CAPI 3 Hb A1c
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 |
Indications for 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)
Related Devices
- K162281 — CAPI 3 Hb A1c, MULTI-SYSTEM Hb A1C CAPILLARY CONTROLS (2) · Sebia · Feb 17, 2017
- K171861 — CAPILLARYS Hb A1c · Sebia · Feb 7, 2018
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE
A. 510(k) Number:
k171537
B. Purpose for Submission:
Addition of a diagnostic claim to an existing device
C. Measurand:
Whole blood hemoglobin A1c (HbA1c)
D. Type of Test:
Capillary Electrophoresis
E. Applicant:
Sebia, Inc.
F. Proprietary and Established Names:
CAPI 3 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 Indication for use below.
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2. Indication(s) for 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.
3. Special conditions for use statement(s):
For In Vitro Diagnostic Prescription Use.
The CAPI 3 HbA1c 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 3 TERA instrument
I. Device Description:
The CAPI 3 HbA1c kit is to be used with the CAPILLARYS 3 TERA instrument. The reagents
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are identical to those cleared in k162281. The CAPI 3 HbA1c 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) and filters (4 filters per kit).
CAPILLARYS 3 TERA instrument was cleared in k162281.
## J. Substantial Equivalence Information:
1. Predicate device name(s):
Tosoh Automated Glycohemoglobin Analyzer HLC-723G-8
2. Predicate 510(k) number(s):
k131580
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Candidate Device
Sebia CAPI 3 Hb A1c | Predicate Device
Tosoh HLC-723G8
(k131580) |
| Intended Use | Intended for measurement of % hemoglobin A1c (HbA1c) (DCCT/NGSP) and mmol/mol hemoglobin A1c (IFCC) in whole blood specimens. 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. | Same |
| Standardization | Traceable to the Diabetes Control and Complications Trial (DCCT) reference method and IFCC. Certified via the National Glycohemoglobin Standardization Program | Same |
| Sample type | Venous whole blood | Same |
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| Differences | | |
| --- | --- | --- |
| Item | Candidate Device
Sebia CAPI 3 Hb A1c | Predicate Device
Tosoh HLC-723G8
(k131580) |
| Assay Principle | Capillary electrophoresis | Ion-exchange HPLC |
| Measuring Range | 4.0 – 16.5% (NGSP)
20 – 157 mmol/mol (IFCC) | 4.0 – 16.9% |
## K. Standard/Guidance Document Referenced (if applicable):
CLSI EP05-A3 Evaluation of Precision Performance of Clinical Devices; Approved Guideline
CLSI EP06-A Evaluation of the Linearity of Quantitative Measurement Procedure: A statistical Approach; Approved Guideline
CLSI EP7-A2 Interference Testing in Clinical Chemistry; Approved Guideline-Second Edition
CLSI EP9-A2 Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline
## L. Test Principle:
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 wavelength specific to hemoglobins. Before each run, the capillaries are washed with a wash solution and prepared for the next analysis with buffer.
At the end of analysis, relative quantification of individual HbA1c fractions is performed automatically. The HbA1c concentrations are reported in %HbA1c (DCCT/NGSP) and in mmol/mol (IFCC) units.
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M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
The precision of the CAPI 3 Hb A1c procedure using the CAPILLARYS 3 TERA was evaluated based on CLSI EP05-A3 Evaluation of Precision Performance of Clinical Devices; Approved Guideline. Four K₂ EDTA venous whole blood samples at the targeted HbA1c concentrations of 5.2, 6.5, 8.1, and 11.9 % were used in the study. The study also included two quality control materials (5.2, and 7.7 % HbA1c) and two calibrators (5.5, and 10.1 % HbA1c). The samples were analyzed in duplicate on two capillaries per run, two runs per day using three lots on each of three instruments over 24 days yielding a total of 1728 results over total testing time of 72 days. The results are shown below:
Results in NGSP units (%HbA1c) - % CV by Sample
Instruments Combined
| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Between instrument | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 5.2 | 0.9% | 0.8% | 0.0% | 0.4% | 0.7% | 0.3% | 1.5% |
| 2 Control sample | 5.2 | 0.9% | 0.9% | 0.0% | 0.6% | 0.7% | 0.0% | 1.6% |
| 3 Calibrator sample | 5.5 | 0.8% | 0.8% | 0.0% | 0.5% | 0.7% | 0.0% | 1.4% |
| 4 Human sample | 6.5 | 0.7% | 0.5% | 0.3% | 0.3% | 0.5% | 0.0% | 1.1% |
| 5 Control sample | 7.7 | 0.7% | 0.5% | 0.0% | 0.5% | 0.2% | 1.7% | 2.0% |
| 6 Human sample | 8.1 | 0.7% | 0.5% | 0.0% | 0.4% | 0.3% | 0.1% | 1.0% |
| 7 Calibrator sample | 10.1 | 0.6% | 0.5% | 0.0% | 0.4% | 0.7% | 0.0% | 1.1% |
| 8 Human sample | 11.9 | 0.6% | 0.6% | 0.0% | 0.5% | 1.4% | 0.0% | 1.7% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day, between-lot and between-instrument.
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Instrument 1 (%CV by Sample, NGSP % HbA1c)
| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 5.2 | 0.9% | 0.8% | 0.0% | 0.4% | 0.8% | 0.8% |
| 2 Control sample | 5.2 | 0.8% | 0.9% | 0.0% | 0.6% | 0.9% | 0.9% |
| 3 Calibrator sample | 5.5 | 0.8% | 0.9% | 0.0% | 0.7% | 0.7% | 0.7% |
| 4 Human sample | 6.5 | 0.7% | 0.6% | 0.3% | 0.2% | 0.7% | 0.7% |
| 5 Control sample | 7.7 | 0.6% | 0.6% | 0.0% | 0.4% | 0.1% | 0.1% |
| 6 Human sample | 8.1 | 0.7% | 0.5% | 0.0% | 0.4% | 0.4% | 0.4% |
| 7 Calibrator sample | 10.1 | 0.6% | 0.4% | 0.0% | 0.4% | 1.0% | 1.0% |
| 8 Human sample | 11.9 | 0.6% | 0.6% | 0.0% | 0.5% | 1.8% | 1.8% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
Instrument 2 (%CV by Sample, NGSP % HbA1c)
| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 5.2 | 1.0% | 0.9% | 0.0% | 0.4% | 0.9% | 1.7% |
| 2 Control sample | 5.2 | 0.9% | 1.1% | 0.0% | 0.4% | 0.7% | 1.6% |
| 3 Calibrator sample | 5.5 | 0.8% | 0.8% | 0.0% | 0.4% | 0.8% | 1.4% |
| 4 Human sample | 6.5 | 0.7% | 0.5% | 0.3% | 0.4% | 0.4% | 1.1% |
| 5 Control sample | 7.7 | 0.7% | 0.3% | 0.0% | 0.6% | 0.1% | 1.0% |
| 6 Human sample | 8.1 | 0.8% | 0.3% | 0.1% | 0.3% | 0.1% | 0.9% |
| 7 Calibrator sample | 10.1 | 0.5% | 0.5% | 0.0% | 0.3% | 0.5% | 1.0% |
| 8 Human sample | 11.9 | 0.6% | 0.8% | 0.2% | 0.6% | 1.1% | 1.6% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
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Instrument 3 (%CV by Sample, NGSP % HbA1c)
| Sample | Mean (%) | Within capillary | Between capillary | Between run | Between day | Between lot | Total * |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 5.2 | 0.9% | 0.8% | 0.0% | 0.5% | 0.2% | 1.3% |
| 2 Control sample | 5.2 | 0.9% | 0.8% | 0.0% | 0.6% | 0.6% | 1.5% |
| 3 Calibrator sample | 5.5 | 0.8% | 0.7% | 0.0% | 0.4% | 0.7% | 1.3% |
| 4 Human sample | 6.5 | 0.8% | 0.6% | 0.4% | 0.2% | 0.4% | 1.1% |
| 5 Control sample | 7.7 | 0.6% | 0.6% | 0.0% | 0.4% | 0.3% | 1.0% |
| 6 Human sample | 8.1 | 0.7% | 0.6% | 0.0% | 0.4% | 0.1% | 1.0% |
| 7 Calibrator sample | 10.1 | 0.5% | 0.4% | 0.0% | 0.3% | 0.6% | 1.0% |
| 8 Human sample | 11.9 | 0.5% | 0.6% | 0.0% | 0.4% | 1.1% | 1.4% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
## Results in IFCC units (mmol/mol)
Instruments Combined
| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Between instrument | Total * |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 33 | 1.5% | 1.4% | 0.0% | 0.7% | 1.2% | 0.6% | 2.6% |
| 2 Control sample | 34 | 1.4% | 1.5% | 0.0% | 0.8% | 1.5% | 0.0% | 2.7% |
| 3 Calibrator sample | 37 | 1.2% | 1.4% | 0.0% | 0.7% | 1.1% | 0.0% | 2.3% |
| 4 Human sample | 48 | 1.2% | 0.9% | 0.5% | 0.5% | 0.8% | 0.0% | 1.7% |
| 5 Control sample | 61 | 0.9% | 0.7% | 0.0% | 0.7% | 0.1% | 2.6% | 2.9% |
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| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Between instrument | Total * |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 6 Human sample | 65 | 1.0% | 0.7% | 0.0% | 0.5% | 0.4% | 0.2% | 1.3% |
| 7 Calibrator sample | 86 | 0.7% | 0.6% | 0.0% | 0.5% | 0.8% | 0.0% | 1.3% |
| 8 Human sample | 107 | 0.7% | 0.8% | 0,0% | 0.6% | 1.7% | 0.0% | 2.1% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day, between-lot and between-instrument.
Instrument 1 (%CV by Sample, IFCC units)
| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 33 | 1.5% | 1.3% | 0.0% | 0.6% | 1.3% | 2.4% |
| 2 Control sample | 34 | 1.3% | 1.4% | 0.0% | 0.8% | 1.8% | 2.8% |
| 3 Calibrator sample | 37 | 1.3% | 1.2% | 0.0% | 0.8% | 1.2% | 2.3% |
| 4 Human sample | 48 | 1.1% | 0.8% | 0.4% | 0.6% | 1.0% | 1.9% |
| 5 Control sample | 61 | 0.9% | 0.8% | 0.0% | 0.6% | 0.1% | 1.4% |
| 6 Human sample | 65 | 0.9% | 0.6% | 0.0% | 0.5% | 0.8% | 1.3% |
| 7 Calibrator sample | 86 | 0.8% | 0.5% | 0.0% | 0.5% | 1.2% | 1.6% |
| 8 Human sample | 107 | 0.6% | 0.7% | 0.0% | 0.6% | 2.2% | 2.5% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
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Instrument 2 (%CV by Sample, IFCC units)
| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 33 | 1.7% | 1.7% | 0.0% | 0.7% | 1.4% | 2.8% |
| 2 Control sample | 34 | 1.6% | 1.7% | 0.0% | 0.7% | 1.3% | 2.7% |
| 3 Calibrator sample | 37 | 1.2% | 1.5% | 0.0% | 0.6% | 0.8% | 2.2% |
| 4 Human sample | 48 | 1.3% | 0.9% | 0.8% | 0.3% | 1.6% | 1.9% |
| 5 Control sample | 61 | 0.9% | 0.5% | 0.0% | 0.8% | 0.0% | 1.3% |
| 6 Human sample | 65 | 1.1% | 0.5% | 0.0% | 0.5% | 0.0% | 1.3% |
| 7 Calibrator sample | 86 | 0.7% | 0.6% | 0.0% | 0.5% | 0.5% | 1.1% |
| 8 Human sample | 107 | 0.8% | 0.9% | 0.0% | 0.7% | 1.3% | 1.9% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
Instrument 3 (%CV by Sample, IFCC units)
| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 1 Human sample | 33 | 1.3% | 1.4% | 0.0% | 0.9% | 1.0% | 2.3% |
| 2 Control sample | 34 | 1.4% | 1.5% | 0.0% | 0.9% | 1.2% | 2.6% |
| 3 Calibrator sample | 37 | 1.1% | 1.5% | 0.0% | 0.8% | 1.3% | 2.4% |
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| Sample | Mean (mmol/mol) | Within capillary | Between capillary | Between run | Between day | Between lot | Total* |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 4 Human sample | 48 | 1.1% | 1.0% | 0.4% | 0.4% | 0.7% | 1.7% |
| 5 Control sample | 61 | 0.9% | 0.7% | 0.0% | 0.7% | 0.2% | 1.3% |
| 6 Human sample | 65 | 0.9% | 0.8% | 0.0% | 0.4% | 0.2% | 1.3% |
| 7 Calibrator sample | 86 | 0.7% | 0.5% | 0.0% | 0.4% | 0.7% | 1.2% |
| 8 Human sample | 107 | 0.6% | 0.6% | 0.0% | 0.5% | 1.3% | 1.7% |
*Total Reproducibility includes: within-capillary, between-capillary, between-run, between-day and between-lot.
# b. Linearity/assay reportable range:
The linearity of the CAPI 3 HbA1c procedure was evaluated based on CLSI EP06-A Guideline Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach. Two blood samples, including a normal sample with HbA1c concentration at $3.8\%$ HbA1c (18 mmol/mol) and an elevated HbA1c level sample with HbA1c concentration at $17.3\%$ HbA1c (166 mmol/mol) 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 following table summarizes the linear regression results.
| Units | Slope | y-intercept | Correlation coefficient |
| --- | --- | --- | --- |
| NGSP (%HbA1c) | 1.0034 | -0.3047 | 0.999 |
| IFCC (mmol/mol) | 1.0016 | -3.2377 | 0.999 |
The study supports the claimed reportable range of the device from 4.0 to $16.5\%$ HbA1c (20 - 157 mmol/mol).
# c. Traceability, Stability, Expected values (controls, calibrators, or methods):
# Traceability:
The CAPI 3 HbA1c assay 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.
# Value assignment:
The Hb A1c CAPILLARY CALIBRATORS are required for use with this device. Value
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assignment, stability protocols and acceptance criteria were previously reviewed in k162281 and k122101.
# Stability:
The stability protocols and acceptance criteria for the different components of the CAPI 3 HbA1c kit were reviewed and determined to be adequate in k162281.
The expiration dates of the different components of the CAPI 3 HbA1c kit are indicated as follows:
| Kit Component | Shelf-Life | On Board Stability |
| --- | --- | --- |
| CAPI 3 HbA1c hemolysing solution | 3 years at 2-8°C and 15-30°C | 2 months |
| CAPI 3 HbA1c buffer | 3 years at 2-8°C | 1 month |
| CAPI 3 HbA1c wash solution | 3 years at 2-8°C and 15-30°C | 121 days |
# d. Detection limit:
The detection limit studies were previously reviewed and cleared in k162281. Those studies were not repeated because the device and procedure has not changed; the intended use has been changed to include a diagnostic claim.
# e. Analytical specificity:
# i. Endogenous Interference
Interference studies were performed to assess endogenous substances that could interfere with the CAPI 3 HbA1c assay. The interfering substances were evaluated in venous whole blood $\mathrm{K}_2$ EDTA samples that contained a blood sample close to the cut-off value $6.4\%$ (46 mmol/mol) and a blood sample with elevated HbA1c level $8.4\%$ (69 mmol/mol). Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor's definition of non-significant interference is $\leq 7\%$ mean relative deviation between the tested and the control samples.
Concentration at which no significant interference $(\leq 7\%)$ was observed
| Endogenous interfering factor | Concentration |
| --- | --- |
| Bilirubin | 70 mg/dL (1197 μM) |
| D-glucose | 1000 mg/dL (55 mM) |
| 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) |
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ii. Exogenous Interference
Interference studies were performed to assess common or known exogenous substances that could interfere with the CAPI 3 HbA1c assay. The interfering substances were evaluated in venous whole blood K₂EDTA samples that contained a blood sample close to the cut-off value 6.4% (46 mmol/mol) and a blood sample with elevated HbA1c level 8.4% (69 mmol/mol). Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor’s definition of non-significant interference is ≤ 7% mean relative deviation between the tested and the control samples
Concentration at which no significant interference (≤ 7%) was observed
| Potential interfering substance | Concentration |
| --- | --- |
| Acetaminophen | 200 mg/L (1325 μM) |
| Acetylcysteine | 200 mg/dL (12.3 mM) |
| Acetylsalicylic 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 |
| 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) |
iii. Cross Reactivity with Hemoglobin Derivatives
To study interference from carbamylated hemoglobin, four EDTA whole blood patient samples with HbA1c concentrations at 6.3% HbA1c, 8.3% HbA1c, 45 mmol/mol HbA1c and 67 mmol/mol HbA1c were split into two aliquots. One aliquot, at each HbA1c level, was spiked with 1 mmol/L of potassium cyanate and incubated for 3 hours and 30 minutes at 37°C. Another aliquot, at each HbA1c level, without potassium cyanate was incubated for 3 hours and 30 minutes at 37°C. Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor’s definition of non-significant interference is ≤7% mean relative deviation between the tested and the control samples. The sponsor concluded that presence of ≤ 8.1 mg/mL carbamylated hemoglobin does not interfere with this assay.
To study interference from labile hemoglobin, four EDTA whole blood patient samples
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with HbA1c concentrations at $6.3\%$ HbA1c, $8.5\%$ HbA1c, 45 mmol/mol HbA1c and 67 mmol/mol HbA1c were split into two aliquots. One aliquot, at each HbA1c level, was spiked with glucose $(50~\mathrm{g / L})$ and incubated for 3 hours at $37^{\circ}\mathrm{C}$ . Another aliquot, at each HbA1c level, without glucose was incubated for 3 hours at $37^{\circ}\mathrm{C}$ . Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor's definition of non-significant interference is $\leq 7\%$ mean relative deviation between the tested and the control samples. The sponsor concluded that labile HbA1c $\leq$ $20.0~\mathrm{mg / mL}$ does not interfere with this assay.
To study interference from acetylated hemoglobin, four EDTA whole blood samples with HbA1c concentrations at $6.2\%$ HbA1c, $8.4\%$ HbA1c, 44 mmol/mol HbA1c and 68 mmol/mol HbA1c were split into two aliquots. One aliquot, at each HbA1c level, was spiked with acetylsalicylic acid (10 mmol/L) and incubated for 6 hours at $37^{\circ}\mathrm{C}$ . Another aliquot, at each HbA1c level, without acetylsalicylic acid was incubated for 6 hours at $37^{\circ}\mathrm{C}$ . Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor's definition of non-significant interference is $\leq 7\%$ mean relative deviation between the tested and the control samples. The sponsor concluded that acetylated hemoglobin up to $\leq 4.2\mathrm{mg / mL}$ does not interfere with this assay.
To study interference from glycated albumin, four EDTA whole blood samples with HbA1c concentrations at $6.2\%$ HbA1c, $8.5\%$ HbA1c, 44 mmol/mol HbA1c and 70 mmol/mol HbA1c were split into two aliquots. One aliquot, at each HbA1c level, was spiked with glycated albumin $(0.20\mathrm{mg / mL})$ and the other aliquot, at each HbA1c level, was without glycated albumin. Ten replicates were analyzed using the CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system. The sponsor's definition of non-significant interference is $\leq 7\%$ mean relative deviation between the tested and the control samples. The sponsor concluded that glycated albumin up to $\leq 0.20\mathrm{mg / mL}$ does not interfere with this assay.
# iv. Hemoglobin Variant Interference
A hemoglobin variant study was performed using a total of 131 normal and diabetic whole blood $\mathrm{K}_2\mathrm{EDTA}$ 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, and D, A2, and F were performed using CAPI 3 Hb A1c on the CAPILLARYS 3 TERA testing system and compared to results obtained by a NGSP reference method that has been demonstrated to be free from interference by the hemoglobin variants (Trinity Biotech Ultra2 for HbS, HbC, HbD, HbE and Tosoh HLC-723G8 for HbF and HbA2).
Variant samples used in Hemoglobin Variant Study
| Hemoglobin Variant | n | Range in % HbVariant | Range in %HbA1c Concentration |
| --- | --- | --- | --- |
| Hb A2 | 20 | 4.0-7.8 | 4.5-11.2 |
| Hb F | 19 | 1.5-23.7 | 5.1-15.1 |
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| 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 |
Hemoglobin Variant Study Bias Results
| Hemoglobin Variant | Relative % Bias to Comparative Method | |
| --- | --- | --- |
| | Relative %Bias (Range of %Bias) for HbA1c | Relative %Bias (Range of %Bias) for HbA1c |
| | ~6.5% | ~9.0% |
| HbS | 1.6% (0% - 3.2%) | 2.9% (1.1% - 5.4%) |
| HbC | -1.8% (-3.1% - 0%) | 3.9% (3.3% - 4.5%) |
| HbD | 1.0% (0% - 1.6%) | 0.8% (0% - 2.4%) |
| HbE | 1.5% (-1.5% - 1.6%) | 1.2% (0%-2.5%) |
| HbA2 | 0.7% (0% -1.5%) | 0.4% (0% - 1.1%) |
| HbF | -4.1% (-3.3% - 4.9%) | -0.8% (0% - 2.0%) |
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 ( $\leq 7.8\%$ ) and HbF ( $\leq 23.7\%$ ) at the concentrations tested in this study.
The package insert labeling contains the following prominent boxed warning;
"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 ( $\leq 37.6\%$ ), Hb D ( $\leq 41.3\%$ ) and Hb E ( $\leq 26.8\%$ ). 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.
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# 2. Comparison studies:
# a. Method comparison with predicate device:
A method comparison study of 152 variant-free $\mathrm{K}_2\mathrm{EDTA}$ whole blood samples (3.9 - $16.5\%$ HbA1c concentration range) were evaluated using the CAPI 3 Hb A1c kit and the CAPILLARYS 3 TERA instrument. The results were compared to the testing performed at an NGSP secondary reference laboratory using a previously cleared HPLC HbA1c method (Tosoh HLC-723G8). The distribution of samples follows in the table below.
Distribution of samples
| Hemoglobin A1c level | n | % of Samples tested |
| --- | --- | --- |
| ≤ 5% | 9 | 6 |
| 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 | 152 | 100.0 |
Linear, Deming (weighted), and Passing-Bablok regression analyses were performed for the CAPI 3 Hb A1c kit and the CAPILLARYS 3 TERA instrument versus the comparative Tosoh HLC-723G8 method (NGSP) and are summarized below:
Summary of Method Comparison Results
| | Slope | 95% CI | y-Intercept | 95% CI | R² |
| --- | --- | --- | --- | --- | --- |
| Linear | 1.014 | 1.007 to 1.021 | -0.142 | -0.197 to -0.087 | 0.999 |
| Deming | 1.011 | 1.001 to 1.021 | -0.123 | -0.196 to -0.050 | 0.999 |
| Passing-Bablok | 1.000 | 1.000 to 1.012 | 0.000 | -0.115 to 0.000 | 0.999 |
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Scatter Plot using Linear Regression Fit, %HbA1c, NGSP vs. CAPI 3 Hb A1c
Scatter Plot using Deming Fit, %HbA1c, NGSP vs. CAPI 3 Hb A1c
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Scatter Plot using Passing-Bablok Fit,
\% \mathrm{HbA1c}
, NGSP vs. CAPI 3 Hb A1c
# 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: $(5.0\%, 6.5\%, 8.0\%$ and $12.0\%)$ 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 |
| --- | --- | --- | --- |
| 5.2 | -1.32 | 1.6 | 4.4 |
| 6.5 | -0.77 | 1.1 | 2.9 |
| 8.1 | -0.34 | 1.0 | 2.3 |
| 11.9 | 0.22 | 1.7 | 3.6 |
# b. Matrix comparison:
The matrix comparison studies were previously reviewed in k162281. Only $\mathrm{K}_2$ EDTA and $\mathrm{K}_3$ EDTA venous whole blood samples are to be used with this assay.
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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.
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 in their country of business 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.
