12000949 D-10™

Hemoglobin A1c
Program

Instructions For Use

400

0459

US:

0001478
This IFU is compliant with Regulation (EU) 2017/746 (IVDR).
Additional languages are available from [Link].
Significant changes are highlighted!
+ 800 135 79 135
1 855 805 85 39 (CA)
1 855 236 09 10 (US)

January 2024 UNITED STATES, Bio-Rad Laboratories, Inc., 4000 Alfred Nobel Drive,
LB002933revA Hercules, CA 94547
FRANCE, Bio-Rad, 3 boulevard Raymond Poincaré, 92430
Marnes‑la‑Coquette
D-10™ Hemoglobin A1c Program

Symbols Lexicon

Manufacturer Date of Manufacture

CE Marking - Regulation
(EU) 2017/746 (IVDR)

Authorized Representative Lot Number Use by

in the European Union

For In Vitro Diagnostic Use Catalog Number

Temperature Limit

Consult Instructions for Use For use with

Number of Tests

Prescription Use Only <0.05% Analytical Cartridge

Elution Buffer 1 Elution Buffer 2 Calibrator Level 1

Calibrator Level 2 Calibrator Diluent Calibrator/Diluent Set

Calibrator Set CD-ROM Deionized Water

Reconstitute with Resin Sample Vials

Sodium Azide Wash/Diluent Solution Whole Blood Primer

PRODUCT SAFETY INFORMATION

Whole Blood Primer and Hemoglobin A1c Calibrator 1 and 2
WARNING: This product can expose you to Tobramycin, which is known to the State of California to cause birth
defects or other reproductive harm.

Hemoglobin A1c Calibrator 1 and 2

EUH208—Contains Gentamicin, sulfate (salt). May produce an allergic reaction.

ii Instructions For Use LB002933revA

 D-10™ Hemoglobin A1c Program

Table of Contents
Product Safety Information.................................................................................................................. ii
Intended Use........................................................................................................................................3
Summary and Explanation of the Test.................................................................................................3
Principle of the Procedure...................................................................................................................4
Test Components.................................................................................................................................4
Additional Items Available from Bio-Rad.............................................................................................5
Additional Required Items Not Available from Bio-Rad.......................................................................5
Precautions/Warnings..........................................................................................................................6
Specimen Collection and Handling......................................................................................................7
Specimen Type............................................................................................................................7
Specimen Additives, Preservatives.............................................................................................7
Specimen Storage.......................................................................................................................7
Specimen Preparation.................................................................................................................7
Specimen Shipping.....................................................................................................................8
Preparation and Storage of Reagents.................................................................................................8
Indications of Instability or Deterioration of Reagents.........................................................................9
Procedure.............................................................................................................................................9
Method Selection........................................................................................................................9
Installing a New Kit Lot (Update Kit CD-ROM)...........................................................................9
Analytical Cartridge Priming Procedure...................................................................................10
Calibration.................................................................................................................................10
QC Requirements.....................................................................................................................10
Routine Run..............................................................................................................................10
Installing a Supplemental Reagent Pack..................................................................................11
Certification/Traceability to Reference Material and Method....................................................11
Guidelines for the Interpretation of Results.......................................................................................11
Limitations of the Procedure..............................................................................................................12
Sample Dilution.........................................................................................................................12
Special Considerations.............................................................................................................12
Hemoglobin Variants.................................................................................................................13
Interfering Substances..............................................................................................................13
Expected Values/Reference Range...................................................................................................14
Diagnosis of Diabetes...............................................................................................................14
Monitoring HbA1c in Diabetic Patients.......................................................................................14
Performance Characteristics..............................................................................................................15
Precision....................................................................................................................................15
Accuracy ...................................................................................................................................18
Linearity.....................................................................................................................................18

LB002933revA Instructions For Use 1

D-10™ Hemoglobin A1c Program

Sample Reports.................................................................................................................................19
Trademark Information.......................................................................................................................28
References.........................................................................................................................................28

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 D-10™ Hemoglobin A1c Program

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

SUMMARY AND EXPLANATION OF THE TEST

Diabetes mellitus is a condition characterized by hyperglycemia resulting from the body’s inability to use blood
glucose for energy. In Type 1 diabetes, the pancreas no longer makes insulin and therefore, blood glucose
cannot enter the cells to be used for energy. In Type 2 diabetes, either the pancreas does not make enough
insulin or the body is unable to use insulin correctly.1 The direct and indirect effects of hyperglycemia on the
human vascular system are the major source of morbidity and mortality in both Type 1 and Type 2 diabetes.
These effects include macrovascular complications (coronary artery disease, peripheral arterial disease, and
stroke) and microvascular complications (diabetic nephropathy, neuropathy, and retinopathy).2 Diabetes mellitus
affects >8% of the world population.3
HbA1c testing has been recommended for the diagnosis of Type 2 diabetes by the International Expert
Committee (IEC), the American Diabetes Association (ADA), and the World Health Organization (WHO),
which recommend a diagnostic threshold of ≥6.5% (≥48 mmol/mol) HbA1c.4–6 HbA1c testing has also been
recommended for the identification of individuals at increased risk for developing diabetes (pre-diabetic). The
ADA has defined the HbA1c range for pre-diabetes as 5.7–6.4% (39–47 mmol/mol).4 Detection and treatment of
pre-diabetes may reduce or eliminate the risk of developing Type 2 diabetes and related complications.
Therapy for diabetes requires the long-term maintenance of a blood glucose level as close as possible to
a normal level, minimizing the risk of long-term vascular consequences.7,8 A single fasting blood glucose
measurement is an indication of the patient’s immediate past condition (hours), but may not represent the true
status of blood glucose regulation.9,10 The measurement of hemoglobin A1c (HbA1c) every two to three months
has been accepted as a measure of glycemic control in the care and treatment of patients with diabetes mellitus.
HbA1c, the glycohemoglobin of interest, is formed in two steps by the nonenzymatic glycation of HbA. The
first step is the formation of an unstable aldimine (labile A1c, or pre-A1c), a reversible reaction between the
carbonyl group of glucose and the N terminal valine of the β-chain of hemoglobin. Labile A1c formation is directly
proportional to the blood glucose concentration. During red blood cell circulation, some of the labile A1c is
converted (Amadori rearrangement) to form a stable ketoamine, HbA1c.11
The D-10 Hemoglobin A1c Program is based on chromatographic separation of HbA1c on a cation exchange
cartridge. Separation is optimized to minimize interferences from hemoglobin variants, labile A1c, and
carbamylated hemoglobin. Please refer to Limitations of the Procedure for more information. The D-10
Hemoglobin A1c Program also offers automatic sampling from a primary whole blood tube, followed by sample
dilution, and an analysis time of three minutes per sample.

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D-10™ Hemoglobin A1c Program

PRINCIPLE OF THE PROCEDURE

The D-10 Hemoglobin A1c Program utilizes principles of ion-exchange high-performance liquid chromatography
(HPLC). The samples are automatically diluted on the D-10 and injected into the analytical cartridge. The D-10
delivers a programmed buffer gradient of increasing ionic strength to the cartridge, where the hemoglobins are
separated based on their ionic interactions with the cartridge material. The separated hemoglobins then pass
through the flow cell, where changes in the absorbance at 415 nm are measured.
The D-10 software collects raw data from each analysis and calculates HbA1c values based on a bi-level
calibration curve. A sample report and a chromatogram are generated for each sample. The A1c peak is shaded.
This area is calculated using an exponentially modified Gaussian (EMG) algorithm.
The D-10 Hemoglobin A1c Program is for use only with the D-10 Hemoglobin Testing System.

TEST COMPONENTS
12000949, D-10 Hemoglobin A1c Program
The kit contains supplies for 400 tests:

Quantity Description
220-0110* 2 each Elution Buffer 1. Each bottle contains 2000 mL of a Bis-Tris/Phosphate
buffer. Contains <0.05% sodium azide as a preservative.
220-0111* 1 each Elution Buffer 2. Each bottle contains 1000 mL of a Bis-Tris/Phosphate
buffer. Contains <0.05% sodium azide as a preservative.
220-0112* 1 each Wash/Diluent Solution. Each bottle contains 1600 mL of deionized water
with <0.05% sodium azide as a preservative.
12005707* 1 each Analytical Cartridge. Cation exchange cartridge (400 tests),
4.0 mm ID x 40 mm.
12002715* 1 each CD-ROM with D-10 Hemoglobin A1c "Update Kit" program parameters.
12005706* 1 each Calibrator/Diluent Set. One set consisting of 3 vials of Calibrator Level 1,
3 vials of Calibrator Level 2, and 1 bottle of Calibrator Diluent. The
calibrator vials contain lyophilized human red blood cell hemolysate with
gentamicin, tobramycin, and EDTA as preservatives. Reconstituted volume
is 7 mL per vial.
Calibrator Diluent contains 100 mL of deionized water with <0.05% sodium
azide as a preservative.
220-0148* 4 each Whole Blood Primer. Each vial contains lyophilized human red blood
cell hemolysate with gentamicin, tobramycin, and EDTA as preservatives.
Reconstituted volume is 1.0 mL per vial.
12005138 1 each Sample Vials. 50 polypropylene microvials with pierceable caps, 1.5 mL.

* Components are not available for individual sale.

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 D-10™ Hemoglobin A1c Program

220-0109, D-10 Hemoglobin A1c Supplemental Reagent Pack

The reagent pack is used as a supplement to the D-10 Hemoglobin A1c Program, providing low-test-volume
users with buffers and wash/diluent solution to achieve 400 tests.

Quantity Description
220-0110* 2 each Elution Buffer 1. Each bottle contains 2000 mL of a Bis-Tris/Phosphate
buffer. Contains <0.05% sodium azide as a preservative.
220-0111* 1 each Elution Buffer 2. Each bottle contains 1000 mL of a Bis-Tris/Phosphate
buffer. Contains <0.05% sodium azide as a preservative.
220-0112* 1 each Wash/Diluent Solution. Each bottle contains 1600 mL of deionized water
with <0.05% sodium azide as a preservative.

* Components are not available for individual sale.

NOTE: The D-10 calculates buffer levels and waste level assuming test volumes of 200 or more tests per month
and 4 or more samples per run. Low-test-volume users will run out of buffers and fill the external waste tank
before the applicable warning messages are displayed. Visually check the buffer levels and waste tank level prior
to each run.

ADDITIONAL ITEMS AVAILABLE FROM BIO-RAD

Description
220-0297 Microvial Adapters, 10 x 1.5 mL
220-0468 D-10 Thermal Paper, 10 rolls, for KyoSpeed 2 Printer
740 Lyphochek™ Diabetes Control Bilevel, 6 x 0.5 mL
740X Lyphochek™ Diabetes Control Bilevel MiniPak, 2 x 0.5 mL
12000070 Lyphochek™ Hemoglobin A1c Linearity Set (1 each of 6 levels), 6 x 0.5 mL
12011343 Liquichek™ Diabetes Control, Level 1, 6 x 1.0 mL
12011344 Liquichek™ Diabetes Control, Level 2, 6 x 1.0 mL
12011345 Liquichek™ Diabetes Control, Level 3, 6 x 1.0 mL
12011334 Liquichek™ Diabetes Control, Trilevel MiniPak, 3 x 1.0 mL

ADDITIONAL REQUIRED ITEMS NOT AVAILABLE FROM BIO-RAD

Pipettes, 5 µL, 0.5 mL, 1 mL, 1.5 mL, 7 mL
Deionized Water

LB002933revA Instructions For Use 5

D-10™ Hemoglobin A1c Program

PRECAUTIONS/WARNINGS
• For in vitro diagnostic use by a professional user in a laboratory environment only.
• This test kit should be handled only by qualified personnel trained in laboratory procedures and familiar with
the potential hazards.
• This product contains human source materials. Consider any materials of human origin as infectious and
handle them using typical biosafety procedures.
• Wear personal protective equipment while handling all reagents and samples and while operating the system.
• Dispose of all waste in accordance with applicable national and/or local regulations.
• Some reagents contain sodium azide, which may react with copper or lead plumbing to form explosive metal
azides. Use caution in disposing of these reagents. If disposing to drain, flush with large volumes of water to
prevent azide buildup.
• Waste material containing patient samples or biological products should be considered biohazardous when
disposing or treating.
• Chemical reagents should be handled in accordance with Good Laboratory Practices.
• Biological spills: Human source material spills should be treated as potentially infectious. Spills not containing
acid should be immediately decontaminated, including the spill area, materials and any contaminated
surfaces or equipment, with an appropriate chemical disinfectant that is effective for the potential biohazards
(commonly a 1:10 dilution of household bleach, 70–80% Ethanol or Isopropanol, an iodophor (such as 0.5%
Wescodyne Plus, etc.), and wiped dry.
• Do not interchange vial or bottle caps and stoppers; this will lead to cross-contamination of reagents. Never
mix the contents from different bottles of the same reagent. Doing so may lead to reagent contamination and
compromise the performance of the product.
• Each unit of whole blood used in the manufacture of the calibrators and whole blood primer was tested by
FDA‑accepted methods and found non-reactive for HIV-1, HIV-2, Hepatitis B (HBV), Hepatitis C (HCV), and
syphilis. No test method can offer complete assurance that products containing human source materials
will be absent of these and other infectious agents. In accordance with good laboratory practice, all human
source material should be considered potentially infectious for all infectious agents; therefore, handle the
calibrators and whole blood primer with the same precautions used with patient specimens.
• Adherence to the protocol specified herein is necessary to ensure proper performance of this product.
• Do not use the Calibrator Diluent for prediluting patient samples.
• For complete details on safe reagent handling, refer to the Safety Data Sheets (SDS) available at
[Link].
• Bio-Rad has established assay performance according to the specimen handling and storage parameters
described in this IFU. If a laboratory uses handling and storage criteria outside of the guidance listed here,
it is the responsibility of the individual laboratory to use all available references and/or its own studies to
determine specific stability criteria for their procedures.
• The user should not make any decision of medical relevance without first consulting the appropriate
healthcare professional in the member state(s) where the device is placed on the market.
• For a patient/user/third party in the European Union and in countries with identical regulatory regime
requirements (Regulation 2017/746/EU on In vitro Diagnostic Medical Devices): If during the use of this
device or as a result of its use, a serious incident occurs, please report it to Bio-Rad Laboratories and/or its
authorized representative and to your national Competent Authority.

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 D-10™ Hemoglobin A1c Program

SPECIMEN COLLECTION AND HANDLING

Specimen Type
Whole blood.

Specimen Additives, Preservatives

The whole blood specimens should be collected in vacuum collection tubes containing K2-EDTA or K3-EDTA.
A matrix comparison study was performed to evaluate blood collection tube anticoagulants/preservatives for use
with the test. Whole blood samples from 44 patients with HbA1c concentrations spanning the reportable range of
the test were collected in the evaluation tube type and the comparative tube type (K3-EDTA). The results of the
matrix comparison study are summarized in Table 1.

Anticoagulant/
Slope Intercept R2
Preservative
K2-EDTA 1.0077 −0.0383 0.9990
Table 1: Regression Analysis of % HbA1c for Evaluation Tubes vs K3-EDTA Tubes

Specimen Storage
Whole blood specimens may be stored as follows:
• 3 days at room temperature (15–30 °C)
• Up to 7 days at 2–8 °C
• Up to 12 months at −70 °C

Specimen Preparation
Whole Blood Samples:
• Allow sample tubes to reach room temperature (15–30 °C) before performing the test.
• No sample preparation is required. Mixing has no impact on HbA1c values as long as the total area is in
range.
NOTE: Studies indicate that the use of MONOJECT ( 8881311446, Covidien, Mansfield, MA, 02048 USA)
collection tubes on the D-10 Hemoglobin Testing System can result in elevated total area. In some cases, blood
collects under the cap and causes oversampling, resulting in high total area that exceeds the acceptable range.
To use this tube type, users must either manually dilute the sample or ensure that blood does not collect under
the cap before loading the sample on the D-10 Hemoglobin Testing System. Results with total area within the
acceptable range are reportable.

Prediluted Samples:
If the sample is in an abnormal size/type tube, or if there is less than 2.0 mL of sample in the tube, then the
sample must be prediluted 1:300 prior to analysis:
1. Before pipetting, thoroughly mix the sample by gently inverting the tube.
2. To predilute, pipet 1.5 mL of Wash/Diluent Solution into a labeled 1.5 mL microvial, followed by 5 μL of the
whole blood sample.
3. Cap the microvial and mix thoroughly.
4. Use a microvial adapter for 1.5 mL microvials.
NOTE: Samples with total areas outside of the expected range should be rediluted and rerun to achieve values
within the 1.0–5.0 million total area count range.

LB002933revA Instructions For Use 7

D-10™ Hemoglobin A1c Program

Specimen Shipping
All samples of human origin must be shipped in accordance with national and international transportation
regulations.
The collected whole blood samples must be transported at 1–10 °C. If a delay in testing or shipping is expected,
store the specimens at 2–8 °C. The blood collection, transport, and testing must total ≤7 days. If the samples will
not be tested within 7 days of collection, samples may be stored frozen at −70 °C upon receipt. Samples can
withstand up to 3 freeze-thaw cycles.

PREPARATION AND STORAGE OF REAGENTS

To install or change Elution Buffers, Wash/Diluent Solution, and Analytical Cartridge, follow the procedures
described in the D-10 Operation Manual.

Elution Buffers and Wash/Diluent Solution

• Allow the Elution Buffers and Wash/Diluent Solution to reach room temperature (15–30 °C) before performing
the test. Mix each bottle by gently inverting prior to installation.
• The Elution Buffers and Wash/Diluent Solution are stable until the expiration date when stored unopened at
15–30 °C. After opening the bottles, these reagents are stable for 8 weeks when stored at 15–30 °C.
• With a new kit, install one bottle of each reagent and follow the procedure for Installing a New Kit Lot in the
Procedure section. After 200 injections, install a fresh bottle of Elution Buffer 1. Reset the volume in the LOT
INFO/Buffer 1 screen after installing this reagent.
NOTE: When using the optional D-10 Rack Loader, the two bottles of Elution Buffer 1 are installed
simultaneously. Manually resetting the volume is not required.
• The Wash/Diluent Solution is interchangeable between kit lots.
• Do not mix or use reagents from different lots within a test run.

Whole Blood Primer

• Use fresh aliquots of Whole Blood Primer when installing a new cartridge.
• The Whole Blood Primer is stable until the expiration date when stored unopened at 2–8 °C.
• The Whole Blood Primer is provided in lyophilized form for increased stability.
1. Reconstitute each Whole Blood Primer by adding 1.0 mL of deionized water to each vial.
2. Replace the vial stoppers and allow vials to stand for 10 minutes at 15–30 °C.
3. Swirl gently to dissolve and ensure complete mixing.
• The reconstituted Whole Blood Primer is stable for 1 day when stored at 2–8 °C.
• The Whole Blood Primer is interchangeable between lots.

Calibrator/Diluent Set
• The Calibrator/Diluent Set is stable until the expiration date when stored unopened at 2–8 °C.
• The Calibrator Diluent is ready to use. The Diluent is stable for 8 weeks, after opening the bottle, when stored
at 2–8 °C. The Diluent is interchangeable between lots.
• The Calibrators are provided in lyophilized form for increased stability.
1. Using a volumetric pipette, reconstitute each Calibrator by adding 7 mL of cold Calibrator Diluent to each
vial.
2. Replace the vial stoppers and allow vials to stand 5–10 minutes.
3. Swirl gently to dissolve.

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 D-10™ Hemoglobin A1c Program

• The reconstituted Calibrators are stable for 7 days when stored capped at 2–8 °C.
• See the value card included with the current lot of calibrators for value assignment. Values are entered
automatically using the Update Kit CD-ROM. Values must be entered manually if a different lot is being
used; see the D-10 Operation Manual to manually enter values in the LOT INFO/Calibrator 1 and LOT INFO/
Calibrator 2 screens.

Extracted Standards
This HPLC method does not use extracted standards.

Controls
Reconstitute and store the controls according to the manufacturer’s package insert. If no dilution instructions are
provided, predilute the controls as directed in the Specimen Preparation section of these Instructions For Use.

Analytical Cartridge
The Analytical Cartridge should be stored at 15–30 °C. The Analytical Cartridge is stable for 8 weeks or
400 tests when installed on the instrument.

INDICATIONS OF INSTABILITY OR DETERIORATION OF REAGENTS

• If reagents were frozen during shipment, mix each bottle by gently inverting before installing on instrument.
• Do not use any reagents which have any indications of discoloration, cloudiness, or precipitation.
• Do not use any reagents that show any signs of leakage.
• Do not use the calibrator or whole blood primer if the pellet is brown or the vial is broken. If the lyophilized
material contains insoluble matter, discard the material and reconstitute a new vial.
• Do not use the kit if the packaging of any component is damaged.
• Do not use expired reagents.

PROCEDURE
Method Selection
1. From the appropriate LOT INFO screen, select the Method field.
2. Select the desired method (HbA1c).
3. Press Exit.
4. Press Yes to confirm the method change.
5. Press Exit.
6. The selected method is indicated on the left in the status bar.
NOTE: There is no need to perform a system flush unless you are installing a new lot of reagents.

Installing a New Kit Lot (Update Kit CD-ROM)

When changing to a different lot of reagents and/or cartridge, the parameters from the matching CD-ROM must
be installed to ensure optimum performance of the program.
1. Go to any LOT INFO screen.
2. Press the Update Kit button.
3. Place the Update Kit CD-ROM in the CD drive.
4. Follow the instructions on the screen to proceed with the Update Kit Procedure.
5. Remove the CD-ROM from the drive once the procedure is completed.
LB002933revA Instructions For Use 9
D-10™ Hemoglobin A1c Program

Analytical Cartridge Priming Procedure

1. See the D-10 Operation Manual for cartridge installation instructions.
2. Pipet 1 mL of reconstituted Whole Blood Primer into a microvial.
3. Put the microvial into a microvial adapter labeled with a Primer barcode, then place the adapter into sample
rack position 1.
4. Start a run. When finished, the cartridge is ready for calibration.

Calibration
Calibration must be performed once, following the installation and priming of every new analytical cartridge.
Additional calibration may be performed at the discretion of the laboratory.
1. Prepare samples as described in Specimen Preparation section.
2. Place calibrators and controls on the D-10. Use microvial adapters for 1.5 mL microvials with calibrators and
controls. The adapters should have barcodes identifying sample type.
Sample order with calibrators:

Tube Position Adapter Label Reagent

1 Calibrator 1 Reconstituted HbA1c Calibrator Level 1 (1 mL)
2 Calibrator 2 Reconstituted HbA1c Calibrator Level 2 (1 mL)
3 A1c Low Control Prediluted Control Level 1
4 A1c High Control Prediluted Control Level 2
5–10 ---- Patient Samples

NOTE: The analyte values printed on the calibrator reports are the calibrator assigned values. In the case of
calibration failure, the reports will indicate these assigned values; however, the statement “Calibration Failed” will
be printed at the bottom of the calibrator report. If the alert setting “Stop if calibration fails” is NOT selected (see
the Alert Settings screen in the D-10 Operation Manual), the run will continue, using the calibration factors from
the previous acceptable calibration run. For troubleshooting advice, refer to the Troubleshooting section in the
D-10 Operation Manual.

QC Requirements
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 or follow locally established guidelines. Each laboratory should
establish its own guidelines for corrective action to be taken if the expected control values are not obtained.

Routine Run
Once the cartridge has been calibrated, use the following run configuration. Refer to QC Requirements for
information on control sample frequency.
Sample order without calibrators:

Tube Position Adapter Label Reagent

1 A1c Low Control Prediluted Control Level 1 (optional)
2 A1c High Control Prediluted Control Level 2 (optional)
3–10 ---- Patient Samples

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 D-10™ Hemoglobin A1c Program

Installing a Supplemental Reagent Pack

• When installing reagents from the Supplemental Reagent Pack, Buffer 1 and Buffer 2 must always be used
as a matched set. Never use a buffer bottle from the kit with a buffer bottle from the Supplemental Reagent
Pack.
• Users will need to reset the buffer volume when new bottles are installed. (Refer to the Reset Buffer Volume
Screen in the D-10 Operation Manual.)
• The Buffer and Wash/Diluent lot and expiration information must be manually entered into the system.
• When installing the second bottle of Buffer 1, reset the Buffer 1 volume.

Certification/Traceability to Reference Material and Method

The D-10 Hemoglobin A1c Program is traceable to the reference methods of both the National Glycohemoglobin
Standardization Program (NGSP) and the International Federation of Clinical Chemistry and Laboratory
Medicine (IFCC).
The D-10 Hemoglobin A1c Program is certified by the NGSP as having documented traceability to the reference
method of the Diabetes Control and Complications Trial (DCCT), where the relationship between mean blood
glucose and risk for vascular complications was established.12
The IFCC Working Group on HbA1c Standardization developed and maintains the reference measurement
procedure used as the analytical anchor for traceability of HbA1c.13 This reference method is used to assign
IFCC values to secondary reference materials that are used by manufacturers to assign product calibrator
values.14
In May 2007, the American Diabetes Association, European Association for the Study of Diabetes, International
Diabetes Federation, and IFCC issued a consensus statement on the worldwide standardization of the HbA1c
measurement. They recommended use of the IFCC SI units (mmol/mol).15
The master equations for conversion between IFCC and NGSP13 and examples of patient results are as follows:

IFCC NGSP
NGSP = (0.09148 × IFCC) + 2.152 39 mmol/mol 5.7%
IFCC = (10.93 × NGSP) − 23.50 48 mmol/mol 6.5%
64 mmol/mol 8.0%
108 mmol/mol 12.0%

GUIDELINES FOR THE INTERPRETATION OF RESULTS

Observe the following guidelines to ensure acceptable results:
1. The system must pass calibration.
2. Total area of each analysis must range from 1.0 million to 5.0 million units. Results should not be reported if
the area is outside this range.
3. The peaks A1c and A0 must be correctly identified.
4. Quality Control values should be in range.

LB002933revA Instructions For Use 11

D-10™ Hemoglobin A1c Program

5. The reportable range for HbA1c was established based on data presented in Performance Characteristics,
Linearity. If the HbA1c result falls outside the reportable range, it should not be reported.

Reportable Range
NGSP % HbA1c 3.9–18.8
IFCC mmol/mol HbA1c 19–182

6. Any sample with >15% or >140 mmol/mol HbA1c should be suspected of having a hemoglobin variant.16
7. Any sample with a combined area of ≥50% in the E-, D,S-, and/or C-window should be suspected of having
a homozygous or double-heterozygous variant, or a variant–β-thalassemia phenotype. 17,18 The HbA1c result
should not be reported for these samples.
8. For diagnosis purposes, results should be interpreted in conjunction with the patient’s medical history and
clinical findings.

Interpretation of “Unknown” and P3 Peaks

Several minor components of hemoglobin A may be resolved and listed as “Unknown” peaks in the sample
report. The number of minor “Unknown” peaks and their integrated area will vary from sample to sample. See
Figure 3 for a typical example of the integration and reporting of minor “Unknown” peaks. The largest minor
components of hemoglobin A are given designated peak window P3.19 Whole blood specimens that have been
stored or shipped outside of manufacturer guidelines may exhibit an increase in the P3 peak area. In all cases,
all components of hemoglobin A (e.g., P3, Unknown) are appropriately included in the total area to accurately
determine the relative percent of HbA1c. However, any sample with an Unknown and/or P3 peak >10% should
be suspected of having a hemoglobin variant.20

LIMITATIONS OF THE PROCEDURE

Sample Dilution
Normal total hemoglobin concentration corresponds to a total area of approximately 2.5 million units. The
required total area range for the D-10 Hemoglobin A1c Program is 1.0 million to 5.0 million units.
If the sample area is outside of the expected range, manually predilute the sample following the Specimen
Preparation guidelines. If the sample area is still outside of the expected range, the sample should be rediluted
and rerun to achieve values within the 1.0 million to 5.0 million total area count range.
NOTE: For some high total area, high HbA1c samples (e.g., 15% or 140 mmol/mol HbA1c with 5 M total area),
the A1c peak may elute outside of the established retention time window. Predilute the sample to approximately
2.5 M total area and rerun.

Special Considerations
• The HbA1c test is not intended for analysis of samples collected from newborns.
• The HbA1c test should not be used to replace glucose testing in pediatric patients, pregnant women, or
patients with Type 1 diabetes.
• 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.
• The HbA1c test 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.

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 D-10™ Hemoglobin A1c Program

• The 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.16,21−23

Hemoglobin Variants
The most common heterozygous hemoglobin variants (i.e., HbAS, HbAC, HbAD, and HbAE) do not interfere
with the test. Typical chromatograms for these variants are provided in Figures 6–9.
In the homozygous and double-heterozygous forms of variant hemoglobins (e.g., SS, CC, SC), there is no HbA
present; therefore, no HbA1c value can be determined.
The effect of common hemoglobin variants on the HbA1c result was evaluated based on the CLSI EP07-A2
guideline, “Interference Testing in Clinical Chemistry”. The relative % bias to the comparative method is
summarized in Table 2.

Relative % Bias to Comparative Method

Hemoglobin
Variant Relative % Bias (Range of Relative % Bias (Range of
% Bias) for HbA1c ~6.5% % Bias) for HbA1c ~9.0%
HbS 1.1 (−4.7 to 4.9) 0.2 (−6.1 to 5.3)
HbC 2.6 (1.4 to 5.9) −0.9 (−2.8 to 0.9)
HbD −0.6 (−3.1 to 4.7) 1.7 (−3.6 to 5.1)
HbE 0.5 (−3.3 to 6.2) 2.8 (1.2 to 5.2)
Table 2: Results of Hemoglobin Variants Interference Study
Other abnormal hemoglobin variants have not been evaluated on the D-10 Hemoglobin A1c Program. For the
confirmation of any particular hemoglobin variant, alternative methods are required.

Interfering Substances
Interference studies were conducted in accordance with the CLSI EP07-A2 guideline, “Interference Testing in
Clinical Chemistry”. Each interfering substance was evaluated using specimens with hemoglobin concentrations
of approximately 6.5% (48 mmol/mol) and ≥8.0% (≥64 mmol/mol). The following are the results of the
interference studies.
• This device has significant positive interference with fetal hemoglobin (HbF). HbA1c results are invalid
for patients with abnormal amounts of HbF, including those with known Hereditary Persistence of Fetal
Hemoglobin.

Hemoglobin F concentrations up to 10% do not interfere with the test. Any sample with HbF >10% may
result in higher than expected HbA1c values. Any sample with HbF >5% should be suspected of having a
hemoglobinopathy.24
• β-thalassemia trait, as indicated by increased HbA2 concentrations, does not interfere with the test.

Relative % Bias to Comparative Method

Hemoglobin Relative % Bias (Range of Relative % Bias (Range of
% Bias) for HbA1c ~6.5% % Bias) for HbA1c ~9.0%
HbF 3.1 (−1.5 to 8.8) −0.6 (−2.5 to 4.8)
HbA2 0.6 (−1.8 to 1.8) −1.7 (−2.8 to −0.7)

LB002933revA Instructions For Use 13

D-10™ Hemoglobin A1c Program

• Labile A1c (LA1c/CHb-1) concentrations up to 6% (or 1000 mg/dL glucose) do not interfere with the test.
• Carbamylated hemoglobin (LA1c/CHb-2) concentrations up to 3.5% (or 10 mg/dL potassium cyanate) do not
interfere with the test.
• At physiologically occurring concentrations, there is no interference from acetylated hemoglobin.25
• Common drugs at therapeutic concentrations do not interfere with the test.25 No significant interference is
observed from the following endogenous substances up to the stated concentrations:

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

EXPECTED VALUES/REFERENCE RANGE

Diagnosis of Diabetes
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%.26 Each laboratory can determine the transferability of
the expected values to its own patient population and if necessary, determine its own reference ranges.

Monitoring HbA1c in Diabetic Patients

The following HbA1c ranges may be used for interpretation of results; however, the clinician needs to work with a
patient and should consider adjusting targets if needed to improve safety and adherence. These values are for
many nonpregnant adults.23

Hemoglobin A1c
Glycemic Goal
NGSP % IFCC mmol/mol
<8 <64 Less Stringent Goal*
<7 <53 General Goal†

* M
 ay be appropriate for patients with limited life expectancy, or where the harms of treatment are greater than
the benefits.
† S
 hown to reduce microvascular complications and, if implemented soon after diagnosis of diabetes, is
associated with long-term reduction in microvascular disease.

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 D-10™ Hemoglobin A1c Program

On the basis of clinician judgment and patient preference, achievement of levels significantly lower than the goal
of 7% may be acceptable, and even beneficial, if it can be achieved safely without the risk of hypoglycemia or
other adverse effects of treatment.

PERFORMANCE CHARACTERISTICS
Precision
The precision of the D-10 Hemoglobin A1c Program was evaluated based on the CLSI EP05-A2 guideline,
“Evaluation of Precision Performance of Quantitative Measurement Methods” using a modified study design.
HbA1c results were obtained for a series of samples across the clinical range of the test by analyzing each
sample in duplicate in 2 runs per day on 3 instruments for 20 days. The study was repeated using 3 different
kit lots, yielding a total of 720 results per sample over a 60-day period. The results of the precision study are
summarized in Tables 3a (NGSP %) and 3b (IFCC mmol/mol).

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D-10™ Hemoglobin A1c Program

Instrument 1 Precision by Sample

Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(5.2%) (6.7%) (8.4%) (12.7%) (5.6%) (10.4%)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.07 1.3 0.07 1.0 0.05 0.6 0.09 0.7 0.07 1.2 0.10 0.9
Between-Run 0.00 0.0 0.00 0.0 0.01 0.2 0.06 0.5 0.00 0.0 0.00 0.0
Between-Day 0.04 0.7 0.07 1.1 0.08 1.0 0.17 1.3 0.05 0.9 0.12 1.1
Between-Lot 0.08 1.6 0.05 0.7 0.02 0.2 0.21 1.7 0.05 0.9 0.12 1.2
Total Precision 0.11 2.2 0.11 1.7 0.10 1.2 0.29 2.3 0.10 1.8 0.20 1.9
Instrument 2 Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(5.2%) (6.6%) (8.2%) (12.5%) (5.6%) (10.1%)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.04 0.9 0.03 0.5 0.04 0.5 0.06 0.5 0.04 0.8 0.04 0.4
Between-Run 0.02 0.4 0.04 0.6 0.04 0.4 0.08 0.6 0.02 0.3 0.06 0.6
Between-Day 0.04 0.8 0.05 0.8 0.07 0.8 0.13 1.1 0.06 1.0 0.08 0.8
Between-Lot 0.08 1.5 0.02 0.3 0.02 0.2 0.21 1.7 0.07 1.2 0.09 0.9
Total Precision 0.10 1.9 0.08 1.2 0.09 1.0 0.27 2.1 0.10 1.7 0.14 1.4
Instrument 3 Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(5.2%) (6.7%) (8.3%) (12.7%) (5.6%) (10.3%)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.06 1.2 0.05 0.7 0.05 0.6 0.07 0.6 0.05 0.9 0.08 0.8
Between-Run 0.00 0.0 0.02 0.3 0.03 0.4 0.07 0.6 0.00 0.0 0.05 0.5
Between-Day 0.06 1.2 0.08 1.2 0.11 1.3 0.17 1.3 0.07 1.2 0.14 1.3
Between-Lot 0.05 0.9 0.00 0.0 0.04 0.5 0.21 1.6 0.04 0.8 0.08 0.8
Total Precision 0.10 1.9 0.09 1.4 0.13 1.6 0.29 2.2 0.10 1.7 0.19 1.8
Combined Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(5.2%) (6.7%) (8.3%) (12.7%) (5.6%) (10.3%)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.06 1.2 0.05 0.8 0.05 0.6 0.08 0.6 0.06 1.0 0.08 0.7
Between-Run 0.00 0.0 0.01 0.2 0.03 0.3 0.07 0.6 0.00 0.0 0.04 0.4
Between-Day 0.05 0.9 0.07 1.0 0.09 1.1 0.16 1.2 0.06 1.0 0.12 1.1
Between-Instrument 0.00 0.0 0.05 0.8 0.08 0.9 0.00 0.0 0.00 0.0 0.14 1.3
Between-Lot 0.07 1.4 0.03 0.5 0.03 0.3 0.21 1.7 0.05 0.9 0.10 1.0
Total Precision 0.10 2.0 0.11 1.6 0.13 1.6 0.28 2.2 0.10 1.7 0.22 2.2
Table 3a: Results of Precision Study (NGSP %)

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 D-10™ Hemoglobin A1c Program

Instrument 1 Precision by Sample

Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(33 mmol/mol) (50 mmol/mol) (68 mmol/mol) (116 mmol/mol) (38 mmol/mol) (91 mmol/mol)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.76 2.3 0.76 1.5 0.53 0.8 1.01 0.9 0.77 2.0 1.08 1.2
Between-Run 0.00 0.0 0.00 0.0 0.16 0.2 0.65 0.6 0.00 0.0 0.00 0.0
Between-Day 0.40 1.2 0.81 1.6 0.92 1.3 1.82 1.6 0.56 1.5 1.31 1.4
Between-Lot 0.89 2.7 0.54 1.1 0.17 0.3 2.31 2.0 0.53 1.4 1.34 1.5
Total Precision 1.24 3.7 1.23 2.5 1.08 1.6 3.19 2.8 1.09 2.9 2.16 2.4
Instrument 2 Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(33 mmol/mol) (49 mmol/mol) (66 mmol/mol) (113 mmol/mol) (37 mmol/mol) (87 mmol/mol)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.49 1.5 0.36 0.7 0.40 0.6 0.63 0.6 0.46 1.2 0.45 0.5
Between-Run 0.20 0.6 0.42 0.9 0.38 0.6 0.88 0.8 0.19 0.5 0.66 0.8
Between-Day 0.43 1.3 0.58 1.2 0.72 1.1 1.47 1.3 0.61 1.6 0.89 1.0
Between-Lot 0.84 2.6 0.24 0.5 0.19 0.3 2.27 2.0 0.70 1.9 0.96 1.1
Total Precision 1.08 3.3 0.83 1.7 0.93 1.4 2.91 2.6 1.05 2.8 1.54 1.8
Instrument 3 Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(33 mmol/mol) (50 mmol/mol) (68 mmol/mol) (115 mmol/mol) (37 mmol/mol) (89 mmol/mol)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.70 2.1 0.51 1.0 0.56 0.8 0.80 0.7 0.57 1.5 0.87 1.0
Between-Run 0.00 0.0 0.22 0.4 0.33 0.5 0.80 0.7 0.00 0.0 0.60 0.7
Between-Day 0.66 2.0 0.85 1.7 1.20 1.8 1.81 1.6 0.73 2.0 1.51 1.7
Between-Lot 0.51 1.6 0.00 0.0 0.46 0.7 2.26 2.0 0.46 1.2 0.93 1.0
Total Precision 1.09 3.3 1.02 2.0 1.44 2.1 3.11 2.7 1.03 2.8 2.07 2.3
Combined Precision by Sample
Patient 1 Patient 2 Patient 3 Patient 4 Control 1 Control 2
Variation Source
(33 mmol/mol) (50 mmol/mol) (67 mmol/mol) (115 mmol/mol) (37 mmol/mol) (89 mmol/mol)
SD % CV SD % CV SD % CV SD % CV SD % CV SD % CV
Repeatability 0.66 2.0 0.57 1.1 0.50 0.7 0.83 0.7 0.60 1.6 0.84 0.9
Between-Run 0.00 0.0 0.13 0.3 0.31 0.5 0.78 0.7 0.00 0.0 0.48 0.5
Between-Day 0.51 1.5 0.76 1.5 0.96 1.4 1.72 1.5 0.63 1.7 1.26 1.4
Between-Instrument 0.00 0.0 0.58 1.2 0.84 1.3 0.00 0.0 0.00 0.0 1.49 1.7
Between-Lot 0.77 2.3 0.34 0.7 0.30 0.4 2.29 2.0 0.57 1.5 1.09 1.2
Total Precision 1.13 3.4 1.17 2.3 1.44 2.1 3.08 2.7 1.04 2.8 2.44 2.7
Table 3b: Results of Precision Study (IFCC mmol/mol)

LB002933revA Instructions For Use 17

D-10™ Hemoglobin A1c Program

Accuracy
The D-10 Hemoglobin A1c Program was compared to the NGSP Secondary Reference Laboratory (SRL)
method in a study based on the CLSI EP09-A3 guideline, “Measurement Procedure Comparison and Bias
Estimation Using Patient Samples”. The samples were analyzed in singlicate over 4 days using 1 instrument.
The range of values on the D-10 Hemoglobin A1c Program was 3.9 to approximately 19.0% (19 to approximately
184 mmol/mol) HbA1c. The results of the method comparison are presented in Figures 1a (NGSP %) and 1b
(IFCC mmol/mol). The D-10 Hemoglobin A1c Program estimated bias compared to the NGSP SRL Method is
presented in Table 4.

D-10 Hemoglobin A1c Program vs NGSP SRL D-10 Hemoglobin A1c Program vs NGSP SRL
%HbA1c Correlation mmol/mol HbA1c Correlation
180
18
160
16
D-10 Hemoglobin A1c Program
D-10 Hemoglobin A1c Program

140
14
120
mmol/mol HbA1c
12
100
% HbA1c

10
80
8 60
n = 128 n = 128
6 y = 0.9701x + 0.1801 40 y = 0.9701x + 1.2658
R2 = 0.9955 R2 = 0.9955
4 20

2 0
2 4 6 8 10 12 14 16 18 0 20 40 60 80 100 120 140 160 180

NGSP SRL mmol/mol HbA1c

NGSP SRL % HbA1c

Figure 1a: Correlation of D-10 Hemoglobin A1c Figure 1b: Correlation of D-10 Hemoglobin A1c
Program vs NGSP SRL Method (NGSP %) Program vs NGSP SRL Method
(IFCC mmol/mol)
% HbA1c Bias (% HbA1c) % Bias
5.0 −0.05 −0.96
6.5 0.00 0.03
8.0 −0.08 −0.98
12.0 −0.10 −0.87
Table 4: D-10 Hemoglobin A1c Program Estimated Bias
Linearity
To demonstrate the linearity of the HbA1c measurement throughout the reportable range, a normal and a diabetic
HbA1c whole blood patient sample were used to prepare dilutions, and the diluted samples were analyzed with
the D-10 Hemoglobin A1c Program. The linearity was assessed following the CLSI EP06-A guideline “Evaluation
of the Linearity of Quantitative Measurement Procedures: A Statistical Approach”. The results of the study
demonstrate HbA1c linearity from 3.9−18.8% (19–182 mmol/mol) within a maximum measured difference of
± 0.1% (or ± 1.0 mmol/mol) in this interval.

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 D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Review all sample chromatograms before reporting results. Refer to the Quick Guide for a summary of the
acceptance criteria and result review guidelines.
Please note these characteristics when reviewing chromatograms:
• A1c peak shape: Sharp and uniform (i.e., NOT broad, shouldered, or tailing).
• Baseline: Starts at 0.02 on Y-axis; stable with no ramping.
The following are flagged with an asterisk (*) in the sample report:
• Total area outside of the acceptable range
• HbA1c result outside of the reportable range
• HbF peak concentration >10%

LB002933revA Instructions For Use 19

D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: AJ5JDA4TFO

Injection date 10/17/2023 03:05 PM
Injection #: 85 D-10 Method: HbA1c
Rack #: H1 Rack position: 10
Bio-Rad v: 5.00-2 S/N: #DJ3B001206

Peak table - ID: AJ5JDA4TFO

Peak [Link] Height Area Area %
A1a 0.14 5068 23845 0.7
A1b 0.29 11370 63028 2.0
F 0.51 2189 11319 0.4
LA1c/CHb-2 0.68 5097 34106 1.1
A1c 0.83 13619 114363 5.2
P3 1.31 31103 167984 5.3
A0 1.41 976833 2771414 87.0
Total Area: 3186058

Concentration: % mmol/mol
A1c 5.2 33

Figure 2: Non-Diabetic (Normal) Result

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 D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: presample

Injection date 10/16/2023 09:44 AM
Injection #: 6 D-10 Method: HbA1c
Rack #: 02 Rack position: 6
Bio-Rad v: 5.00-2 S/N: #DJ3B001206

Peak table - ID: presample

Peak [Link] Height Area Area %
A1a 0.18 4733 20937 0.8
A1b 0.29 12128 78948 3.1
F 0.52 2800 13492 0.5
LA1c/CHb-1 0.63 9654 63197 2.5
A1c 0.81 22934 196109 9.7
P3 1.31 41389 175530 6.9
A0 1.41 718282 1993892 78.4
Total Area: 2542106

Concentration: % mmol/mol
A1c 9.7 83

Figure 3: Diabetic Result

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D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: presample

Injection date 10/12/2023 09:55 AM
Injection #: 17 D-10 Method: HbA1c
Rack #: 01 Rack position: 8
Bio-Rad v: 5.00-2 S/N: #DJ6I044901

Peak table - ID: presample

Peak [Link] Height Area Area %
A1b 0.30 6036 49290 3.0
F 0.52 1309 10460 0.6
LA1c/CHb-1 0.66 2251 8379 0.5
LA1c/CHb-2 0.75 9917 62758 3.8
A1c 0.87 8054 71247 6.0
P3 1.31 47292 153469 9.2
A0 1.41 527529 1306906 78.6
Total Area: 1662510

Concentration: % mmol/mol
A1c 6.0 43

Figure 4: Result with Labile A1c (LA1c/CHb-1) and Carbamylated Hemoglobin (LA1c/CHb-2)

22 Instructions For Use LB002933revA

 D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: RACKH2-6-71-12-10-2023

Injection date 10/12/2023 02:17 PM
Injection #: 71 D-10 Method: HbA1c
Rack #: H2 Rack position: 6
Bio-Rad v: 5.00-2 S/N: #DJ6I044901

Peak table - ID: RACKH2-6-71-12-10-2023

Peak [Link] Height Area Area %
A1a 0.13 7449 14625 0.6
A1b 0.25 24407 102389 3.9
Unknown 0.37 4090 15391 0.6
F 0.58 59182 374553 14.2 *
A1c 0.85 11379 100548 6.2
P3 1.31 31540 137276 5.2
A0 1.40 646991 1888208 71.7
Total Area: 2632990

Concentration: % mmol/mol
A1c 6.2 44

Figure 5: Result with Elevated HbF

NOTE: HbF >10.0%, HbA1c result should not be reported.

LB002933revA Instructions For Use 23

D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: RACK04-5-47-3-10-2023

Injection date 10/03/2023 11:33 AM
Injection #: 47 D-10 Method: HbA1c
Rack #: 04 Rack position: 5
Bio-Rad v: 5.00-2 S/N: #DJ6I044901

Peak table - ID: RACK04-5-47-3-10-2023

Peak [Link] Height Area Area %
A1a 0.13 4334 10630 0.6
A1b 0.29 3475 24577 1.4
F 0.53 1749 9553 0.5
LA1c/CHb-1 0.71 1406 9423 0.5
LA1c/CHb-2 0.77 1619 10234 0.6
A1c 0.88 3935 34195 4.7
P3 1.32 14288 71394 4.0
A0 1.42 353162 899945 50.0
D,S-window 1.65 451804 728742 40.5
Total Area: 1798692

Concentration: % mmol/mol
A1c 4.7 28

Figure 6: Result with Hemoglobin S Trait, or Sickle Cell Trait (AS)

24 Instructions For Use LB002933revA

 D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: RACK04-7-49-3-10-2023

Injection date 10/03/2023 11:39 AM
Injection #: 49 D-10 Method: HbA1c
Rack #: 04 Rack position: 7
Bio-Rad v: 5.00-2 S/N: #DJ6I044901

Peak table - ID: RACK04-7-49-3-10-2023

Peak [Link] Height Area Area %
A1a 0.13 4242 10784 0.7
A1b 0.29 3558 21851 1.4
F 0.53 700 3868 0.3
LA1c/CHb-2 0.72 1353 9354 0.6
A1c 0.88 4562 40467 5.6
P3 1.32 12644 63061 4.1
A0 1.42 331765 870551 57.2
C-window 1.80 238873 500705 32.9
Total Area: 1520642

Concentration: % mmol/mol
A1c 5.6 38

Figure 7: Result with Hemoglobin C Trait (AC)

LB002933revA Instructions For Use 25

D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: presample

Injection date 10/16/2023 09:53 AM
Injection #: 9 D-10 Method: HbA1c
Rack #: 02 Rack position: 9
Bio-Rad v: 5.00-2 S/N: #DJ3B001206

Peak table - ID: presample

Peak [Link] Height Area Area %
A1a 0.13 4667 14626 0.6
A1b 0.29 5610 33456 1.4
Unknown 0.45 963 2405 0.1
F 0.52 2136 12527 0.5
LA1c/CHb-2 0.68 3018 20430 0.9
A1c 0.84 5299 45526 5.0
P3 1.32 17098 84071 3.6
A0 1.42 427415 1136230 48.2
D,S-window 1.62 447382 1010450 42.8
Total Area: 2359722

Concentration: % mmol/mol
A1c 5.0 31

Figure 8: Result with Hemoglobin D Trait (AD)

26 Instructions For Use LB002933revA

 D-10™ Hemoglobin A1c Program

SAMPLE REPORTS
Patient report

Sample ID: presample

Injection date 10/13/2023 09:09 AM
Injection #: 14 D-10 Method: HbA1c
Rack #: 01 Rack position: 2
Bio-Rad v: 5.00-2 S/N: #DJ6I044901

Peak table - ID: presample

Peak [Link] Height Area Area %
A1a 0.13 3435 6550 0.3
A1b 0.29 5055 42338 1.9
F 0.51 1836 15266 0.7
LA1c/CHb-2 0.73 2620 17264 0.8
A1c 0.88 9142 78999 7.1
P3 1.34 41758 147574 6.6
A0 1.42 502081 1244431 55.5
E-window 1.55 238813 691085 30.8
Total Area: 2243507

Concentration: % mmol/mol
A1c 7.1 54

Figure 9: Result with Hemoglobin E Trait (AE)

LB002933revA Instructions For Use 27

D-10™ Hemoglobin A1c Program

TRADEMARK INFORMATION
Bio-Rad, D-10, Liquichek, and Lyphochek are trademarks of Bio-Rad Laboratories, Inc. in certain jurisdictions.
All trademarks used herein are the property of their respective owner.

REFERENCES
1. American Diabetes Association Website. About Diabetes. [Link]
basics/?loc=GlobalNavDB (accessed November 2023).
2. Fowler, M. J. Microvascular and Macrovascular Complications of Diabetes. Clin. Diabetes 2008, 26 (2),
77–82.
3. Whiting, D. R.; Guariguata, L.; Weil, C.; Shaw, J. IDF Diabetes Atlas: Global Estimates of the Prevalence of
Diabetes for 2011 and 2030. Diabetes Res. Clin. Pract. 2011, 94, 311–321.
4. American Diabetes Association. Classification and Diagnosis of Diabetes: Standards of Care in
Diabetes—2023. Diabetes Care 2023, 46 (Suppl. 1), S19–S40.
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. Global Report on Diabetes; WHO Press: Geneva, Switzerland, 2016.
7. Forsham, P. H. Diabetes Mellitus: A Rational Plan for Management. Postgrad. Med. 1982, 71, 139–154.
8. Hollander, P. The Case for Tight Control in Diabetes. Postgrad. Med. 1984, 75, 80–87.
9. Baynes, J. W.; Bunn, H. F.; Goldstein, D.; Harris, M.; Martin, D. B.; Peterson, C.; Winterhalter, K. National
Diabetes Data Group: Report of the Expert Committee on Glucosylated Hemoglobin. Diabetes Care 1984,
7, 602–606.
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LB002933revA Instructions For Use 29

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