CAPILLARYS Hb A1c

K171861 · Sebia · PDJ · Feb 7, 2018 · Clinical Chemistry

Device Facts

• Record ID: K171861
• Device Name: CAPILLARYS Hb A1c
• Applicant: Sebia
• Product Code: PDJ · Clinical Chemistry
• Decision Date: Feb 7, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1373
• Device Class: Class 2

Indications for Use

The CAPILLARYS 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 2 FLEX-PIERCING 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 CAPILLARYS Hb A1c kit is intended for in vitro Diagnostic Use Only.

Device Story

The CAPILLARYS Hb A1c system uses capillary electrophoresis in an alkaline buffer to separate and quantify the HbA1c glycated fraction of hemoglobin from venous whole blood. The CAPILLARYS 2 FLEX-PIERCING instrument performs 8 simultaneous analyses; samples are diluted, aspirated, and subjected to high-voltage protein separation. Hemoglobins are detected directly at the cathodic end via 415 nm absorbance. The system provides HbA1c concentrations in NGSP (%) and IFCC (mmol/mol) units. Used in clinical laboratories, the device aids physicians in diagnosing diabetes, identifying at-risk patients, and monitoring long-term glucose control. The high-resolution procedure allows quantification in the presence of labile HbA1c, carbamylated/acetylated hemoglobins, and major variants (HbA2, S, C, D, E, F). Results assist clinicians in assessing patient glycemic status and adjusting therapeutic interventions.

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP5-A3 over 20 days using 3 lots, demonstrating total reproducibility CVs between 1.2% and 3.4%. Linearity confirmed across 4.4%–16.6% (NGSP) range per CLSI EP06-A. Method comparison study (n=150) against a cleared HPLC method showed high correlation (r=0.999) and low bias. Interference studies (CLSI EP07-A2) confirmed no significant interference from common endogenous factors, drugs, or hemoglobin variants (S, C, D, E, A2, F) up to specified concentrations.

Technological Characteristics

Capillary electrophoresis in free solution; alkaline buffer; silica capillaries; 415 nm absorbance detection. Dimensions/form factor: CAPILLARYS 2 FLEX-PIERCING instrument. Connectivity: Standalone. Software: PHORESIS software. Reagents: Buffer, hemolysing solution, wash solution. Standardization: Traceable to IFCC reference calibrators; NGSP certified.

Indications for Use

Indicated for use in venous whole blood samples for the diagnosis of diabetes, identification of patients at risk for diabetes, and monitoring long-term blood glucose control in patients with diabetes mellitus. Contraindicated for point-of-care use, daily glucose monitoring, or as a replacement for home testing. Not for use in pediatric patients, pregnant women, patients with Type 1 diabetes, or patients with conditions affecting red blood cell lifespan (e.g., hemolytic anemia, hemoglobinopathies, severe hepatic/renal disease, recent blood loss/transfusion). Not for diagnosing gestational diabetes.

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-723G8 (k131580)

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION MEMORANDUM ASSAY ONLY TEMPLATE

A. 510(k) Number:
k171861

B. Purpose for Submission:
Addition of a diagnostic claim to a previously cleared device

C. Measurand:
Hemoglobin A1c

D. Type of Test:
Capillary electrophoresis

E. Applicant:
Sebia, Inc.

F. Proprietary and Established Names:
CAPILLARYS Hb A1c

G. Regulatory Information:
| Product Code | Regulation Name | Classification | Regulation Section | Panel |
| --- | --- | --- | --- | --- |
| PDJ | Hemoglobin A1c Test System | Class II | 21 CFR 862.1373 | Clinical Chemistry (75) |

H. Intended Use:
1. Intended use(s):
See indications for use below.

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2. Indication(s) for use:

The CAPILLARYS 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 2 FLEX-PIERCING 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 CAPILLARYS Hb A1c kit is intended for in vitro Diagnostic Use Only.

3. Special conditions for use statement(s):

For In Vitro Diagnostic Use
Prescription Use Only

The CAPILLARYS Hb A1c kit should not be used:
- for point-of-care use
- in monitoring daily glucose control
- to replace daily home testing of urine and blood glucose levels
- to replace glucose testing in pediatric patients, pregnant women, or patients with Type 1 diabetes.
- for analyzing samples from patients with total hemoglobin levels of less than 2.9 or greater than 30.5 g/dL and any hemoglobinopathies that may interfere.
- to diagnose diabetes during pregnancy or to diagnose gestational diabetes. HbA1c reflects the average blood glucose levels over the preceding 3 months (the average life of a red blood cell), and therefore may be falsely low during pregnancy or any other condition associated with recent onset of hyperglycemia and/or decreased red cell survival,
- to diagnose diabetes in patients with the following conditions:
- Any condition that alters the life span of the red blood cells, including recent blood loss, transfusion, significant iron deficiency, hemolytic anemia (including hereditary spherocytosis) or other hemolytic diseases, hemoglobinopathies and thalassemias, as the altered red blood cell turnover interferes with the relationship between mean blood glucose and HbA1c values, malignancies or severe chronic hepatic and renal disease.
- In cases of rapidly evolving Type 1 diabetes, the increase of HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions, diabetes mellitus must be diagnosed based on plasma glucose concentration and/or the typical clinical symptoms.

4. Special instrument requirements:

CAPILLARYS 2 FLEX-PIERCING instrument

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I. Device Description:

The CAPILLARYS Hb A1c kit is to be used with the CAPILLARYS 2 FLEX-PIERCING instrument. The CAPILLARYS Hb A1c kit contains a ready to use buffer solution pH 9.4 ± 0.5 (2 vials, 700 ml each), a ready to use hemolysing solution (1 vial, 700 ml), Wash solution (1 vial, 75 ml), Green Dilution segments (1 pack of 90) and filters (4 filters per kit).

J. Substantial Equivalence Information:

1. Predicate device name(s):

TOSOH G8 Automated Glycohemoglobin Analyzer HLC-723G8

2. Predicate 510(k) number(s):

k131580

3. Comparison with predicate:

| Similarities and Differences | | |
| --- | --- | --- |
| Item | CAPILLARYS Hb A1c Kit (Candidate Device) | TOSOH G8 Automated Glycohemoglobin Analyzer HLC-723G8 k131580 (Predicate Device) |
| Intended Use | The CAPILLARYS Hb A1c kit is intended to be 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. | Same |
| Specimen Type | Venous whole blood (K₂EDTA and K₃EDTA) | Same |
| Standardization | Traceable to the Diabetes Control and Complications Trial (DCCT) reference method and IFCC. Certified via the National Glycohemoglobin Standardization Program (NGSP) | Same |
| Assay Principle | Capillary electrophoresis | Ion-exchange HPLC |
| Measuring Range | 4.4-16.6% (NGSP)
24-158 mmol/mol (IFCC) | 4.0-16.9% |

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K. Standard/Guidance Document Referenced (if applicable):

Clinical and Laboratory Standards Institute (CLSI) EP05-A3: Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline—Third Edition

CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline

CLSI EP07-A2: Interference Testing in Clinical Chemistry; Approved Guideline—Second Edition

CLSI EP09-A2: Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline—Second Edition

L. Test Principle:

The CAPILLARYS Hb A1c kit uses the principle of capillary electrophoresis in free solution. With this technique, charged molecules of hemoglobin are separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation of the glycated HbA1c fraction from other hemoglobins occurs according to the electrolyte pH and electroosmotic flow.

The CAPILLARYS 2 FLEX-PIERCING instrument has silica capillaries functioning in parallel allowing 8 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 HbA1c is made at the cathodic end of the capillary at 415 nm, which is the absorbance wave length specific to HbA1c. Before each run, the capillaries are washed with a wash solution and prepared for the next analysis with buffer.

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

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision was evaluated for the CAPILLARYS Hb A1c kit based on CLSI EP05-A3 using four K₂EDTA venous whole blood samples with targeted HbA1c concentrations (5.1%, 6.4%, 8.2%, and 12.2%), two quality control solutions (5.1% and 8%), and two calibrators (5.6% and 10.1%). Samples were analyzed on three CAPILLARYS 2 FLEX-PIERCING instruments using three lots of reagents. Each sample was analyzed in duplicate on two capillaries per run, two runs per day for 20 days. The results are shown below:

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# Results in NGSP units (%HbA1c)

Instruments Combined (%CV by Sample, NGSP % HbA1c)

| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Between instrument | Total |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 5.1 | 1.1% | 1.2% | 0.0% | 0.6% | 0.5% | 0.0% | 1.8% |
| Sample 2 | 6.4 | 1.1% | 0.6% | 0.0% | 0.4% | 0.6% | 0.0% | 1.4% |
| Sample 3 | 8.2 | 0.9% | 0.7% | 0.0% | 0.3% | 0.4% | 0.0% | 1.2% |
| Sample 4 | 12.2 | 0.7% | 0.6% | 0.0% | 0.4% | 0.9% | 0.0% | 1.3% |
| Control 1 | 5.1 | 1.1% | 1.1% | 0.0% | 0.7% | 1.0% | 0.3% | 2.0% |
| Control 2 | 8.0 | 0.9% | 1.1% | 0.0% | 0.9% | 0.2% | 1.1% | 2.0% |
| Calibrator 1 | 5.6 | 1.3% | 0.7% | 0.0% | 0.5% | 0.9% | 0.0% | 1.8% |
| Calibrator 2 | 10.1 | 0.8% | 0.6% | 0.0% | 0.2% | 0.7% | 0.0% | 1.3% |

Instrument 1 (%CV by Sample, NGSP % HbA1c)

| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 5.1 | 1.3% | 0.7% | 0.0% | 0.5% | 0.7% | 1.7% |
| Sample 2 | 6.4 | 1.1% | 0.5% | 0.0% | 0.4% | 0.5% | 1.4% |
| Sample 3 | 8.2 | 0.9% | 0.6% | 0.0% | 0.4% | 0.5% | 1.3% |
| Sample 4 | 12.2 | 0.7% | 0.6% | 0.0% | 0.4% | 0.8% | 1.3% |
| Control 1 | 5.1 | 1.2% | 0.7% | 0.0% | 0.7% | 0.8% | 1.8% |
| Control 2 | 8.0 | 0.9% | 1.5% | 0.0% | 0.4% | 0.3% | 1.8% |
| Calibrator 1 | 5.6 | 1.2% | 0.7% | 0.0% | 0.7% | 0.7% | 1.7% |
| Calibrator 2 | 10.1 | 0.9% | 0.7% | 0.0% | 0.2% | 0.7% | 1.3% |

Instrument 2 (%CV by Sample, NGSP % HbA1c)

| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 5.1 | 1.0% | 0.4% | 0.3% | 0.5% | 0.2% | 1.2% |
| Sample 2 | 6.4 | 1.1% | 0.6% | 0.0% | 0.4% | 0.8% | 1.5% |
| Sample 3 | 8.2 | 0.8% | 0.3% | 0.2% | 0.3% | 0.6% | 1.1% |
| Sample 4 | 12.2 | 0.7% | 0.5% | 0.0% | 0.3% | 1.0% | 1.3% |
| Control 1 | 5.1 | 1.1% | 1.0% | 0.0% | 0.5% | 1.1% | 1.9% |
| Control 2 | 8.0 | 0.7% | 1.1% | 0.0% | 0.7% | 0.3% | 1.5% |
| Calibrator 1 | 5.6 | 1.3% | 0.6% | 0.0% | 0.4% | 1.1% | 1.8% |
| Calibrator 2 | 10.1 | 0.8% | 0.5% | 0.3% | 0.2% | 0.7% | 1.2% |

Instrument 3 (%CV by Sample, NGSP % HbA1c)

| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 5.1 | 1.1% | 1.9% | 0.0% | 0.7% | 0.6% | 2.4% |
| Sample 2 | 6.4 | 1.2% | 0.6% | 0.0% | 0.3% | 0.3% | 1.4% |
| Sample 3 | 8.2 | 0.9% | 0.9% | 0.0% | 0.2% | 0.0% | 1.3% |

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| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 4 | 12.2 | 0.7% | 0.8% | 0.0% | 0.4% | 0.7% | 1.3% |
| Control 1 | 5.1 | 1.2% | 1.3% | 0.0% | 0.7% | 1.0% | 2.1% |
| Control 2 | 8.0 | 1.0% | 0.6% | 0.0% | 1.3% | 0.0% | 1.7% |
| Calibrator 1 | 5.6 | 1.4% | 0.9% | 0.0% | 0.5% | 0.9% | 1.9% |
| Calibrator 2 | 10.1 | 0.9% | 0.7% | 0.0% | 0.2% | 0.5% | 1.2% |

## Results in IFCC units (mmol/mol)

Instruments Combined (%CV by Sample, IFCC units)

| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Between instrument | Total |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 32 | 1.9% | 2.1% | 0.0% | 0.8% | 1.1% | 0.0% | 3.1% |
| Sample 2 | 46 | 1.7% | 0.9% | 0.0% | 0.6% | 0.8% | 0.0% | 2.1% |
| Sample 3 | 66 | 1.2% | 0.9% | 0.0% | 0.5% | 0.6% | 0.0% | 1.6% |
| Sample 4 | 109 | 0.8% | 0.8% | 0.0% | 0.5% | 1.1% | 0.0% | 1.6% |
| Control 1 | 33 | 2.0% | 1.8% | 0.0% | 1.1% | 1.8% | 0.1% | 3.4% |
| Control 2 | 63 | 1.2% | 1.6% | 0.0% | 1.2% | 0.3% | 1.4% | 2.7% |
| Calibrator 1 | 37 | 2.0% | 1.1% | 0.0% | 0.8% | 1.2% | 0.0% | 2.7% |
| Calibrator 2 | 87 | 1.1% | 0.8% | 0.0% | 0.3% | 0.7% | 0.0% | 1.5% |

Instrument 1 (%CV by Sample, IFCC units)

| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 32 | 1.9% | 1.5% | 0.0% | 0.7% | 1.3% | 2.8% |
| Sample 2 | 46 | 1.7% | 0.8% | 0.0% | 0.5% | 0.8% | 2.1% |
| Sample 3 | 66 | 1.2% | 0.8% | 0.0% | 0.7% | 0.6% | 1.6% |
| Sample 4 | 109 | 0.8% | 0.7% | 0.0% | 0.5% | 1.0% | 1.6% |
| Control 1 | 33 | 2.0% | 1.5% | 0.0% | 1.2% | 1.6% | 3.2% |
| Control 2 | 63 | 1.2% | 2.2% | 0.0% | 0.6% | 0.5% | 2.6% |
| Calibrator 1 | 37 | 2.1% | 0.8% | 0.0% | 1.1% | 0.8% | 2.7% |
| Calibrator 2 | 87 | 1.1% | 0.9% | 0.0% | 0.2% | 0.7% | 1.6% |

Instrument 2 (%CV by Sample, IFCC units)

| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 32 | 1.9% | 0.8% | 0.0% | 0.7% | 0.7% | 2.3% |
| Sample 2 | 46 | 1.5% | 0.8% | 0.0% | 0.8% | 1.0% | 2.1% |
| Sample 3 | 66 | 1.1% | 0.5% | 0.3% | 0.4% | 0.8% | 1.5% |
| Sample 4 | 109 | 0.8% | 0.7% | 0.0% | 0.4% | 1.3% | 1.7% |
| Control 1 | 33 | 1.9% | 1.4% | 0.0% | 0.8% | 1.8% | 0.1% |
| Control 2 | 63 | 0.9% | 1.4% | 0.0% | 1.0% | 0.3% | 1.4% |

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| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Calibrator 1 | 37 | 1.8% | 0.8% | 0.0% | 0.7% | 1.5% | 2.6% |
| Calibrator 2 | 87 | 1.0% | 0.5% | 0.3% | 0.4% | 0.9% | 1.5% |

Instrument 3 (%CV by Sample, IFCC units)

| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample 1 | 39 | 1.9% | 3.2% | 0.0% | 1.0% | 1.1% | 3.9% |
| Sample 2 | 46 | 1.7% | 1.0% | 0.3% | 0.5% | 0.5% | 2.1% |
| Sample 3 | 66 | 1.3% | 1.2% | 0.0% | 0.3% | 0.0% | 1.8% |
| Sample 4 | 109 | 0.8% | 1.0% | 0.0% | 0.5% | 0.9% | 1.7% |
| Control 1 | 33 | 2.1% | 2.2% | 0.0% | 1.3% | 1.7% | 3.7% |
| Control 2 | 63 | 1.3% | 0.8% | 0.0% | 1.8% | 0.0% | 2.4% |
| Calibrator 1 | 37 | 2.1% | 1.5% | 0.0% | 0.7% | 1.1% | 2.9% |
| Calibrator 2 | 87 | 1.2% | 0.8% | 0.0% | 0.2% | 0.5% | 1.5% |

b. Linearity/assay reportable range:

The linearity of the CAPILLARYS Hb A1c kit was evaluated based on CLSI EP06-A. Two blood samples, including a normal venous K₂EDTA whole blood sample with HbA1c concentration at 4.4% (24 mmol/mol) and an elevated HbA1c level venous whole blood sample with HbA1c concentration at 16.7% (159 mmol/mol) were mixed within different proportions (5.3%, 6.3%, 7.3%, 8.4%, 9.6%, 10.8%, 12.1%, 13.6%, and 15.1%) and the mixtures were electrophoresed with the CAPILLARYS Hb A1c kit. For each mixture, samples were analyzed in triplicate.

The following table summarizes the linear regression results for both NGSP (%HbA1c) and IFCC units (mmol/mol).

| Units | Slope | y-intercept | Correlation coefficient (r) |
| --- | --- | --- | --- |
| NGSP (%HbA1c) | 1.005 | -0.124 | 1.000 |
| IFCC (mmol/mol) | 1.006 | -1.253 | 1.000 |

The study supports the claimed reportable range of the device from 4.4% to 16.6% (24-158 mmol/mol) HbA1c.

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Traceability:

The CAPILLARYS Hb A1c kit is traceable to the International Federation of Clinical Chemistry (IFCC) reference calibrators. The CAPILLARYS Hb A1c kit is certified with the National Glycohemoglobin Standardization Program (NGSP). The NGSP certification expires in one year. See NGSP website for current certification at http://www.ngsp.org.

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The derived result of the ratio (\%) from the NGSP correlation is calculated from the individual quantitative results for Hemoglobin A1c (HbA1c). The International Federation of Clinical Chemistry (IFCC) units of mmol/mol are calculated using the Master Equation: $\mathrm{IFCC} = (\mathrm{NGSP} - 2.15) / 0.092$ . Two different units are provided to the customers: NGSP equivalent units (\%) and IFCC equivalent units (mmol/mol).

# d. Detection limit:

The claimed measuring range of 4.4 to $16.6\%$ for the CAPILLARYS Hb A1c kit is based on linearity, see M.1.b.

# e. Analytical specificity:

# i. Endogenous Interference

Interference studies were performed to assess endogenous substances that could interfere with the CAPILLARYS Hb A1c kit. The interfering substances were evaluated in two venous whole blood $\mathrm{K}_2\mathrm{EDTA}$ samples with one sample close to the cut-off value, $\sim 6.3\%$ (45 mmol/mol) HbA1c, and one sample with high levels of HbA1c, $\sim 8.3\%$ (67 mmol/mol) HbA1c. Test samples were prepared by spiking each endogenous substance at a single dose to simulate the expected dose into the two HbA1c samples. Test samples were assayed in replicates of 10 and the mean concentration for each test sample was compared to the mean concentration of 10 replicates of the control sample. The sponsor defined significant interference as greater than or equal to $7\%$ mean relative deviation between the test and control sample.

The highest concentration of endogenous substances tested that show non-significant interference are summarized in the table below:

| Potential interfering substance | Highest concentration tested without significant interference |
| --- | --- |
| Ascorbic Acid | 300 mg/dL |
| Conjugated bilirubin | 60 mg/dL |
| Unconjugated bilirubin | 60 mg/dL |
| Rheumatoid factor | 1076 IU/mL |
| Triglycerides | 2.89 g/dL |
| Urea | 265 mg/dL |
| D-glucose | 1000 mg/dL |
| Total Protein | 149.5 g/L |

# ii. Exogenous Interference

Interference studies were performed to assess various exogenous substances that could interfere with the CAPILLARYS Hb A1c kit. The interfering substances were

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evaluated in two venous whole blood $\mathrm{K}_2\mathrm{EDTA}$ samples with one sample close to the cut-off value, $6.3\%$ (45 mmol/mol) HbA1c, and one sample with high levels of HbA1c, $8.3\%$ (67 mmol/mol) HbA1c. Test samples were prepared by spiking each drug at a single dose to simulate the maximum expected dose into the two HbA1c samples. Test samples were assayed in replicates of 10 and the mean concentration for each test sample was compared to the mean concentration of 10 replicates of the control sample. The sponsor defined significant interference as greater than or equal to $7\%$ mean relative deviation between the test and control samples.

The highest concentrations of exogenous substances tested that show no significant interference are summarized in the table below:

| Potential interfering substance | Highest concentration tested without significant interference |
| --- | --- |
| Acetaminophen | 20 mg/dL (1325 μM) |
| Acetylcysteine | 200 mg/dL (12.3 mM) |
| Acetylsalicylic acid | 1000 mg/dL (55,56 mM) |
| Ampicillin-Na | 1000 mg/dL (28653 μM) |
| Cefoxitin | 2500 mg/dL (58548 μM) |
| Cyclosporine | 0.5 mg/dL |
| Doxycycline | 50 mg/dL (1123.6 μM) |
| Glybenclamide | 3 mg/dL |
| Heparin | 5000 U/L |
| Ibuprofen | 50 mg/dL (2427 μM) |
| Levodopa | 40 mg/dL |
| Metformin | 5 mg/dL (387 μM) |
| Methyldopa | 40 mg/dL (1896 μM) |
| Metronidazole | 200 mg/dL (11696 μM) |
| Phenylbutazone | 40 mg/dL |
| Rifampicin | 7 mg/dL (85.1 μM) |
| Theophylline | 10 mg/dL (556 μM) |

# iii. Cross Reactivity with Hemoglobin Derivatives

To study interference from carbamylated hemoglobin, two $\mathrm{K}_2\mathrm{EDTA}$ whole blood patient samples with HbA1c concentrations at $\sim 6.3\%$ (46 mmol/mol) and $\sim 8.4\%$ (69 mmol/mol) were split into two aliquots. One aliquot, at each HbA1c level, was spiked with 1 mmol/L of potassium cyanate. The other aliquot, at each HbA1c level, was not spiked and served as a control sample. Samples were then analyzed on the CAPILLARYS 2 FLEX-PIERCING instrument using the CAPILLARYS Hb A1c kit. Samples were analyzed in replicates of 10. The sponsor's definition of nonsignificant interference is $\leq 7\%$ mean relative deviation between the test and the control sample. The sponsor concluded that carbamylated HbA1c up to $810~\mathrm{mg / dL}$ (7.2%) does not interfere with this assay.

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To study interference from labile hemoglobin, two K₂EDTA whole blood patient samples with HbA1c concentrations at ~6.2% (44 mmol/mol) and ~8.1% (65 mmol/mol) were split into two aliquots. One aliquot, at each HbA1c level, was spiked with glucose (50 g/L). The other aliquot, at each HbA1c level, was not spiked and served as a control sample. Samples were then analyzed on the CAPILLARYS 2 FLEX-PIERCING instrument using the CAPILLARYS Hb A1c kit. Samples were analyzed in replicates of 10. The sponsor’s definition of non-significant interference is ≤7% mean relative deviation between the test and the control sample. The sponsor concluded that labile HbA1c up to 1970 mg/dL (18.4%) does not interfere with this assay.

To study interference from acetylated hemoglobin, two K₂EDTA whole blood samples with HbA1c concentrations at ~6.2% (45 mmol/mol) and ~8.5% (69 mmol/mol) were split into two aliquots. One aliquot, at each HbA1c level, was spiked with acetylsalicylic acid (10 mmol/L). The other aliquot, at each HbA1c level, was not spiked and served as a control sample. Samples were then analyzed on the CAPILLARYS 2 FLEX-PIERCING instrument using the CAPILLARYS Hb A1c kit. Samples were analyzed in replicates of 10. The sponsor’s definition of non-significant interference is ≤7% mean relative deviation between the test and the control sample. The sponsor concluded that acetylated HbA1c up to 420 mg/dL (3.6%) does not interfere with this assay.

To study interference from HbA1a+b, two K₂EDTA whole blood samples with HbA1c concentrations at ~6.5% and (48 mmol/mol) ~8.4% (68 mmol/mol) were split into two aliquots. One aliquot, at each HbA1c level, was spiked with HbA1a+b (20 mg/dL). The other aliquot, at each HbA1c level, was without HbA1a+b and served as a control sample. Samples were then analyzed on the CAPILLARYS 2 FLEX-PIERCING instrument using the CAPILLARYS Hb A1c kit. Samples were analyzed in replicates of 10. The sponsor’s definition of non-significant interference is ≤7% mean relative deviation between the test and the control sample. The sponsor concluded that HbA1a+b up to ≤20 mg/dL does not interfere with this assay.

To study interference from glycated albumin, two K₂EDTA whole blood samples with HbA1c concentrations at ~6.1% and (45 mmol/mol) ~8.7% (68 mmol/mol) were split into two aliquots. One aliquot, at each HbA1c level, was spiked with glycated albumin (220 mg/dL). The other aliquot, at each HbA1c level, was without glycated albumin and served as a control sample. Samples were then analyzed on the CAPILLARYS 2 FLEX-PIERCING instrument using the CAPILLARYS Hb A1c kit. Samples were analyzed in replicates of 10. The sponsor’s definition of non-significant interference is ≤7% mean relative deviation between the test and the control sample. The sponsor concluded that glycated albumin up to 220 mg/dL does not interfere with this assay.

Concentration at which no significant interference (≤7%) was observed for each hemoglobin derivative is presented below:

10

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| Interfering Factor | Highest concentration tested without significant interference |
| --- | --- |
| Carbamylated hemoglobin | 810 mg/dL |
| Labile HbA1c | 1970 mg/dL |
| Acetylated hemoglobin | 420 mg/dL |
| HbA1a+b | 20 mg/dL |
| Glycated Albumin | 220 mg/dL |

iv. Hemoglobin Variant Interference

A hemoglobin variant study was performed using a total of 121 normal and diabetic whole blood $\mathrm{K}_2\mathrm{EDTA}$ patient variant samples known to contain hemoglobin variants HbS, HbC, HbE, HbD, HbA2, and HbF. Testing was performed using the CAPILLARYS Hb A1c kit on the CAPILLARYS 2 FLEX-PIERCING instrument and results were compared to results obtained by a NGSP reference method that has been demonstrated to be free from the hemoglobin interferent (Trinity Biotech Ultra2 for variants HbS, HbC, HbD, HbE and Tosoh G8 for variants HbA2 and HbF). The sponsor's definition of non-significant interference is $\leq 7\%$ mean relative deviation between the CAPILLARYS Hb A1c result and the NGSP reference method. The range of hemoglobin variants and range of HbA1c that were evaluated are summarized in the table below:

Variant samples used in Hemoglobin Variant Study

| Hemoglobin variant | n | Range in % Hb variant | Range in %HbA1c concentration (NGSP) | Range in mmol/mol HbA1c concentration (IFCC) |
| --- | --- | --- | --- | --- |
| HbS | 20 | 33 – 40.8 | 4.9 – 14 | 30 – 129 |
| HbC | 20 | 28 – 37.2 | 4.6 – 13.1 | 27 – 120 |
| HbD | 21 | 35.5 – 41.3 | 5.0 – 11.8 | 32 – 105 |
| HbE | 20 | 21.3 – 37.0 | 4.8 – 9.8 | 29 – 83 |
| HbA2 | 20 | 4.0 – 7.7 | 7.2 – 11 | 55 – 97 |
| HbF | 20 | 1.5 – 31.4 | 4.9 – 15.1 | 85 – 141 |

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Hemoglobin Variant Study Results

| Hemoglobin Variant | Relative % Bias (range of % bias) to Comparative Method for HbA1c | |
| --- | --- | --- |
| | ~6.5% | ~9.0% |
| HbS | 0.8 % (0.0% – 1.6%) | 0.4% (-3.1% – 3.2%) |
| HbC | 0.0 (0.0% – 0.0%) | 3.3% (1.0% – 5.7%) |
| HbD | 0.5% (-1.4% – 1.6%) | -1.9% (-2.4% – -1.2%) |
| HbE | 0.4% (-3.3% – 1.5%) | -0.3% (-3.7% – 2.1%) |
| HbA2 | -3.4% (-4.5% – -1.4%) | -0.8% (-2.0% – 0.0%) |
| HbF | -3.3% (-4.8% – -1.6%) | -2.0% (-5.0% – 0.0%) |

Significant interference was defined as $\geq 7\%$ change in HbA1c value in the presence of the hemoglobin variant relative to control. The results show there is no significant interference for HbS ($\leq 40.8\%$), HbC ($\leq 37.6\%$), HbD ($\leq 41.3\%$), HbE ($\leq 26.8\%$), and HbA2 ($\leq 7.8\%$) at the concentrations tested in this study.

The software displays the following message if the HbF level is higher than $23\%$: "Atypical profile—possible quantitative interference if HbF or variant > 23%."

The following prominent warning is stated in the package insert:

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

The package insert labeling states:

No interference has been observed with HbA1c fraction quantification due to the presence of major abnormal hemoglobins Hb S ($\leq 40.8\%$), Hb C ($\leq 37.2\%$), Hb D ($\leq 41.3\%$) and Hb E ($\leq 37.0\%$). However, due to the number of variants, the presence of another hemoglobin variant may be observed in the HbA1c migration zone; in the case of a shoulder on HbA1c, no result will be reported by the software (as in presence of Hb Bart's).

f. Assay cut-off:

Not applicable.

2. Comparison studies:

a. Method comparison with predicate device:

A method comparison study of 150 variant-free $\mathrm{K}_2\mathrm{EDTA}$ whole blood samples (4.4 to $16.6\%$ HbA1c) were evaluated using the CAPILLARYS Hb A1c kit on the CAPILLARYS 2 FLEX PIERCING instrument. Results were compared to testing

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performed at a NGSP Secondary Reference Laboratory using a previously cleared HPLC HbA1c assay method (Tosoh HLC-723G8).

The distribution of samples follows in the table below:

| Hemoglobin A1c level | n | % of Samples tested |
| --- | --- | --- |
| ≤ 5% | 7 | 5 |
| 5 – 6% | 19 | 13 |
| 6 – 6.5% | 35 | 23 |
| 6.5 – 7% | 36 | 24 |
| 7 – 8% | 23 | 15 |
| 8 – 9% | 13 | 9 |
| > 9% | 17 | 11 |
| Total samples | 150 | 100.0 |

Linear, Deming (weighted), and Passing-Bablok regression analyses were performed using the CAPILLARYS HbA1c method on the CAPILLARYS 2 FLEX PIERCING instrument versus the comparative G8 HPLC method (NGSP) and are summarized below:

Summary of Method Comparison Results

| | Slope | y-intercept | r |
| --- | --- | --- | --- |
| Linear | 1.027
95% CI (1.019 to 1.035) | -0.265
95% CI (-0.325 to -0.204) | 0.999 |
| Deming | 1.023
95% CI (1.012 to 1.034) | -0.235
95% CI (-0.312 to -0.158) | 0.999 |
| Passing-Bablok | 1.000
95% CI (1.000 to 1.026) | -0.100
95% CI (-0.248 to -0.039) | 0.999 |

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Scatter Plot using Deming Fit, %HbA1c, NGSP vs. CAPILLARYS Hb A1c

Scatter Plot using Passing-Bablok Fit, %HbA1c, NGSP vs. CAPILLARYS Hb A1c

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The following biases between CAPILLARYS Hb A1c run on the CAPILLARYS 2 FLEX-PIERCING instrument versus the NGSP Reference Method (Tosoh G8 HPLC analyzer) were observed:

Bias Estimation

| % HbA1c – Decision Level | Bias | % Bias |
| --- | --- | --- |
| 5.1 | -0.1 | -2.47 |
| 6.4 | -0.1 | -1.42 |
| 8.2 | 0.0 | -0.51 |
| 12.2 | 0.1 | 0.55 |

Total Error Calculations:

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: (5.1%, 6.4%, 8.2% and 12.2%) were calculated as follows: $\% \mathrm{TE} = \% \mathrm{Bias}| + 1.96*\mathrm{CV}^{*}(1 + (\% \mathrm{Bias} / 100))$ . The results are as follows:

Total Error Estimation

| % A1c – Decision Level | % Bias | % CV | % TE |
| --- | --- | --- | --- |
| 5.1 | -2.47 | 1.8 | 5.9 |
| 6.4 | 1.36 | 1.4 | 4.1 |
| 8.2 | 0.50 | 1.2 | 2.8 |
| 12.2 | 0.55 | 1.3 | 3.1 |

b. Matrix comparison:

Matrix comparison studies were previously reviewed in k122101. Only $\mathrm{K}_2\mathrm{EDTA}$ and $\mathrm{K}_3\mathrm{EDTA}$ venous whole blood samples are to be used with this assay.

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

b. Clinical specificity:

Not applicable.

c. Other clinical supportive data (when a. and b. are not applicable):

Not applicable.

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4. Clinical cut-off:

Not applicable.

5. Expected values/Reference range:

| Category | HbA1c Range (NGSP/DCCT) |
| --- | --- |
| Normal | < 5.7% |
| Prediabetes
(increased risk for diabetes) | 5.7% - 6.4% |
| Diabetes | ≥ 6.5% |

The expected HbA1c range for non-diabetic adults is 4 - 6%. However, each laboratory should establish its reference range and HbA1c goal following state and federal regulations and taking into account sex, age, ethnicity and individual patient situation.

The following reference was cited: American Diabetes Association. Standards of medical care in diabetes - 2016. Diabetes Care. 2016 Jan, 39, Suppl 1.

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

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