D-100 HbA1c, D-100 HbA1c Calibrator Pack
Device Facts
| Record ID | K151321 |
|---|---|
| Device Name | D-100 HbA1c, D-100 HbA1c Calibrator Pack |
| Applicant | Bio-Rad Laboratories, Inc. |
| Product Code | PDJ · Clinical Chemistry |
| Decision Date | Dec 9, 2015 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1373 |
| Device Class | Class 2 |
Indications for Use
The D-100™ HbA1c test 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-100 Hemoglobin Testing System. Hemoglobin A1c measurements are used as an aid in diagnosis of diabetes mellitus, 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-100TM HbA1c test is intended for Professional Use Only. Calibrators: The D-100™ HbA1c Calibrator Pack is for the calibration of the D-100 Hemoglobin Testing System used for the quantitative determination of hemoglobin A1c(HbA1c) in human whole blood.
Device Story
The D-100™ HbA1c is an in vitro diagnostic test for professional use on the D-100 Hemoglobin Testing System. It utilizes ion-exchange high-performance liquid chromatography (HPLC) to quantify hemoglobin A1c in human whole blood. The system automatically hemolyzes and dilutes samples before introducing them into an analytical cartridge. A programmed buffer gradient separates hemoglobin species based on ionic interactions. A detector measures absorbance at 415 nm, producing a digital chromatogram. Software analyzes peaks (retention time, area, relative percent) to calculate HbA1c levels. An optional 'Advisor' feature compares reports against user-defined rules. Results are reported in NGSP (%) and IFCC (mmol/mol) units. The device aids clinicians in diagnosing diabetes, identifying at-risk patients, and monitoring long-term glycemic control.
Clinical Evidence
No clinical trials; performance established via bench testing. Method comparison study (n=129) against NGSP Secondary Reference Laboratory (Tosoh G8) showed strong correlation (Deming slope 0.9867, Passing-Bablok slope 0.9909). Precision evaluated per CLSI EP05-A2 (Total CV 0.9-2.0%). Linearity confirmed 3.5-20% HbA1c range. Interference studies (CLSI EP07-A2) confirmed no significant interference from endogenous substances, drugs, or common hemoglobin variants (S, C, D, E, F, A2).
Technological Characteristics
Ion-exchange HPLC; cation exchange cartridge. Reagents: succinate/sodium perchlorate buffers, deionized water, sodium azide preservative. Lyophilized human whole blood calibrators (glycine/trehalose). Automated hemolysis/dilution. Connectivity: Barcode/manual entry, no wireless transmission. Software-controlled peak identification and analysis. Sterilization: Not applicable (reagents).
Indications for Use
Indicated for quantitative determination of HbA1c in human whole blood for diabetes diagnosis, risk identification, and long-term glucose monitoring. Not for newborns, pediatric patients, pregnant women, or Type 1 diabetes glucose monitoring. Not for gestational diabetes diagnosis. Contraindicated in patients with conditions altering red blood cell lifespan (e.g., hemolytic anemia, hemoglobinopathies, thalassemias, recent blood loss/transfusion, severe hepatic/renal disease, malignancies).
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
Related Devices
- K161687 — D-10 Hemoglobin A1c Program · Bio-Rad Laboratories, Inc. · Oct 14, 2016
- K162822 — ADAMS A1c HA-8180V, CALIBRATOR 80 · Arkray Factory, Inc. · Jun 29, 2017
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY AND INSTRUMENT COMBINATION TEMPLATE
A. 510(k) Number:
K151321
B. Purpose for Submission:
New Device
C. Measurand:
Whole blood Glycosylated Hemoglobin (HbA1c)
D. Type of Test:
Ion-Exchange Quantitative High Performance Liquid Chromatography
E. Applicant:
Bio-Rad Laboratories, Inc.
F. Proprietary and Established Names:
D-100 HbA1c
D-100 HbA1c Calibrator Pack
G. Regulatory Information:
| Regulatory Description | Product Code | Classification | Regulation Section | Panel |
| --- | --- | --- | --- | --- |
| Hemoglobin A1c Test System | PDJ, LCP | Class II | 21 CFR 862.1373 | Chemistry, 75 |
| Calibrator | JIT | Class II | 21 CFR 862.1150 | Chemistry, 75 |
H. Intended Use:
1. Intended use(s):
See indications for use below.
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2. Indication(s) for use:
The D-100 HbA1c test 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-100 Hemoglobin Testing System.
Hemoglobin A1c measurements are used as an aid in diagnosis of diabetes, as an aid in identifying patients who may be at risk for developing diabetes mellitus, and for the monitoring of long-term blood glucose control individuals with diabetes mellitus.
The Bio-Rad D-100 HbA1c test is intended for Professional Use Only.
Calibrators:
The D-100 HbA1c Calibrator Pack is for the calibration of the D-100 Hemoglobin Testing System used for the quantitative determination of hemoglobin A1c (HbA1c) in human whole blood.
3. Special conditions for use statement(s):
The D-100 HbA1c test is not intended for analysis of samples collected from newborns.
The D-100 HbA1c test should not be used to replace glucose testing in pediatric patients, pregnant women, or patients with Type 1 diabetes.
HbA1c should not be used 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.
The D-100 HbA1c test should not be used 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.
For prescription use only.
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4. Special instrument requirements:
Bio-Rad D-100 Hemoglobin Testing System
I. Device Description:
The D-100 HbA1c assay is run on the D-100 Hemoglobin Testing System instrument.
The Bio-Rad D-100 HbA1c assay includes the following components:
100 TM Prefilters
2000 tests each. Package of 5
100 TM Cleaning Tube.
One microvial containing 1.5 mL of a liquid cleaning solution.
100 TM Sample Diluent.
Each bottle contains 1 L of deionized water with <0.1% sodium azide as a preservative.
100 TM HbA1c Elution Buffer A.
Each bottle contains 2600 mL of a succinate/sodium perchlorate buffer. Contains <0.1% sodium azide as a preservative.
100 TM HbA1c Elution Buffer B.
Each bottle contains 1400 mL of a succinate/sodium perchlorate buffer. Contains <0.1% sodium azide as a preservative.
100 TM HbA1c Wash Solution.
Each bottle contains 3300 mL of deionized water with <0.1% sodium azide as a preservative.
D-100 TM HbA1c Analytical Cartridge/Calibrator Pack. One pack consisting of:
- Cation exchange cartridge. 10,000 tests each
- Calibrator Pack: 1 vial of Conditioner, 1 vial of Calibrator Level 1, and 1 vial of Calibrator Level 2. The vials contain lyophilized human whole blood with glycine and trehalose as preservatives.
- Each Calibrator Pack contains Calibrator values which have been value assigned using secondary calibrators that are traceable to the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference method.
The Calibrators contain lyophilized human whole blood with glycine and trehalose as preservatives.
Each unit of whole blood used in the manufacture of the calibrators and conditioner was tested by FDA accepted methods and found non-reactive for HIV-1, HIV-2, Hepatitis B (HBV), Hepatitis C (HCV), and syphilis.
Controls for the system were previously reviewed and cleared in k070546 and k052838.
J. Substantial Equivalence Information:
1. Predicate Device name(s):
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VARIANT II TURBO HbA1c Kit -2.0
VARIANT II Hemoglobin A1c Calibrators
2. Predicate 510(k) number(s)
K142448
K070452
3. Comparison with predicate
| Similarities and Differences - Assay | | |
| --- | --- | --- |
| Item | Candidate Device
D-100HbA1c
(K151321) | Predicate Device:
VARIANT II TURBO
HbA1c Kit - 2.0
(K142448) |
| Intended Use | Intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes, and for the monitoring of long-term blood glucose control in individuals with diabetes mellitus. | Intended for the quantitative determination of hemoglobin A1c (IFCC mmol/mol and NGSP %) as an aid in diagnosis of diabetes and as an aid in identifying patients who may be at risk for developing diabetes. |
| Test principle | Ion exchange HPLC | Same |
| Sample Types | Human Whole Blood collected with K2-EDTA, K3-EDTA
Potassium Oxalate/Sodium Fluoride, Sodium Citrate, Sodium Heparin, Lithium Heparin | Human Whole Blood collected with K2-EDTA, K3-EDTA
Hemoglobin Capillary Collection Kit |
| Instrument Platform | D-100™ Hemoglobin Testing System | VARIANT™TURBO Hemoglobin Testing System and VARIANT™TURBO Link Hemoglobin Testing System |
| Measuring Range | 3.5 to 20% (NSGP)
15 – 195 mmol/mol
HbA1c (IFCC) | 3.4 to 20.6 % (NSGP)
14 – 203 mmol/mol
HbA1c (IFCC) |
| Standardization | Traceable to the Diabetes Control and Complications Trial (DCCT) reference method and | Same |
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| Similarities and Differences - Assay | | |
| --- | --- | --- |
| Item | Candidate Device D-100HbA1c (K151321) | Predicate Device: VARIANT II TURBO HbA1c Kit – 2.0 (K142448) |
| | IFCC. Certified via the National Glycohemoglobin Standardization Program (NGSP). | |
| Similarities and Differences - Calibrators | | |
| --- | --- | --- |
| Item | Candidate Device D-100 HbA1c Calibrator Pack (K151321) | Predicate Device: VARIANT II Hemoglobin A1c Calibrators (K070452) |
| Intended Use | Intended for the quantitative determination of hemoglobin A1c in Human Whole Blood | Same |
| Levels | Levels 1 & 2 Calibration is performed once at the beginning of a new cartridge. | Same |
| Standardization/Traceability | Each lot of calibrators is value assigned and values are reported in both NGSP and IFCC units. | Same |
| Matrix | Lyophilized human whole blood with glycine and trehalose as preservatives. | Human red blood cell hemolysate with gentamicin, tobramycin, and EDTA as preservatives. |
# K. Standard/Guidance Document Referenced (if applicable):
CLSI EP05-A2: Evaluation of Precision Performance of Quantitative Measurements Methods; Approved Guidelines - Second Edition
CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedure: A Statistical Approach: Approved Guideline
CLSI EP07-A2: Interference Testing in Clinical Chemistry: Approved Guideline- Second Edition
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CLSI EP09-A2-IR: Method Comparison and Bias Estimation Using Patient samples; Approved Guideline- Second Edition
CLSI EP14-A2: Evaluation of Matrix Effect: Approved Guideline-Second Edition
# L. Test Principle:
The device is based on chromatographic separation of HbA1c on a cation exchange cartridge. A high pressure pumping system first delivers a buffer solution to a cation exchange cartridge and detector. Whole blood samples undergo automatic hemolysis and dilution before being delivered to the cation exchange cartridge in a buffer gradient of increasing ionic strength. The various forms of hemoglobin exhibit charge differences at the pH of the buffer and are separated on the cation exchange cartridge based on these charge differences. The separated hemoglobin species then pass through a flow cell where changes in absorbance are measured and recorded as a digital chromatogram. Software performs an analysis of the hemoglobin peaks in the chromatogram and identifies peaks as target analytes. The software includes an optional feature (Advisor) that compares the sample report against a set of rules that are programmed to take user-specified actions.
# M. Performance Characteristics (if/when applicable):
# 1. Analytical performance:
# a. Precision/Reproducibility:
Precision of the D-100 HbA1c test was evaluated based on CLSI EP05-A2 guidelines, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guidelines - Second Edition. Four $\mathrm{K}_3$ -EDTA whole blood samples at concentrations near $5\%$ , $6.5\%$ , $8\%$ and $12\%$ HbA1c and five whole blood quality control solutions (Control 1, Control 2, QC1, QC2, QC 3) were analyzed in duplicate, twice a day, with three lots of reagents, over 20 days each, on three D-100 Hemoglobin Testing System instruments at two different sites. For each sample there were 720 measurements. Results are shown in the tables below.
Table 1: Instrument 1 (%CV by Sample, NGSP)
| Variation Source | Instrument ID: (SM93) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (NGSP%) | 5.5 | 9.4 | 5.2 | 6.7 | 8.1 | 12.0 | 5.3 | 9.9 | 14.8 |
| Repeatability | 0.7 | 0.7 | 0.7 | 0.7 | 0.7 | 0.6 | 0.8 | 0.7 | 0.7 |
| Between-Run | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 0.6 | 0.2 | 0.3 | 0.2 | 0.1 | 0.3 | 0.3 | 0.1 | 0.2 |
| Between-Lot | 1.2 | 0.8 | 1.3 | 1.1 | 1.0 | 0.6 | 1.4 | 0.8 | 0.6 |
| Total Precision | 1.5 | 1.1 | 1.5 | 1.3 | 1.2 | 0.9 | 1.6 | 1.0 | 0.9 |
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Table 2: Instrument 2 (%CV by Sample, NGSP)
| Variation Source | Instrument ID: (SM94) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (NGSP%) | 5.5 | 9.4 | 5.2 | 6.6 | 8.1 | 12.0 | 5.3 | 10.0 | 14.8 |
| Repeatability | 1.1 | 0.9 | 0.8 | 0.9 | 1.0 | 0.9 | 1.0 | 1.0 | 1.0 |
| Between-Run | 0.0 | 0.3 | 0.0 | 0.0 | 0.2 | 0.0 | 0.0 | 0.1 | 0.1 |
| Between-Day | 0.6 | 0.2 | 0.5 | 0.5 | 0.3 | 0.3 | 0.2 | 0.4 | 0.3 |
| Between-Lot | 1.2 | 0.2 | 1.5 | 0.6 | 0.0 | 0.3 | 1.5 | 0.1 | 0.5 |
| Total Precision | 1.7 | 1.0 | 1.8 | 1.2 | 1.0 | 1.0 | 1.8 | 1.1 | 1.1 |
Table 3: Instrument 3 (%CV by Sample, NGSP)
| Variation Source | Instrument ID: (SM98) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (NGSP%) | 5.4 | 9.4 | 5.1 | 6.6 | 8.1 | 12.0 | 5.3 | 9.9 | 14.7 |
| Repeatability | 1.0 | 1.0 | 1.0 | 1.1 | 0.9 | 0.9 | 1.0 | 0.9 | 0.8 |
| Between-Run | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 0.6 | 0.5 | 0.4 | 0.5 | 0.5 | 0.4 | 0.4 | 0.4 | 0.4 |
| Between-Lot | 1.6 | 1.0 | 1.5 | 1.5 | 1.2 | 1.1 | 1.7 | 1.0 | 1.1 |
| Total Precision | 1.9 | 1.4 | 1.9 | 2.0 | 1.6 | 1.5 | 2.0 | 1.4 | 1.4 |
Table 4: Instruments Combined (%CV by Sample, NGSP)
| Variation Source | All Instruments | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (NGSP %) | 5.5 | 9.4 | 5.2 | 6.6 | 8.1 | 12.0 | 5.3 | 9.9 | 14.8 |
| Repeatability | 0.9 | 0.9 | 0.9 | 0.9 | 0.9 | 0.8 | 0.9 | 0.9 | 0.8 |
| Between-Run | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 0.6 | 0.3 | 0.4 | 0.4 | 0.4 | 0.3 | 0.3 | 0.3 | 0.3 |
| Between-Instrument | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Lot | 1.4 | 0.7 | 1.5 | 1.1 | 0.9 | 0.7 | 1.5 | 0.7 | 0.8 |
| Total Precision | 1.7 | 1.2 | 1.7 | 1.5 | 1.3 | 1.2 | 1.8 | 1.2 | 1.2 |
Table 5: Instrument 1 (%CV by Sample, IFCC)
| Variation Source | Instrument ID: (SM93) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (IFCC mmol/mol) | 36.5 | 79.3 | 33.2 | 49.4 | 65.4 | 108.1 | 34.5 | 85.0 | 137.7 |
| Repeatability | 1.2 | 0.9 | 1.2 | 1.0 | 0.9 | 0.8 | 1.3 | 0.8 | 0.8 |
| Between-Run | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 0.9 | 0.3 | 0.5 | 0.4 | 0.2 | 0.4 | 0.6 | 0.2 | 0.3 |
| Between-Lot | 2.0 | 1.1 | 2.2 | 1.6 | 1.3 | 0.7 | 2.3 | 1.0 | 0.7 |
| Total Precision | 2.5 | 1.4 | 2.5 | 1.9 | 1.6 | 1.1 | 2.7 | 1.3 | 1.1 |
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Table 6: Instrument 2 (%CV by Sample, IFCC)
| Variation Source | Instrument ID: (SM94) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (IFCC mmol/mol) | 36.4 | 79.2 | 32.9 | 49.0 | 65.1 | 108.0 | 34.5 | 85.4 | 137.9 |
| Repeatability | 1.7 | 1.2 | 1.4 | 1.4 | 1.3 | 1.1 | 1.7 | 1.3 | 1.1 |
| Between-Run | 0.0 | 0.4 | 0.0 | 0.0 | 0.3 | 0.0 | 0.0 | 0.1 | 0.1 |
| Between-Day | 1.0 | 0.3 | 0.8 | 0.7 | 0.4 | 0.4 | 0.3 | 0.5 | 0.4 |
| Between-Lot | 2.1 | 0.3 | 2.6 | 1.0 | 0.0 | 0.4 | 2.5 | 0.1 | 0.6 |
| Total Precision | 2.9 | 1.4 | 3.1 | 1.8 | 1.4 | 1.2 | 3.1 | 1.4 | 1.3 |
Table 7: Instrument 3 (%CV by Sample, IFCC)
| Variation Source | Instrument ID: (SM98) | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (IFCC mmol/mol) | 36.0 | 79.1 | 32.7 | 48.9 | 64.9 | 107.7 | 34.1 | 84.8 | 137.6 |
| Repeatability | 1.6 | 1.2 | 1.8 | 1.7 | 1.3 | 1.1 | 1.6 | 1.2 | 1.0 |
| Between-Run | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 1.0 | 0.6 | 0.8 | 0.8 | 0.7 | 0.5 | 0.7 | 0.5 | 0.5 |
| Between-Lot | 2.6 | 1.2 | 2.6 | 2.3 | 1.6 | 1.3 | 2.9 | 1.2 | 1.2 |
| Total Precision | 3.2 | 1.8 | 3.3 | 2.9 | 2.1 | 1.8 | 3.4 | 1.7 | 1.7 |
Table 8: Instruments Combined (%CV by Sample, IFCC)
| Variation Source | All Instruments | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Control 1 | Control 2 | Patient 1 | Patient 2 | Patient 3 | Patient 4 | QC 1 | QC 2 | QC 3 |
| Concentration HbA1c (IFCC mmol/mol) | 36.3 | 79.2 | 33.0 | 49.1 | 65.1 | 107.9 | 34.3 | 85.1 | 137.8 |
| Repeatability | 1.5 | 1.1 | 1.5 | 1.4 | 1.2 | 1.0 | 1.5 | 1.1 | 1.0 |
| Between-Run | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Day | 1.0 | 0.4 | 0.7 | 0.6 | 0.5 | 0.4 | 0.6 | 0.4 | 0.4 |
| Between-Instrument | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| Between-Lot | 2.2 | 1.0 | 2.5 | 1.7 | 1.2 | 0.9 | 2.6 | 0.9 | 0.9 |
| Total Precision | 2.9 | 1.5 | 3.0 | 2.3 | 1.7 | 1.4 | 3.1 | 1.5 | 1.4 |
b. Linearity/assay reportable range:
A linearity study was performed based on CLSI EP06-A: Evaluation of the Linearity of Quantitative Measuring Procedures; Linearity across the reportable range was performed using altered patient samples collected using $\mathrm{K}_3$-EDTA. Samples were prepared by first altering two patient whole blood samples to obtain a low HbA1c of $3.5\%$ (15 mmol/mol) and a high HbA1c of $20.0\%$ (195 mmol/mol). These high and low samples were mixed together in varying ratios to obtain 9 additional intermediate
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sample levels. The eleven test samples (high, low and 9 intermediates) were run in duplicate on one instrument; measured values were compared to expected values.
The regression parameters (slope, intercept, and R²) were the following:
Table 9: linearity Regression parameters
| Units/Values | Slope | Intercept | R² |
| --- | --- | --- | --- |
| NGPSP | 0.9992 | 0.03961 | 0.9997 |
| IFCC | 0.999 | 0.455 | 0/9997 |
The linearity study was reviewed and found acceptable. Results of the linearity study support the claimed measuring range of the device of 3.5% to 20% HbA1c (15-195 mmol/mol).
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability
D-100 HbA1c test standardization is traceable to both the International Federation of Clinical Chemistry (IFCC) reference calibrators and the Diabetes Control and Complications Trial (DCCT). The assay is certified by the National Glycohemoglobin Standardization Program (NGSP). The NGSP certification expires in one year. See the NGSP website for current certification at http://www.ngsp.org.
HbA1c results are provided to users in two different units: NGSP equivalent units (%) and IFCC equivalent units (mmol/mol). Results in % HbA1c from the NGSP correlation are calculated from the individual quantitative results for Hemoglobin A1c. The IFCC units of mmol/mol are calculated using the Master Equation: NGSP (%) = 0.09148 x IFCC (mmol/mol) + 2.152.
Closed-Vial Stability:
Accelerated and real time stability study protocols and acceptance criteria for reagents including the Analytical Cartridge, Prefilter, Sample Diluent, Elution Buffer A, Elution Buffer B, Wash Solution and Cleaning Tube were reviewed and found to be acceptable. The Analytical Cartridge and Prefilter are stable for 24 months when stored at 2–8°C, the Sample Diluent is stable for 24 months when stored unopened at 15–35°C, Elution Buffers and Wash Solution are stable for 24 months when stored unopened at 2–8°C, the Cleaning Tube is stable for 12 months when stored unopened at 15–35°C. Real time stability studies are ongoing.
In-Use Stability:
In-use stability study protocols and acceptance criteria for reagents including the Analytical Cartridge, Prefilter, Sample Diluent, Elution Buffer A, Elution Buffer B, and Wash Solution were reviewed and found to be acceptable. The Analytical
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Cartridge and Prefilter are stable for 90 days when stored at 15–35°C when installed on the instrument, the Sample Diluent is stable for 90 days after opening when stored at 15–35°C, the Elution Buffers and Wash Solution are stable for 90 days at 15–35°C when installed on the instrument.
## Calibrators and Control Materials
### Value assignment:
Value assignment for the D-100 HbA1c Calibrator Pack (Level 1 and Level 2) is conducted by measuring each Level over multiple runs using multiple D-100 Hemoglobin Testing System instruments. The D-100 HbA1c Calibrator Pack Levels are assigned final concentration values based on the mean of multiple runs and comparison to values of IFCC reference calibrators run in parallel.
The commercially available Bio-Rad Lyphocheck Diabetes control materials and the Bio-Rad Liquicheck Diabetes control materials were previously reviewed in k070546 and k052838 respectively.
### Open Vial Stability:
The open-vial stability study protocols and acceptance criteria were reviewed and found acceptable. Results support the open vial stability claim for reconstituted D-100 HbA1c Calibrator Pack of 24 hours when stored at 2-8 °C.
### Closed Vial Stability:
The closed-vial stability study protocols and acceptance criteria were reviewed and found acceptable. Results support a closed vial stability claim for reconstituted D-100 Calibrator Pack of 24 months when stored at 2-8 °C. Real time stability studies are ongoing.
d. Detection limit:
Not applicable.
e. Analytical specificity:
i. Endogenous Interference
An Endogenous Interference studies was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. This study assessed common or known endogenous substances that could interfere with the D-100 HbA1c test. Potentially interfering endogenous substances were analyzed by spiking whole blood samples with HbA1c values of ~6.5% HbA1c and ~8% HbA1c with potentially interfering substances to create test samples. Ten replicates of each test and reference sample
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(sample containing no potential interferent) were analyzed and the difference in HbA1c value between the mean of test and reference replicates was determined. Significant interference was defined by the sponsor as $\geq 7\%$ change in HbA1c value of the mean of the test samples relative to mean of the reference samples.
The following endogenous substances showed no significant interference at the concentrations described below:
Table 10: Endogenous interfering substances tested
| Endogenous Substance | Concentration (Conventional Units) |
| --- | --- |
| Lipemia (Intralipid) | 6000 mg/dL |
| Conjugated bilirubin | 60 mg/dL |
| Unconjugated bilirubin | 60 mg/dL |
| Glucose | 2000 mg/dL |
| Rheumatoid factor | 750 IU/mL |
| Total protein | 21 g/dL |
ii. Drug Interference
A Drug Interference studies was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. This study assessed common or known drugs that could interfere with the D-100 HbA1c Test. Two whole blood sample pools at $\sim 6.5\%$ HbA1c (Low) and $\geq 8.0\%$ HbA1c (High) were spiked with each potentially interfering drug to create test samples. Ten replicates of each test and reference sample (sample containing no potential interferent) were analyzed and the difference in HbA1c value between the mean of test and reference replicates was determined. Significant interference was defined by the sponsor as $\geq 7\%$ change in HbA1c value of the mean of the test samples relative to mean of the reference samples.
The following endogenous substances showed no significant interference at the concentrations described below:
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Table 11: Highest tested non-interfering drug concentrations
| Drug Name | Concentration |
| --- | --- |
| Acetylcysteine | 166 mg/dL |
| Ampicillin-Na | 1000 mg/dL |
| Ascorbic acid | 300 mg/dL |
| Cefoxitin | 2500 mg/dL |
| Heparin | 5000 U/L |
| Levodopa | 20 mg/dL |
| Methyldopa | 20 mg/dL |
| Metronidazole | 200 mg/dL |
| Doxycyclin | 50 mg/dL |
| Acetylsalicylic acid | 1000 mg/dL |
| Rifampicin | 64 mg/L |
| Cyclosporine | 5 mg/L |
| Acetaminophen | 200 mg/L |
| Ibuprofen | 500 mg/L |
| Theophylline | 100 mg/L |
| Phenylbutazone | 400 mg/L |
iii. Cross Reactivity with Hemoglobin Derivatives
Hemoglobin Derivatives Interference studies was performed based on CLSI EP07-A2, Interference Testing in Clinical Chemistry. These studies study assessed potential interference from Acetylated hemoglobin (Hb), Carbamylated hemoglobin (Hb) and Labile HbA1c. Two whole blood sample pools at ~6.5% HbA1c (Low) and ≥8.0% HbA1c (High) were spiked with each potentially interfering hemoglobin derivative. Serial dilutions of each high concentration test sample were prepared, resulting in five or six levels of each potentially interfering hemoglobin derivative. Ten replicates of each level and reference sample (sample containing no potential interferent) were analyzed and the difference in HbA1c value between the mean of each level and the reference was determined. Significant interference was defined by the sponsor as ≥7% change in HbA1c value of the mean of the test samples relative to mean of the reference samples. The test result conclusions are as follows:
- Carbamylated Hb - (up to 5% Carbamylated Hb) does not interfere with this assay.
- Labile A1c- (up to 7% Labile A1c, ≈1200mg/dL glucose) does not interfere with
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this assay.
- Acetylated Hb- (up to $50\mathrm{mg / dL}$ acetylsalicylic acid) does not interfere with this assay.
Results showed there was no cross reactivity with these substances at physiological levels and the labeling states that at physiologically occurring concentrations there is no interference from labile A1c, carbamylated hemoglobin or acetylated hemoglobin.
# iv. Hemoglobin Variant Interference
A Hemoglobin Variant Interference study was performed using a total of 135 samples known to contain hemoglobin variants S, C, E, D, A2 and F. 78 whole blood patient samples containing an HbA1c $\sim 6.5\%$ and $\sim 9.0\%$ and the appropriate hemoglobin variant concentrations at levels typically found in heterozygous individuals (up to $30\%$ to $40\%$ ) were tested using the D-100 HbA1c Test on the D-1000 Hemoglobin Testing System and compared to results obtained by a reference method that has been demonstrated to be free from interference with the hemoglobin variant being tested (Trinity Biotech Ultra2 A1c and Premier Hb9210 analyzers). The following tables show the samples that were measured (Table 12) and the range of biases that were observed (Table 13) for Hemoglobin Variants S, C, D and E, and the mean biases with 2 standard deviations observed for Hemoglobin Variants F and A2 :
Table 12: Hemoglobin variant samples ranges
| Hemoglobin Variant | Number of samples | Range in % Abnormal Variant | Range in % HbA1c Concentration |
| --- | --- | --- | --- |
| HbS | 20 | 28.7 - 40.2 | 5.6 - 9.6 |
| HbC | 20 | 34.4 - 44.1 | 5.0 - 10.7 |
| HbD | 20 | 36.6 - 43.4 | 5.8 - 8.6 |
| HbE | 20 | 25.5 – 32.5 | 5.9 - 8.3 |
| HbF | 30 | 4.1-30.2 | 4.4-14.4 |
| HbA2 | 25 | 5.0-13.3 | 5.0-14.5 |
Table 13: Relative Bias Summary
| Hemoglobin Variant | Relative % Bias (Range of % Bias) observed relative to Reference Method | |
| --- | --- | --- |
| | HbA1c ~6.5% A1c | HbA1c ~9% A1c |
| HbS | -0.6 (-5.8 to 5.5) | -1.5 (-3.3 to -0.1) |
| HbC | -1.3 (-4.0 to 1.3) | -3.9 (-5.5 to -2.4) |
| HbD | -4.7 (-6.7 to -1.1) | -4.4 (-6.3 to -.2.4) |
| HbE | -2.7 (-6.7 to 1.6) | -1.3 (-2.0 to -0.6) |
| HbA2 | -1.3 (-5.1 to 0.5) | 3.4 (2.8 to 4.1) |
| HbF | -2.3 (-4.1 to -0.7) | -3.5 (-4.2 to -2.8) |
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 ( $\leq 40.2\%$ ), HbC ( $\leq 42.1\%$ ), HbD ( $\leq 42.6\%$ ), HbE ( $\leq 32.5\%$ ), HbF
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$(\leq 30.2\%)$ and HbA2 $(\leq 13.3\%)$
f. Assay cut-off:
Not applicable.
# 2. Comparison studies:
a. Method comparison with predicate device:
A Method comparison study was performed per CLSI EP09-A2 IR, Method Comparison and Bias Estimation Using Patient Samples. 129 variant-free whole blood $\mathrm{K}_{3}$ -EDTA samples, including 4 spiked samples (2 samples at HbA1c $<5\%$ and 2 samples at HbA1c $>9\%$ ), ranging from $3.4\%$ to $19.2\%$ HbA1c were evaluated using D-100 HbA1c Test on the D-100 HbA1c Testing System. Samples were tested in singlicate over four days. The results were compared to testing performed at a NGSP Secondary Reference Laboratory using a previously cleared HPLC HbA1c assay method (Tosoh G8 HPLC analyzer). To support the diagnostic claim, the distribution of samples spanned around the clinical decision point as follows in the table below.
Table 14: Sample distribution in the method comparison study
| Hemoglobin A1c level | Number of samples | % of Samples |
| --- | --- | --- |
| ≤ 5% | 6 | 4.7 |
| 5 – 6% | 17 | 13.2 |
| 6 – 6.5% | 31 | 24.0 |
| 6.5 – 7% | 33 | 25.6 |
| 7 – 8% | 20 | 15.5 |
| 8 – 9% | 11 | 8.5 |
| > 9% | 11 | 8.5 |
| Total samples | 129 | 100 |
Bias between Candidate and NGSP method
Deming (weighted) and Passing-Bablok regression analyses were performed for the D-100 Hemoglobin Testing System versus the NGSP SRL reference method.
Table 15: Summary of Method Comparison Results
| | y-Intercept | 95% CI | Slope | 95% CI |
| --- | --- | --- | --- | --- |
| Deming | 0.0223 | -0.0684 - 0.1131 | 0.9867 | 0.9736 – 0.9999 |
| Passing-Bablok | -0.0091 | -0.0803 – 0.0763 | 0.9909 | 0.9789 – 1.0026 |
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Scatter Plot using Deming Fit, %HbA1c, NGSP SRL vs. D-100 HbA1c
Scatter Plot using Passing-Bablok Fit, %HbA1c, NGSP SRL vs. D-100 HbA1c
The following biases between the D-100 HbA1c Test run on the D-100 HbA1c Testing System versus the NGSP Reference Method (Tosoh G8 HPLC analyzer) were observed:
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Table 16: Deming Regression Analysis (NGSP):
| % HbA1c Decision Level | Bias (in % HbA1c) | % Bias |
| --- | --- | --- |
| 5.0 | -0.044 | -0.88 |
| 6.5 | -0.064 | -0.98 |
| 8.0 | -0.084 | -1.05 |
| 12.0 | -0.136 | -1.13 |
Table 17: Passing-Bablok Regression Analysis (NGSP):
| % HbA1c Decision Level | Bias (in % HbA1c) | % Bias |
| --- | --- | --- |
| 5.0 | -0.055 | -1.09 |
| 6.5 | -0.068 | -1.05 |
| 8.0 | -0.082 | -1.02 |
| 12.0 | -0.118 | -0.99 |
Total Error Near the Cutoff
Using the results of bias estimation (%Bias) in the method comparison study and precision estimates in the precision study, Total Error (TE) at four HbA1c concentrations (5.0%, 6.5%, 8.0% and 12.0%) was calculated as follows: %TE =|%Bias| + 1.96 *%CV*(1+%Bias).
The results are presented in the tables below.
Table 18: Total Error
| Decision Level (% HbA1c) | % Bias | % CV | % TE |
| --- | --- | --- | --- |
| 5.0 | -0.84 | 1.7 | 4.2 |
| 6.5 | -0.98 | 1.5 | 3.9 |
| 8.0 | -1.11 | 1.3 | 3.6 |
| 12.0% | -1.57 | 1.2 | 3.9 |
b. Matrix comparison:
A matrix study was performed to determine the suitability of K2-EDTA, K3-EDTA, potassium oxalate/sodium fluoride, sodium citrate, sodium heparin and lithium heparin anticoagulants used with fresh whole blood for use with the D-100 HbA1c Test with the D-100 HbA1c Hemoglobin Testing System. Specimens with concentration values spanning 3.5 to 20.0% HbA1c were collected from a total of 43 different donors. The 43 paired samples included the following sample tube types: (1) K2 EDTA, (2) K3 EDTA, (3) Potassium Oxalate/Sodium Fluoride, (4) Sodium Citrate, (5) Sodium Heparin, and (6) Lithium Heparin.
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Three samples were altered to cover the extreme ends of the measuring range. All samples were compared to results from whole blood in $\mathrm{K}_3$ EDTA; regression results are as follows.
Table 19: Matrix comparison regression results
| Sample Type | N | K3-EDTA Sample Ranges | Slope | 95% CI of Slope | | Intercept | 95% CI of Intercept | | R2 |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | Lower | Upper | | Lower | Upper | |
| K2-EDTA | 43 | 3.53-20.06 | 0.9929 | 0.9861 | 0.9997 | 0.0270 | -0.0204 | 0.0745 | 0.9995 |
| K oxalate / NaF | 43 | 3.53-20.06 | 0.9927 | 0.9853 | 1.0002 | 0.0761 | 0.0243 | 0.1279 | 0.9994 |
| Na citrate | 43 | 3.53-20.06 | 1.0084 | 1.0017 | 1.0152 | -0.0429 | -0.0899 | 0.0040 | 0.9996 |
| Na heparin | 43 | 3.53-20.06 | 0.9972 | 0.9907 | 1.0037 | 0.0131 | -0.0322 | 0.0584 | 0.9996 |
| Li heparin | 43 | 3.53-20.06 | 0.9959 | 0.9905 | 1.0013 | 0.0103 | -0.0273 | 0.0479 | 0.9997 |
In this Matrix Comparison study, the following tube types were under evaluation and were found to be acceptable for use with the D-100 HbA1c Test when using the D-100 HbA1c Hemoglobin Testing System :
- $\mathrm{K}_{2}$ -EDTA tubes with fresh whole blood
- $\mathrm{K}_{3}$ -EDTA tubes with fresh whole blood
- potassium oxalate/sodium fluoride with fresh whole blood
- sodium citrate with fresh whole blood
- sodium heparin with fresh whole blood
- lithium heparin with fresh whole blood
# 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:
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Not applicable.
# 5. Expected values/Reference range:
The sponsor provided the following expected values in their labeling:
The following HbA1c ranges recommended by the American Diabetes Association (ADA) may be used as an aid in the diagnosis of diabetes mellitus:
| Hemoglobin A1c | | Suggested Diagnosis |
| --- | --- | --- |
| NGSP % | IFCC mmol/mol | |
| ≥6.5 | ≥48 | Diabetic4-6 |
| 5.7-6.4 | 39-47 | Pre-Diabetic4 |
| <5.7 | <39 | Non-Diabetic |
The expected HbA1c range for non-diabetic adults is $4 - 6\%$ .
4. American Diabetes Association. Diagnosis and Classification of Diabetes Mellitus. Diabetes Care 2010, 33 (Suppl. 1), S62-S69.
5. International Expert Committee. Report on the Role of the A1c Assay in the Diagnosis of Diabetes. Diabetes Care 2009, 32 (7), 1327-1334.
6. World Health Organization. Use of Glycated Haemoglobin (HbA1c) in the Diagnosis of Diabetes Mellitus. http://www.who.int/diabetes/publications/diagnosis_diabetes2011/en/ (accessed May 2015).
Clinical Diabetes. Your A1C Results: What Do They Mean? http://clinical.diabetesjournals.org/content/24/1/9.full.pdf+ (accessed November 2015).
# N. Instrument Name:
D-100 Hemoglobin Testing System
# O. System Descriptions:
# 1. Modes of Operation:
Does the applicant's device contain the ability to transmit data to a computer, webserver, or mobile device?
Yes ☐ X ☐ or No ☐
Does the applicant's device transmit data to a computer, webserver, or mobile device
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using wireless transmission?
Yes _______ or No ☐ X
2. Software:
FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:
Yes ☐ X or No _______
3. Specimen Identification:
Sample identification is via barcoding or manual entry.
4. Specimen Sampling and Handling:
A study was conducted to show the stability of samples collected in: (1) K₂-EDTA, (2) K₃-EDTA, (3) Potassium Oxalate/Sodium Fluoride, (4) Sodium Citrate, (5) Sodium Heparin, and (6) Lithium Heparin. Study protocols and acceptance criteria were reviewed and found to be acceptable to support storage of samples in all of these matrices at 15-35°C for 1 day, at 2-8°C for 7 days, at -20°C for 7 days and at -70°C for 6 months. Additional real-time studies of sample stability at -70°C are ongoing.
A study was conducted to show the stability of pre-diluted samples at system operating temperatures. Study protocols and acceptance criteria were reviewed and found to be acceptable to support storage of pre-diluted samples for 3 hours at 15-35°C.
5. Calibration:
Calibration must be performed once following the installation of every new analytical cartridge. Additional calibration may be performed at the discretion of the laboratory. Calibration is performed using the D-100 HbA1c Calibrator Pack.
6. Quality Control:
Quality control of the D-100 Hemoglobin Testing System can be conducted using commercially available Bio-Rad Lyphocheck Diabetes control materials and Bio-Rad Liquicheck Diabetes control materials; these were previously cleared in k070546 and k052838 respectively. Labeling for the device states that in keeping with good laboratory practice, at least one diabetic and one non-diabetic control specimen should be tested each day patient samples are tested and that each laboratory should establish its own guidelines for corrective action to be taken if the expected control values are not obtained.
P. Other Supportive Instrument Performance Characteristics Data Not Covered In The
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"Performance Characteristics" Section above:
Q. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
R. Conclusion:
1. The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
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