.3 | 0.11 | 2.0 |
| QC 2 | 9.9 | 0.09 | 0.9 | 0.00 | 0.0 | 0.12 | 1.2 | 0.13 | 1.3 | 0.20 | 2.0 |
| QC 3 | 15.5 | 0.10 | 0.6 | 0.11 | 0.7 | 0.18 | 1.1 | 0.07 | 0.4 | 0.24 | 1.6 |

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| | | | | | | Between- | | Between- | | | Total | | |
|-----------|-------|---------------|-----|-------------|-----|----------|-----|-------------|-----|-------------|-------|-----------|-----|
| Sample ID | HbA1c | Repeatability | | Between-Run | | Day | | Instruments | | Between-Lot | | Precision | |
| | % | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 5.2 | 0.06 | 1.2 | 0.00 | 0.0 | 0.05 | 0.9 | 0.00 | 0.0 | 0.07 | 1.4 | 0.10 | 2.0 |
| Patient 2 | 6.7 | 0.05 | 0.8 | 0.01 | 0.2 | 0.07 | 1.0 | 0.05 | 0.8 | 0.03 | 0.5 | 0.11 | 1.6 |
| Patient 3 | 8.3 | 0.05 | 0.6 | 0.03 | 0.3 | 0.09 | 1.1 | 0.08 | 0.9 | 0.03 | 0.3 | 0.13 | 1.6 |
| Patient 4 | 12.7 | 0.08 | 0.6 | 0.07 | 0.6 | 0.16 | 1.2 | 0.00 | 0.0 | 0.21 | 1.7 | 0.28 | 2.2 |
| Control 1 | 5.6 | 0.06 | 1.0 | 0.00 | 0.0 | 0.06 | 1.0 | 0.00 | 0.0 | 0.05 | 0.9 | 0.10 | 1.7 |
| Control 2 | 10.3 | 0.08 | 0.7 | 0.04 | 0.4 | 0.12 | 1.1 | 0.14 | 1.3 | 0.10 | 1.0 | 0.22 | 2.2 |
| QC 1 | 5.5 | 0.05 | 0.8 | 0.00 | 0.1 | 0.06 | 1.0 | 0.00 | 0.0 | 0.08 | 1.4 | 0.11 | 2.0 |
| QC 2 | 9.9 | 0.10 | 1.0 | 0.00 | 0.0 | 0.10 | 1.0 | 0.05 | 0.5 | 0.09 | 0.9 | 0.17 | 1.8 |
| QC 3 | 15.4 | 0.09 | 0.6 | 0.12 | 0.8 | 0.14 | 0.9 | 0.20 | 1.3 | 0.17 | 1.1 | 0.34 | 2.2 |

Table 5: Instruments Combined (% CV by Sample (NGSP))

Table 6: Instrument 1 (% CV by Sample (IFCC Units- mmol/mol))

| Sample ID | HbA1c,<br>mmol/mol | Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Total Precision | |
|-----------|--------------------|---------------|-----|-------------|-----|-------------|-----|-------------|-----|-----------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 33 | 0.76 | 2.3 | 0.00 | 0.0 | 0.40 | 1.2 | 0.89 | 2.7 | 1.24 | 3.7 |
| Patient 2 | 50 | 0.76 | 1.5 | 0.00 | 0.0 | 0.81 | 1.6 | 0.54 | 1.1 | 1.23 | 2.5 |
| Patient 3 | 68 | 0.53 | 0.8 | 0.16 | 0.2 | 0.92 | 1.3 | 0.17 | 0.3 | 1.08 | 1.6 |
| Patient 4 | 116 | 1.01 | 0.9 | 0.65 | 0.6 | 1.85 | 1.6 | 2.31 | 2.0 | 3.19 | 2.8 |
| Control 1 | 38 | 0.77 | 2.0 | 0.00 | 0.0 | 0.56 | 1.5 | 0.53 | 1.4 | 1.09 | 2.9 |
| Control 2 | 91 | 1.08 | 1.2 | 0.00 | 0.0 | 1.31 | 1.4 | 1.34 | 1.5 | 2.16 | 2.4 |
| QC 1 | 37 | 0.63 | 1.7 | 0.00 | 0.0 | 0.53 | 1.4 | 0.93 | 2.5 | 1.25 | 3.4 |
| QC 2 | 86 | 1.46 | 1.7 | 0.00 | 0.0 | 0.96 | 1.1 | 0.61 | 0.7 | 1.85 | 2.2 |
| QC 3 | 147 | 1.25 | 0.9 | 1.67 | 1.1 | 0.92 | 0.6 | 1.90 | 1.3 | 2.96 | 2.0 |

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| Sample ID | HbA1c,<br>mmol/mol | Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Total Precision | |
|-----------|--------------------|---------------|-----|-------------|-----|-------------|-----|-------------|-----|-----------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 33 | 0.49 | 1.5 | 0.20 | 0.6 | 0.43 | 1.3 | 0.84 | 2.6 | 1.08 | 3.3 |
| Patient 2 | 49 | 0.36 | 0.7 | 0.42 | 0.9 | 0.58 | 1.2 | 0.24 | 0.5 | 0.83 | 1.7 |
| Patient 3 | 66 | 0.40 | 0.6 | 0.38 | 0.6 | 0.72 | 1.1 | 0.19 | 0.3 | 0.93 | 1.4 |
| Patient 4 | 113 | 0.63 | 0.6 | 0.88 | 0.8 | 1.47 | 1.3 | 2.27 | 2.0 | 2.91 | 2.6 |
| Control 1 | 37 | 0.46 | 1.2 | 0.19 | 0.5 | 0.61 | 1.6 | 0.70 | 1.9 | 1.05 | 2.8 |
| Control 2 | 87 | 0.45 | 0.5 | 0.66 | 0.8 | 0.89 | 1.0 | 0.96 | 1.1 | 1.54 | 1.8 |
| QC 1 | 37 | 0.38 | 1.0 | 0.21 | 0.6 | 0.61 | 1.7 | 0.91 | 2.5 | 1.18 | 3.2 |
| QC 2 | 84 | 0.72 | 0.9 | 0.67 | 0.8 | 1.00 | 1.2 | 0.43 | 0.5 | 1.46 | 1.7 |
| QC 3 | 142 | 0.73 | 0.5 | 0.83 | 0.6 | 1.52 | 1.1 | 2.58 | 1.8 | 3.19 | 2.2 |

Table 7: Instrument 2 (% CV by Sample (IFCC Units- mmol/mol))

Table 8: Instrument 3 (% CV by Sample (IFCC Units- mmol/mol))

| | HbA1c, | Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Total Precision | |
|-----------|----------|---------------|-----|-------------|-----|-------------|-----|-------------|-----|-----------------|-----|
| Sample ID | mmol/mol | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 33 | 0.70 | 2.1 | 0.00 | 0.0 | 0.66 | 2.0 | 0.51 | 1.6 | 1.09 | 3.3 |
| Patient 2 | 50 | 0.51 | 1.0 | 0.22 | 0.4 | 0.85 | 1.7 | 0.00 | 0.0 | 1.02 | 2.0 |
| Patient 3 | 68 | 0.56 | 0.8 | 0.33 | 0.5 | 1.20 | 1.8 | 0.46 | 0.7 | 1.44 | 2.1 |
| Patient 4 | 115 | 0.80 | 0.7 | 0.80 | 0.7 | 1.81 | 1.6 | 2.26 | 2.0 | 3.11 | 2.7 |
| Control 1 | 37 | 0.57 | 1.5 | 0.00 | 0.0 | 0.73 | 2.0 | 0.46 | 1.2 | 1.03 | 2.8 |
| Control 2 | 89 | 0.87 | 1.0 | 0.60 | 0.7 | 1.51 | 1.7 | 0.93 | 1.0 | 2.07 | 2.3 |
| QC 1 | 36 | 0.45 | 1.2 | 0.18 | 0.5 | 0.73 | 2.0 | 0.77 | 2.1 | 1.16 | 3.2 |
| QC 2 | 85 | 1.00 | 1.2 | 0.00 | 0.0 | 1.35 | 1.6 | 1.44 | 1.7 | 2.21 | 2.6 |
| QC 3 | 145 | 1.05 | 0.7 | 1.24 | 0.9 | 1.93 | 1.3 | 0.74 | 0.5 | 2.63 | 1.8 |

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

| Sample<br>ID | HbA1c,<br>mmol/mol | Repeatability | | Between-Run | | Between-<br>Day | | Between-<br>Instruments | | Between-<br>Lot | | Total<br>Precision | |
|--------------|--------------------|---------------|-----|-------------|-----|-----------------|-----|-------------------------|-----|-----------------|-----|--------------------|-----|
| | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Patient 1 | 33 | 0.66 | 2.0 | 0.00 | 0.0 | 0.51 | 1.5 | 0.00 | 0.0 | 0.77 | 2.3 | 1.13 | 3.4 |
| Patient 2 | 50 | 0.57 | 1.1 | 0.13 | 0.3 | 0.76 | 1.5 | 0.58 | 1.2 | 0.34 | 0.7 | 1.17 | 2.3 |
| Patient 3 | 67 | 0.50 | 0.7 | 0.31 | 0.5 | 0.96 | 1.4 | 0.84 | 1.3 | 0.30 | 0.4 | 1.44 | 2.1 |
| Patient 4 | 115 | 0.83 | 0.7 | 0.78 | 0.7 | 1.72 | 1.5 | 0.00 | 0.0 | 2.29 | 2.0 | 3.08 | 2.7 |
| Control 1 | 37 | 0.60 | 1.6 | 0.00 | 0.0 | 0.63 | 1.7 | 0.00 | 0.0 | 0.57 | 1.5 | 1.04 | 2.8 |
| Control 2 | 89 | 0.84 | 0.9 | 0.48 | 0.5 | 1.26 | 1.4 | 1.49 | 1.7 | 1.09 | 1.2 | 2.44 | 2.7 |
| QC 1 | 37 | 0.50 | 1.4 | 0.04 | 0.1 | 0.63 | 1.7 | 0.00 | 0.0 | 0.88 | 2.4 | 1.20 | 3.2 |
| QC 2 | 85 | 1.10 | 1.3 | 0.00 | 0.0 | 1.12 | 1.3 | 0.59 | 0.7 | 0.93 | 1.1 | 1.92 | 2.3 |
| QC 3 | 145 | 1.04 | 0.7 | 1.29 | 0.9 | 1.52 | 1.0 | 2.22 | 1.5 | 1.90 | 1.3 | 3.69 | 2.5 |

Table 9: Instruments Combined (% CV by Sample (IFCC Units- mmol/mol))

## b. Linearity

A linearity study was performed per CLSI EP06-A: Evaluation of the Linearity of Quantitative Measuring Procedures; A Statistical Approach. Linearity across the reportable range was performed using low (3.9%HbA1c) and high (18.8%HbA1c) EDTA whole blood patient samples. These samples were mixed together in varying ratios. The measured values were compared to the expected values based upon the dilution factor. Polynomial regression analysis (for first, second, and third order polynomials) were performed to determine the statistical significance of non-linearity. The higher order coefficients were found not to be significant and linearity was demonstrated.

% HbA1c (NGSP) using the D-10™ Hemoglobin A1c Program has demonstrated linearity from 3.9 – 18.8% HbA1c with the maximum measured difference of
± 0.1% between the predicted 1ª and 20 order results as shown in Table 10 below. Mmol/mol HbA1c (IFCC) has been demonstrated as linear from 19 – 182 mmol/mol with the maximum measured difference of ±1 mmol/mol as shown in Table 11 below.

## Table 10: Results of Linearity Study (NGSP %)

| Sample Pool | Predicted 1st Order | Predicted 2nd Order | Difference |
|-------------|---------------------|---------------------|------------|
| Level 1 | 3.7 | 3.8 | 0.1 |
| Level 2 | 5.2 | 5.3 | 0.1 |
| Level 3 | 6.8 | 6.8 | 0.0 |
| Level 4 | 8.3 | 8.2 | 0.1 |
| Level 5 | 9.8 | 9.7 | 0.1 |
| Level 6 | 11.3 | 11.2 | 0.1 |

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

| Level 7 | 12.8 | 12.7 | 0.1 |
|----------|------|------|-----|
| Level 8 | 14.3 | 14.3 | 0.0 |
| Level 9 | 15.8 | 15.8 | 0.0 |
| Level 10 | 17.3 | 17.3 | 0.0 |
| Level 11 | 18.8 | 18.9 | 0.1 |

#### Table 11: Results of Linearity Study (IFCC mmol/mol)

| Sample Pool | Predicted 1st Order | Predicted 2nd Order | Difference |
|-------------|---------------------|---------------------|------------|
| Level 1 | 17 | 18 | 1 |
| Level 2 | 34 | 34 | 0 |
| Level 3 | 50 | 50 | 0 |
| Level 4 | 67 | 67 | 0 |
| Level 5 | 83 | 83 | 0 |
| Level 6 | 100 | 99 | 1 |
| Level 7 | 116 | 116 | 0 |
| Level 8 | 133 | 133 | 0 |
| Level 9 | 149 | 149 | 0 |
| Level 10 | 166 | 166 | 0 |
| Level 11 | 182 | 183 | 1 |

#### Method Comparison ﻥ

A Method comparison study was performed per CLSI EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples. 128 variant-free whole blood K3 EDTA samples ranging from 3.9% to approximately 19.0 % (19 to approximately 184 mmol/mol) HbA1c were evaluated using the D-10™ Hemoglobin A1c Program on the D-10™ Hemoglobin Testing System. Samples were tested in a single determination over a 4 day period. The results were compared to testing performed at a secondary NGSP SRL reference laboratory using a cleared HPLC-based HbA1c assay. The distribution of samples spanned the measuring interval listed in Table 12.

| Hemoglobin A1c level | n | % Samples tested |
|----------------------|----|------------------|
| ≤ 5% | 4 | 3.1 |
| 5 - 6% | 19 | 14.8 |
| 6 – 6.5% | 33 | 25.8 |
| 6.5 - 7% | 33 | 25.8 |

## Table 12: Distribution of samples

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

| 7 – 8% | 20 | 15.6 |
|---------------|-----|------|
| 8 – 9% | 10 | 7.8 |
| > 9% | 9 | 7.0 |
| Total samples | 128 | 100 |

Linear, Deming (weighted) and Passing-Bablok regression analyses were performed for the D-10™ Hemoglobin A1c versus the NGSP SRL reference method. Deming (weighted), Passing-Bablok and Linear regression analyses were performed for the D-10™ Hemoglobin A1c on the D-10 Hemoglobin Testing System versus the reference G8 HPLC method are summarized in Table 13.

| Table 13: Summary of Method Comparison Results | | | | | |
|------------------------------------------------|--------|-----------------|-------------|-----------------|--------|
| | Slope | 95% CI | y-Intercept | 95% CI | R² |
| Linear | 0.9701 | 0.9587 – 0.9816 | 0.1801 | 0.0980 – 0.2622 | 0.9955 |
| Deming | 0.9722 | 0.9579 - 0.9866 | 0.1654 | 0.0637 - 0.2670 | 1.0000 |
| Passing-Bablok | 1.0000 | 1.0000 - 1.0000 | 0.0000 | 0.0000 - 0.0000 | 1.0000 |

## Table 13: Summary of Method Comparison Results

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

Image /page/14/Figure/0 description: This image is a scatter plot titled "Scatter Plot with Deming Fit". The plot shows the relationship between NGSP on the x-axis and D-10 Hemoglobin A1c Program on the y-axis. The plot includes a Deming fit line represented by the equation (0.17 + 0.97x), along with 95% confidence interval bands and an identity line for reference. The data points are clustered tightly around the Deming fit line, indicating a strong correlation between the two variables.

Figure 1: Scatter Plot using Deming Fit, %HbA1c, NGSP SRL vs. D-10 Hemoglobin A1c.

- (1) The following biases between D-10 Hemoglobin A1c versus NGSP SRL Method (Reference method) were observed in Table 14.

| % HbA1c – Decision<br>Level | Bias | % Bias |
|-----------------------------|-------|--------|
| 5.0±0.5 | -0.05 | -0.96 |
| 6.5±0.5 | 0.00 | 0.03 |
| 8.0±0.5 | -0.08 | -0.98 |
| 12.0±1.0 | -0.10 | -0.87 |

#### Table 14: Bias Estimation

Total Error Decision Levels

Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the reproducibility study, Total Error (TE) at four concentrations: (5.0 %, 6.5%, 8.0% and 12.0%) were calculated as follows: %TE=|%Bias|+1.96 *CV* (1 + %Bias). The results are presented in Table 15.

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

| % A1c - Decision Level | % Bias | % CV | % TE |
|------------------------|--------|------|------|
| 5.0 | 0.96 | 2.0 | 4.9 |
| 6.5 | 0.03 | 1.6 | 3.2 |
| 8.0 | 0.98 | 1.6 | 4.1 |
| 12.0 | 0.87 | 2.2 | 5.2 |

#### Table 15: Total Error Estimation

#### d. Traceability, Stability, Expected Values (calibrators)

The D-10 Hemoglobin A1c test standardization is traceable to the International Federation of Clinical Chemistry (IFCC) reference calibrators. The D-10 Hemoglobin A1c assay is NGSP certified. The NGSP certification expires in one year. See NGSP website for current certification at http://www.ngsp.org. The derived results of (%) from the NGSP correlation are calculated from the individual quantitative results for Hemogloblin A1c (HbA1c). The International Federation of Clinical Chemistry (IFCC) units of mmol/mol are calculated using the Master Equation NGSP (%) = 0.09148 x IFCC (mmol/mol) + 2.152. HbA1c results are provided to the customers using two different units: NGSP equivalent units (%) and IFCC equivalent units (mmol/mol).

## Calibrator Materials:

Value assignment for D-10™ Hemoglobin A1c Calibrators which are recommended for use with this device, were previously reviewed under 510(k) submission K031043.

## e. Analytical specificity:

- Endogenous Interference i.)
An Endogenous Interference study was performed per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of HbA1c of ~8.0%.

Coniugated bilirubin, unconiugated bilirubin and glucose, available in pure form, were obtained and stock solutions prepared at 10x the intended test concentration. The 10x stock solution of the test substance was pipetted into a low whole blood sample pool (at ~6.5% HbA1c) and a high whole blood sample pool (~8.0% HbA1c), making the test pool. Ten replicates of each pool prepared with the test and control samples were analyzed using the D-10TM Hemoglobin A1c on the D-10™Hemoglobin Testing System.

Rheumatoid factor, lipemia and total protein were not available as pure standards therefore serum samples with known concentration of these compounds were used. The test pool was prepared by mixing the serum sample known to have a high test substance concentration with a whole blood non-variant sample such that the concentration of test substance in the final mixture would be at the desired level. Ten replicates of each pool prepared with the test and control samples were analyzed using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System.

Significant interference was defined as a ± 7% chanqe in %HbA1c value from the control. Results in Table 16 showed no significant interference up to the stated concentrations.

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

| | Concentration | |
|------------------------|----------------------------|-------------|
| Endogenous substance | Conventional (US)<br>units | SI Units |
| Lipemia (Intralipid) | 6000 mg/dL | 60 g/L |
| Conjugated bilirubin | 60 mg/dL | 712 µmol/L |
| Unconjugated bilirubin | 60 mg/dL | 1026 µmol/L |
| Glucose | 2000 mg/dL | 111 mmol/L |
| Rheumatoid factor | 750 IU/mL | 750 kIU/mL |
| Total protein | 21 g/dL | 210 g/L |

#### Table 16: Endogenous Interference Study Results

#### ii.) Drug Interference:

A Drug Interference study was performed based per CLSI EP07-A2, Interference Testing in Clinical Chemistry. Two EDTA whole blood sample pools were evaluated using a low level whole blood sample with a concentration ~6.5%HbA1c and a high level whole blood sample with a concentration of ~8.0%HbA1c. Test samples were prepared by spiking each drug at the interferent concentration shown in Table 18. Ten replicates of each drug prepared with the test and control samples were analyzed using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System.

Significant interference was defined as a more than ± 7% change in %HbA1c value from the control. No significant interference was observed at therapeutic levels up to the stated concentrations in Table 17 on the following page.

{17}------------------------------------------------

| Potential Drug<br>Interferent | Highest Level Tested showing no Significant<br>Interference | |
|-------------------------------|-------------------------------------------------------------|--------------|
| | Conventional (US)<br>units | SI units |
| Acetylcysteine | 166 mg/dL | 10.2 mmol/L |
| Ampicillin-Na | 1000 mg/dL | 28.65 mmol/L |
| Ascorbic acid | 300 mg/dL | 17.05 mmol/L |
| Cefoxitin | 2500 mg/dL | 58.55 mmol/L |
| Heparin | 5000 U/L | 5000 U/L |
| Levodopa | 20 mg/dL | 1015 µmol/L |
| Methyldopa | 20 mg/dL | 948 µmol/L |
| Metronidazole | 200 mg/dL | 11.7 mmol/L |
| Doxycyclin | 50 mg/dL | 1124 µmol/L |
| Acetylsalicylic acid | 1000 mg/dL | 55.51 mmol/L |
| Rifampicin | 64 mg/L | 78 µmol/L |
| Cyclosporine | 5 mg/L | 4 µmol/L |
| Acetaminophen | 200 mg/L | 1323 µmol/L |
| Ibuprofen | 500 mg/L | 2427 µmol/L |
| Theophylline | 100 mg/L | 556 µmol/L |
| Phenylbutazone | 400 mg/L | 1299 µmol/L |

#### Table 17: Drug Interference Study Results

#### iii.) Cross Reactivity with Hemoglobin Derivatives:

A Hemoglobin Derivatives Interference study was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. Potential interference from Acetylated hemogloblin (Hb), Carbamylated hemoglobin (Hb) and Labile HbA1c were evaluated using a low level whole blood EDTA sample with a concentration ~6.5%HbA1c and a high level whole blood EDTA sample with a concentration of

{18}------------------------------------------------

~8.0% HbA1c. The potentially interfering hemoglobin derivatives were spiked into the low and high level blood samples and each sample was analyzed using ten replicates each in the same analytical run on the D-10™ Hemoglobin Testing System with the D-10™Hemoglobin A1c.

Significant interference was defined as more than a ±7% change in HbA1c value from the control. The test result conclusions are as follows:

- . Acetylated Hb- up to 49 mg/dL does not interfere with this assay.
- . Carbamylated Hb - up to 3.5% (or 10 mg/dL potassium cyanate) does not interfere with this assay.
- Labile A1c- up to 6% (or 1000 mg/dL glucose) does not interfere with . this assay.

Results showed there was no cross reactivity with these substances at physiological levels.

- iv.) Hemoglobin Variant Study:
A Hemoglobin Variant study was performed using a panel of normal and diabetic whole blood EDTA patent variant samples known to contain hemoglobin variants S, C, E, D, A2 and F. Testing of the samples containing hemoglobin variants S, C, E, D, A2 and F were performed in duplicate using the D-10™ Hemoglobin A1c on the D-10™ Hemoglobin Testing System and compared to results obtained by a NGSP reference method that has been demonstrated to be free from the hemoglobin interferent. Table 18 contains the number of samples, range of samples and concentration of samples used in the Hemoglobin Variant Study. Table 19 contains the results for the Hemoglobin Variant study bias.

| Table 10: Variant samples used in Hemoglobin Variant Study | | | |
|------------------------------------------------------------|----|---------------------|-----------------|
| Hemoglobin | n | Range in % Abnormal | Range in %HbA1c |
| Variant | | Variant | Concentration |
| HbS | 22 | 31 - 43 | 5.6 - 11.5 |
| HbC | 20 | 31 - 40 | 5.0 - 10.7 |
| HbD | 22 | 35 - 43 | 5.8 - 10.0 |
| HbE | 23 | 21 - 32 | 5.9 - 11.6 |
| HbA2 | 20 | 5.0 - 6.2 | 5.0 - 14.5 |
| HbF | 24 | 3.3 - 32.6 | 4.7 - 14.4 |

#### Table 18: Variant samples used in Hemoglobin Variant Study

#### Table 19: Hemoqlobin Variant Study Bias Results

| Hemoglobin<br>Variant | Relative % Bias to Comparative Method | |
|-----------------------|----------------------------------------------|----------------------------------------------|
| | Relative %Bias (Range of %Bias) for<br>HbA1c | Relative %Bias (Range of %Bias)<br>for HbA1c |
| | ~6.5% | ~9.0% |
| HbS | 1.1 (-4.7 to 4.9) | 0.2 (-6.1 to5.3) |
| HbC | 2.6 (1.4 to 5.9) | -0.9 (-2.8 to 0.9) |
| HbD | -0.6 (-3.1 to 4.7) | 1.7 (-3.6 to 5.1) |
| HbE | 0.5 (-3.3 to 6.2) | 2.8 (1.2 to 5.2) |
| HbA2 | 0.6 (-1.8 to 1.8) | -1.7 (-2.8 to -0.7) |
| HbF | 3.1 (-1.5 to 8.8) | -0.6 (-2.5 to 4.8) |

{19}------------------------------------------------

This device has significant positive interference with fetal hemoglobin (HbF). HbA1c results are invalid for patients with abnormal amounts of HbF including those with known Hereditary Persistence of Fetal Hemoglobin.

Hemoglobin F concentrations up to 10% do not interfere with the test. Any sample with HbF>10% may result in higher than expected HbA1c values. Any sample with HbF >5% should be suspected of having a hemoglobinopathy.

No significant interference was observed for HbC (≤ 40%), HbD (≤ 43%), HbS (≤ 43%), HbE (≤ 32%),HbA2 (≤ 6%), and HbF(≤ 10%) at the concentrations tested in this study.

#### f. Matrix comparison

The data supports the use of the following blood collection tubes with the D-10™ Hemoglobin A1c test in Table 20.

| Table 20: Anticoagulant | | | |
|-------------------------|--|--|--|
| 1 นิ้มเป็นครั้ง | | | |
| Ko-EDTA | | | |
| Kz.EDTA | | | |

## g. Expected Values/Reference Range

Hemoglobin A1c expected values range was cited from American Diabetes Association Standards of Medical Care in Diabetes 2014, 37 (Supplement 1) and American Diabetes Association. Standards of Medical Care in Diabetes - 2016 are presented in Table 21.

| Table 21: Hemoqlobin A1c Expected Values | | |
|------------------------------------------|--|--|
| | | |

| Hemoglobin A1c | | Suggested Diagnosis |
|----------------|---------------|---------------------|
| NGSP % | IFCC mmol/mol | |
| >6.5 | >47 | Diabetic |
| 5.7 - 6.4 | 39-46 | Pre-Diabetic |
| <5.7 | <39 | Non-Diabetic |

#### Conclusion:

The information and data in this 510(k) document demonstrate that the D-10™ Hemoglobin A1c Program as performed on the D-10™Hemoglobin Testing System is an accurate, reliable, precise test that correlates well with current cleared methods and NGSP standardized testing for the quantitation of HbA1c. The contents of this submission demonstrates that the D-10™ Hemoglobin A1c Program as performed on the D-10™ Hemoglobin Testing System is substantially equivalent to its predicate device, VARIANT II TURBO HbA1c Kit -2.0 and, therefore, safe and effective for its intended use. The performance criteria as stipulated by the Special Controls requirements for HbA1c systems that diagnose diabetes have clearly been met. The D-10™ Hemoglobin A1c Program must be found to be substantially equivalent to the predicate and, therefore, cleared by the agency for the intended use requested.