05336996001V3.

A1C-3
Tina-quant Hemoglobin A1c Gen.3 - Whole blood and Hemolysate Application
Order information
Analyzer(s) on which kit(s) can be used
05336180 190 Tina-quant Hemoglobin A1c Gen.3 (2 × 100 tests) cobas c 111
Materials required (but not provided):
04528417 190 C.f.a.s. HbA1c (3 × 2 mL) Code 674
05479207 190 PreciControl HbA1c norm (4 × 1 mL) Code 208
05912504 190 PreciControl HbA1c path (4 × 1 mL) Code 209
05007232 190 Hemolyzing Reagent Gen.2 (8 × 6.3 mL) Code 952
11488457 122 HbA1c Hemolyzing Reagent for Tina-quant HbA1c (1000 mL)
11930630 001 Chimneys

English Test principle9,10,11

System information This method uses TTAB (Tetradecyltrimethylammonium bromide) as the
detergent in the hemolyzing reagent to eliminate interference from
Whole Blood DCCT (%): ACN 891 A1W3D and ACN 871 HBW3D leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to
Hemolysate DCCT (%): ACN 861 A1H3D and ACN 841 HBH3D remove labile HbA1c is not necessary.
Whole Blood IFCC (mmol/mol): ACN 264 A1W3I and ACN 265 HBW3I
Hemolysate IFCC (mmol/mol): ACN 251 A1H3I and ACN 231 HBH3I All hemoglobin variants which are glycated at the β‑chain N‑terminus and
which have antibody-recognizable regions identical to that of HbA1c are
Intended use measured by this assay. Consequently, the metabolic state of patients
In vitro test for the quantitative determination of mmol/mol hemoglobin A1c having uremia or the most frequent hemoglobinopathies (HbAS, HbAC,
(IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood and HbAE) can be determined using this assay.12,13
hemolysates prepared from whole blood on the cobas c 111 system. Hemoglobin A1c
HbA1c determinations are useful for monitoring of long-term blood glucose The HbA1c determination is based on the turbidimetric inhibition
control in individuals with diabetes mellitus. Moreover, this test is to be used immunoassay (TINIA) for hemolyzed whole blood.
as an aid in diagnosis of diabetes and identifying patients who may be at
risk for developing diabetes. ▪ Sample and addition of R1 (buffer/antibody):
Summary1,2,3,4,5,6,7,8 Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c
antibody to form soluble antigen-antibody complexes. Since the specific
Hemoglobin (Hb) consists of four protein subunits, each containing a heme HbA1c antibody site is present only once on the HbA1c molecule,
moiety, and is the red-pigmented protein located in the erythrocytes. Its complex formation does not take place.
main function is to transport oxygen and carbon dioxide in blood. Each Hb
molecule is able to bind four oxygen molecules. Hb consists of a variety of ▪ Addition of SR (buffer/polyhapten) and start of reaction:
subfractions and derivatives. Among this heterogeneous group of The polyhaptens react with excess anti-HbA1c antibodies to form an
hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction insoluble antibody-polyhapten complex which can be measured
formed by the attachment of various sugars to the Hb molecule. HbA1c is turbidimetrically.
formed in two steps by the nonenzymatic reaction of glucose with the Hemoglobin
N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first Liberated hemoglobin in the hemolyzed sample is converted to a derivative
step is reversible and yields labile HbA1c. This is rearranged to form stable having a characteristic absorption spectrum which is measured
HbA1c in a second reaction step. bichromatically during the preincubation phase (sample + R1) of the above
In the erythrocytes, the relative amount of HbA converted to stable HbA1c immunological reaction. A separate Hb reagent is consequently not
increases with the average concentration of glucose in the blood. The necessary.
conversion to stable HbA1c is limited by the erythrocyte’s life span of The final result is expressed as mmol/mol HbA1c or percent HbA1c and is
approximately 100 to 120 days. As a result, HbA1c reflects the average calculated from the HbA1c/Hb ratio as follows:
blood glucose level during the preceding 2 to 3 months. HbA1c is thus Protocol 1 (mmol/mol HbA1c acc. to IFCC):
suitable to monitor long-term blood glucose control in individuals with HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
diabetes mellitus. Glucose levels closer to the time of the assay have a Protocol 2 (% HbA1c acc. to DCCT/NGSP):
greater influence on the HbA1c level.1 HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
The approximate relationship between HbA1c and mean blood glucose
values during the preceding 2 to 3 months was analyzed in several studies. Reagents – working solutions
A recent study obtained the following correlation: R1 Antibody reagent
IFCC standardization (recalculated acc. to ref. 8) MESa) buffer: 0.025 mol/L; TRISb) buffer: 0.015 mol/L, pH 6.2;
• Estimated average glucose [mmol/L] = 0.146 × HbA1c (mmol/mol) + 0.834 HbA1c antibody (ovine serum) ≥ 0.5 mg/mL; detergents;
or stabilizers; preservative
• Estimated average glucose [mg/dL] = 2.64 × HbA1c (mmol/mol) + 15.03 SR Polyhapten reagent
Standardization acc. to DCCT/NGSP7 MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c
• Estimated average glucose [mmol/L] = 1.59 × HbA1c (%) - 2.59 polyhapten: ≥ 8 µg/mL; detergents; stabilizers; preservative
or a) MES = 2-morpholinoethane sulfonic acid
• Estimated average glucose [mg/dL] = 28.7 × HbA1c (%) - 46.7 b) TRIS = Tris(hydroxymethyl)-aminomethane
The risk of diabetic complications, such as diabetic nephropathy and Precautions and warnings
retinopathy, increases with poor metabolic control. In accordance with its For in vitro diagnostic use for health care professionals. Exercise the
function as an indicator for the mean blood glucose level, HbA1c predicts normal precautions required for handling all laboratory reagents.
the development of diabetic complications in diabetes patients.3,4
Infectious or microbial waste:
For monitoring of long term glycemic control, testing every 3 to 4 months is Warning: handle waste as potentially biohazardous material. Dispose of
generally sufficient. In certain clinical situations, such as gestational waste according to accepted laboratory instructions and procedures.
diabetes, or after a major change in therapy, it may be useful to measure
HbA1c in 2 to 4 week intervals.6 Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
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This kit contains components classified as follows in accordance with the from various manufacturers may contain differing materials which could
Regulation (EC) No. 1272/2008: affect the test results in some cases. When processing samples in primary
tubes (sample collection systems), follow the instructions of the tube
manufacturer.
See the limitations and interferences section for details about possible
sample interferences.

Warning Stability: 3 days at 15‑25 °C

7 days at 2‑8 °C
H317 May cause an allergic skin reaction. 6 months at (‑15)‑(‑25) °C
Prevention: The recovery of HbA1c ratio values from sedimented samples, especially in
case of poorly controlled diabetic patients, may be slightly elevated. To
P261 Avoid breathing dust/fume/gas/mist/vapours/spray. minimize this effect samples may be gently mixed by inversion prior to
analysis.
P272 Contaminated work clothing should not be allowed out of
Freeze only once. Mix specimen thoroughly after thawing.
the workplace.
Whole blood application
P280 Wear protective gloves. Whole blood samples must be placed on the instrument in uncapped
primary tubes and not in (micro) tubes.
Response:
Hemolysate application
P333 + P313 If skin irritation or rash occurs: Get medical Manual hemolysate preparation:
advice/attention. 1. Allow blood specimen and Hemolyzing Reagent for Tina‑quant HbA1c
(Cat. No. 11488457122) to equilibrate at room temperature before use.
P362 + P364 Take off contaminated clothing and wash it before reuse.
2. Moderately mix the sample immediately prior to pipetting, to ensure
Disposal: homogeneous mixture of erythrocytes. Take care to avoid the formation
of foam.
P501 Dispose of contents/container to an approved waste
disposal plant. 3. Dilute the sample with Hemolyzing Reagent for Tina‑quant HbA1c in the
ratio 1:101 (1+100) using one of the following pipetting schemes.
Product safety labeling follows EU GHS guidance.
Pipette into tubes:
Contact phone: all countries: +49-621-7590
Hemolyzing Reagent for Tina-quant HbA1c: 500 µL
Reagent handling Specimen (patient or control): 5 µL
Ready for use or
Under conditions of extreme humidity, condensation may lead to a dilution Hemolyzing Reagent for Tina-quant HbA1c: 1000 µL
of the reagent that affects the measurements. Hence under environmental Specimen (patient or control): 10 µL
conditions in which temperature and humidity are equal to, or in excess of or
25 °C/80 %, 28 °C/70 %, 30 °C/60 % or 32 °C/55 % a chimney Hemolyzing Reagent for Tina-quant HbA1c: 2000 µL
(Cat. No. 11930630 001) should be used to reduce the condensation rate. Specimen (patient or control): 20 µL
Place a white chimney in R1 and a black chimney in SR. The chimneys can 4. Mix using a vibration mixer or by gentle swirling.
be reused for reagent bottles within the same kit. However, to avoid 5. The hemolysate can be used after the solution has changed color from
contamination of the reagent with detergent or dilution of the reagent with red to brownish-green (approximately 1‑2 min).
water it is not permitted to wash the chimneys before reuse.
Storage and stability Stability of the hemolysate: 4 hours at 15‑25 °C
24 hours at 2‑8 °C
Reagent 6 months at (‑15)‑(‑25) °C
Shelf life at 2‑8 °C: See expiration date Materials provided
on reagent
See “Reagents – working solutions” section for reagents.
On-board in use and refrigerated on the analyzer: 4 weeks
Materials required (but not provided)
Hemolyzing Reagent Gen.2, Cat. No. 05007232190 (concentrated) See “Order information” section
Hemolyzing Reagent Gen.2, Cat. No. 05007232190 (concentrated for
Shelf life at 2‑8 °C: See expiration date whole blood testing). This hemolyzing reagent must be on-board the
on vial label analyzer for calibration measurements of both methods.
On-board in use and refrigerated on the analyzer: 5 days Hemolyzing Reagent for Tina‑quant HbA1c, Cat. No. 11488457122 (for
manual dilution). Use this reagent to prepare hemolysate. See above for
When storing at temperatures under 3 °C, the reagent may become cloudy. sample pretreatment procedure.
This has no effect on the function of the reagent and is reversible at higher
temperatures. It is therefore recommended to equilibrate the reagent at General laboratory equipment
room temperature for approximately 10 minutes and mix thoroughly before Assay
use.
For optimum performance of the assay follow the directions given in this
Specimen collection and preparation document for the analyzer concerned. Refer to the appropriate operator’s
For specimen collection and preparation only use suitable tubes or manual for analyzer‑specific assay instructions.
collection containers. The performance of applications not validated by Roche is not warranted
Only the specimens listed below were tested and found acceptable. and must be defined by the user.
Anticoagulated venous or capillary blood or hemolysate Whole Blood applications for Hb and HbA1c
The only acceptable anticoagulants are Li‑Heparin, K2‑EDTA, K3‑EDTA,
Fluoride/Na2‑EDTA, Na‑Heparin and Fluoride/potassium oxalate. cobas c 111 test definition Hb (HBW3I or HBW3D)
The sample types listed were tested with a selection of sample collection Measuring mode Absorbance
tubes that were commercially available at the time of testing, i.e. not all
available tubes of all manufacturers were tested. Sample collection systems Abs. calculation mode Endpoint

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Reaction direction Increase Pipetting parameters

Wavelength A/B 378/659 nm HbA1c Diluent (H2O)
Calc. first/last 6/16 R1 120 µL
Unit mmol/L Sample 6 µL 0 µL
Reaction mode R1-S SR 24 µL 0 µL
Pipetting parameters Total volume 150 µL
Hb Diluent (H2O) Calibration
R1 120 µL Hb
Sample 6 µL 0 µL Calibrator C.f.a.s. HbA1c
Total volume 126 µL HbA1c
cobas c 111 test definition HbA1c (A1W3I or A1W3D) Calibrator C.f.a.s. HbA1c
Measuring mode Absorbance Calibration dilution ratio 1:1, 1:1.5, 1:2.1, 1:3, 1:6, 1:15,
Abs. calculation mode Endpoint performed automatically by the
instrument
Reaction direction Increase
Calibrator diluent Hemolyzing Reagent Gen.2
Wavelength A/B 340/659 nm
Calibration frequency Each lot, every 14 days, and as required
Calc. first/last 16/34
following quality control procedures
Unit mmol/L
Calibration interval may be extended based on acceptable verification of
Reaction mode R1-S-SR calibration by the laboratory.
Traceability: This method has been standardized against the approved
Pipetting parameters IFCC reference method for the measurement of HbA1c in human blood14,15
HbA1c Diluent (H2O) and can be transferred to results traceable to DCCT/NGSP by calculation.
Note
R1 120 µL
Enter the assigned lot-specific and application-specific value of the
Sample 6 µL 0 µL calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only.
SR 24 µL 0 µL The cobas c 111 Hemolyzing Reagent Gen.2 (Cat. No. 05007232 190)
needs to be available on the analyzer. Otherwise the calibration cannot be
Total volume 150 µL performed.
Hemolysate applications for Hb and HbA1c Quality control
cobas c 111 test definition Hb (HBH3I or HBH3D) For quality control, use control materials as listed in the “Order information”
section. In addition, other suitable control material can be used.
Measuring mode Absorbance The control intervals and limits should be adapted to each laboratory’s
Abs. calculation mode Endpoint individual requirements. Values obtained should fall within the defined
limits. Each laboratory should establish corrective measures to be taken if
Reaction direction Increase values fall outside the defined limits.
Wavelength A/B 378/659 nm Follow the applicable government regulations and local guidelines for
quality control.
Calc. first/last 6/16
Note
Unit mmol/L HbA1c controls carry a declaration for HbA1c ratio only. No declarations for
Reaction mode R1-S Hb and HbA1c concentrations are provided. HbA1c controls must be
included in the cobas c 111 systems Quality Control Program. For the
Pipetting parameters hemolysate test the HbA1c controls must be hemolysed manually before
using them for the Quality Control Program. Assigned target values in
Hb Diluent (H2O) mmol/mol HbA1c according to IFCC are not included in the control
R1 120 µL barcodes and must be entered by the customer manually.
Sample 6 µL 0 µL Calculation
The cobas c 111 analyzer automatically calculates the ratio between A1c
Total volume 126 µL and Hb of each sample.
cobas c 111 test definition HbA1c (A1H3I or A1H3D) Limitations - interference12,13,16,17,18,19,20,21,22,23
1. For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c
Measuring mode Absorbance values (DCCT/NGSP) should be used in conjunction with information
Abs. calculation mode Endpoint from other diagnostic procedures and clinical evaluations.
Reaction direction Increase 2. The test is designed only for accurate and precise measurement of
mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
Wavelength A/B 340/659 nm The individual results for total Hb and HbA1c concentration should not
Calc. first/last 16/34 be reported.
Unit mmol/L
Reaction mode R1-S-SR

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3. As a matter of principle, care must be taken when interpreting any In rare cases of ">test rng" flags that might occur with the whole blood
HbA1c result from patients with Hb variants. Abnormal hemoglobins application remix the whole blood sample and repeat the analysis with the
might affect the half life of the red cells or the in vivo glycation rates. In same settings.
these cases even analytically correct results do not reflect the same Lower limits of measurement
level of glycemic control that would be expected in patients with normal
hemoglobin.21 Limit of Blank and Limit of Detection
Whenever it is suspected that the presence of an Hb variant (e.g. HbSS,
HbCC, or HbSC) affects the correlation between the HbA1c value and Hb: Limit of Blank = 0.31 mmol/L (0.50 g/dL)
glycemic control HbA1c must not be used for the diagnosis of diabetes Limit of Detection = 0.62 mmol/L (1.00 g/dL)
mellitus.
HbA1c: Limit of Blank = 0.12 mmol/L (0.19 g/dL)
4. Any cause of shortened erythrocyte survival will reduce exposure of
erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c Limit of Detection = 0.18 mmol/L (0.29 g/dL)
values (IFCC) and % HbA1c values (DCCT/NGSP), even though the The Limit of Blank and Limit of Detection were determined in accordance
time-averaged blood glucose level may be elevated. Causes of with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A
shortened erythrocyte lifetime might be hemolytic anemia or other requirements.
hemolytic diseases, homozygous sickle cell trait, pregnancy, recent
significant or chronic blood loss, etc. Caution should be used when The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of
interpreting the HbA1c results from patients with these conditions. analyte‑free samples over several independent series. The Limit of Blank
HbA1c must not be used for the diagnosis of diabetes mellitus in the corresponds to the concentration below which analyte‑free samples are
presence of such conditions. found with a probability of 95 %.
5. Glycated HbF is not detected by the assay as it does not contain the The Limit of Detection is determined based on the Limit of Blank and the
glycated β-chain that characterizes HbA1c. However, HbF is measured standard deviation of low concentration samples.
in the total Hb assay and as a consequence, specimens containing high The Limit of Detection corresponds to the lowest analyte concentration
amounts of HbF (≥ 10 %) may result in lower than expected mmol/mol which can be detected (value above the Limit of Blank with a probability of
HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).13,23 95 %).
6. mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) Expected values
are not suitable for diagnosis of gestational diabetes.24 Protocol 1 (mmol/mol HbA1c acc. to IFCC): 29‑42 mmol/mol HbA1c27
7. In very rare cases of rapidly evolving type 1 diabetes the increase of Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.8‑5.9 % HbA1c27
HbA1c values might be delayed compared to the acute increase in
glucose concentrations. In these conditions diabetes mellitus must be This reference range was obtained by measuring 482 well-characterized
diagnosed based on plasma glucose concentrations and/or the typical healthy individuals without diabetes mellitus. HbA1c levels higher than the
clinical symptoms.24 upper end of this reference range are an indication of hyperglycemia during
the preceding 2 to 3 months or longer. According to the recommendations
Criterion: Recovery within ± 10 % of initial value. of the American Diabetes Association values above 48 mmol/mol HbA1c
Icterus: No significant interference up to a conjugated and unconjugated (IFCC) or 6.5 % HbA1c (DCCT/NGSP) are suitable for the diagnosis of
bilirubin concentration of 1026 μmol/L or 60 mg/dL. diabetes mellitus.24,28 Patients with HbA1c values in the range of
Lipemia: No significant interference up to an Intralipid concentration of 39‑46 mmol/mol HbA1c (IFCC) or 5.7‑6.4 % HbA1c (DCCT/NGSP) may be
600 mg/dL. There is poor correlation between the triglycerides at a risk of developing diabetes.24,28
concentration and turbidity. HbA1c levels may reach 195 mmol/mol HbA1c (IFCC) or 20 % HbA1c
Glycemia: No significant interference up to a glucose level of 55.5 mmol/L (DCCT/NGSP) and more in poorly controlled diabetes. Therapeutic action is
(1000 mg/dL). A fasting sample is not required. suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c
(DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol
Rheumatoid factors: No significant interference up to a rheumatoid factor HbA1c (IFCC) or 7 % HbA1c (DCCT/NGSP) meet the goal of the American
level of 750 IU/mL. Diabetes Association.18,19
Drugs: No interference was found at therapeutic concentrations using HbA1c levels below the established reference range may indicate recent
common drug panels.25,26 episodes of hypoglycemia, the presence of Hb variants, or shortened
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated lifetime of erythrocytes.
hemoglobin, carbamylated hemoglobin, glycated albumin and labile HbA1c Each laboratory should investigate the transferability of the expected values
were found for the anti-HbA1c antibodies used in this kit. to its own patient population and if necessary determine its own reference
For diagnostic purposes, the results should always be assessed in ranges.
conjunction with the patient’s medical history, clinical examination and other
findings. Specific performance data
Representative performance data on the cobas c 111 analyzer are given
ACTION REQUIRED below. Results obtained in individual laboratories may differ.
Special Wash Programming: The use of special wash steps is mandatory
when certain test combinations are run together on the cobas c 111 Precision
analyzer. For information about test combinations requiring special wash Repeatability and intermediate precision were determined using human
steps, please refer to the latest version of the carry-over evasion list found samples and controls in accordance with the CLSI (Clinical and Laboratory
with the CLEAN Method Sheet and the operator’s manual for further Standards Institute) EP5 requirements (2 aliquots per run, 2 runs per day,
instructions. 21 days). The following results were obtained (data based on DCCT/NGSP
Where required, special wash/carry-over evasion programming must values):
be implemented prior to reporting results with this test.
Whole Blood application
Limits and ranges
Measuring range Repeatability Mean SD CV
Hb: 2.48‑24.8 mmol/L (4‑40 g/dL) % HbA1c % HbA1c %
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)* PreciControl HbA1c norm 5.63 0.04 0.7
*The measuring range for HbA1c lies between 0.186 mmol/L and the PreciControl HbA1c path 10.7 0.06 0.6
concentration of the highest standard. The test range stated above is based
on a typical calibrator value of 1.61 mmol/L. Human sample 1 5.21 0.04 0.7
This corresponds to a measuring range of 23‑196 mmol/mol HbA1c (IFCC) Human sample 2 6.22 0.04 0.7
and 4.2‑20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin
concentration of 8.2 mmol/L (13.2 g/dL). Human sample 3 7.34 0.05 0.7

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Repeatability Mean SD CV Hemolysate application:

% HbA1c % HbA1c % % HbA1c (DCCT/NGSP) values for human blood samples obtained on a
cobas c 111 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent
Human sample 4 8.44 0.05 0.6 with the hemolysate application (y) were compared to those determined
Human sample 5 11.0 0.06 0.5 using the same reagent with the hemolysate application on a
COBAS INTEGRA 400 plus analyzer (x).
Intermediate precision Mean SD CV
Sample size (n) = 57
% HbA1c % HbA1c %
Mean difference: 0.19 % HbA1c
PreciControl HbA1c norm 5.63 0.08 1.5
Lower 95 % confidence interval of differences: −0.16 % HbA1c
PreciControl HbA1c path 10.7 0.10 0.9
Upper 95 % confidence interval of differences: 0.54 % HbA1c
Human sample 1 5.21 0.07 1.4
The sample concentrations were between 4.58 % and
Human sample 2 6.22 0.07 1.1 11.8 % HbA1c (DCCT/NGSP values).
Human sample 3 7.34 0.09 1.2 Analytical specificity
Human sample 4 8.44 0.09 1.1
Hb derivatives Labile HbA1c (pre-HbA1c), acetylated Hb, and
Human sample 5 11.0 0.10 0.9 carbamylated Hb do not affect the assay result.
Hemolysate application Hb variants Specimens containing high amounts of HbF (> 10 %)
Repeatability Mean SD CV may yield lower than expected HbA1c results.
% HbA1c % HbA1c % Please note:
PreciControl HbA1c norm 5.76 0.06 1.0 According to the consensus statement of the American Diabetes
Association (ADA), the European Association for the Study of Diabetes
PreciControl HbA1c path 10.5 0.06 0.6 (EASD), the International Federation of Clinical Chemistry and Laboratory
Medicine (IFCC) and the International Diabetes Federation (IDF) HbA1c
Human sample 1 4.99 0.07 1.5 results should be reported in parallel, both in mmol/mol HbA1c (IFCC) and
Human sample 2 6.42 0.08 1.3 % HbA1c (DCCT/NGSP) values.29 In addition an HbA1c derived estimated
average glucose concentration can be reported which can be calculated
Human sample 3 10.2 0.07 0.7 according to the equations given in the summary section of this method
Human sample 4 7.44 0.07 0.9 sheet. Former % HbA1c (IFCC) values must not be used due to the risk of
mix up / misinterpretation with the % HbA1c (DCCT/NGSP) values.
Human sample 5 7.69 0.05 0.7
To ensure parallel reporting of results in both, IFCC units (mmol/mol) and
DCCT/NGSP units (%) on the cobas c 111 analyzer without having to
Intermediate precision Mean SD CV request the assay twice, an additional ratio must be defined on the
% HbA1c % HbA1c % analyzer. Please contact your local Roche Diagnostics support if you
PreciControl HbA1c norm 5.76 0.12 2.1 require any assistance with programming this ratio; separate customer
information is available on request.
PreciControl HbA1c path 10.5 0.19 1.8 Analyzer software version 3.0 and later is a prerequisite for reporting the
Human sample 1 4.99 0.11 2.2 new IFCC units (mmol/mol).
Human sample 2 6.42 0.16 2.5 References
Human sample 3 10.2 0.18 1.8 1 Goldstein DE, Little RR, Lorenz RA, et al. Tests of glycemia in
diabetes. Diabetes Care 1995;18:896-909.
Human sample 4 7.44 0.14 1.9 2 Goldstein DE, Little RR. More than you ever wanted to know (but need
Human sample 5 7.69 0.13 1.6 to know) about glycohemoglobin testing. Diabetes Care
1994;17:938-939.
Method comparison 3 The Diabetes Control and Complications Trial Research Group. The
Evaluation of method comparison data is according to former NGSP effect of intensive treatment of diabetes on the development and
certification criteria. The mean difference between the two methods and the progression of long-term complications in insulin-dependent diabetes
95 % confidence intervals of the differences in the range from 4‑10 % mellitus. N Engl J Med 1993;329:977-986.
(DCCT/NGSP) are given. 95 % of the differences between the values
obtained for individual samples with both methods fall within the range 4 UK Prospective Diabetes Study (UKPDS) group. Intensive blood
defined by the lower and upper 95 % confidence intervals of the glucose control with sulfonylureas or insulin compared with
differences. conventional treatment and risk of complications in patients with type 2
diabetes (UKPDS 33). Lancet 1998;352:837-853.
Whole Blood application:
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a 5 Finke A, Kobold U, Hoelzel W, et al. Preparation of a candidate primary
cobas c 111 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent reference material for the international standardization of HbA1c
with the whole blood application (y) were compared to those determined determinations. Clin Chem Lab Med 1998;36(5):299-308.
using the same reagent with the whole blood application on a 6 Goldstein DE, Little RR, Wiedmeyer HM, et al. Glycated hemoglobin:
COBAS INTEGRA 400 plus analyzer (x). methodologies and clinical applications. Clin Chem 1986;32:B64-B70.
Sample size (n) = 57 7 Nathan DM, Kuenen J, Borg R, et al. Translating the A1C assay into
estimated average glucose values. Diabetes Care 2008;31:1473-1478.
Mean difference: 0.22 % HbA1c 8 Bunn HF, Gabbay KH, Gallop PM. The glycosylation of hemoglobin:
Lower 95 % confidence interval of differences: −0.22 % HbA1c relevance to diabetes mellitus. Science 1978;200:21-27.
Upper 95 % confidence interval of differences: 0.66 % HbA1c 9 Zander R, Lang W, Wolf HU. Alkaline haematin D-575, a new tool for
the determination of haemoglobin as an alternative to the
The sample concentrations were between 4.86 % and cyanhaemiglobin method. I. Description of the method. Clin Chim Acta
12.2 % HbA1c (DCCT/NGSP values). 1984;136:83-93.

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10 Wolf HU, Lang W, Zander R. Alkaline haematin D-575, a new tool for Any serious incident that has occurred in relation to the device shall be
the determination of haemoglobin as an alternative to the reported to the manufacturer and the competent authority of the Member
cyanhaemiglobin method. II. Standardization of the method using pure State in which the user and/or the patient is established.
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Roche Diagnostics uses the following symbols and signs in addition to
12 Frank EL, Moulton L, Little RR, et al. Effects of hemoglobin C and S those listed in the ISO 15223‑1 standard (for USA: see dialog.roche.com for
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2000;46(6):864-867.
13 Chang J, Hoke C, Ettinger B, et al. Evaluation and Interference Study Contents of kit
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Volume after reconstitution or mixing
14 Kobold U, Jeppsson JO, Duelffer T, et al. Candidate reference methods
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15 Jeppsson JO, Kobold U, Finke A, et al. Approved IFCC reference COBAS, COBAS C, COBAS INTEGRA, TINA-QUANT and PRECICONTROL are trademarks of Roche.
method for the measurement of HbA1c in human blood. Clin Chem Lab All other product names and trademarks are the property of their respective owners.
Med 2002;40:78-89. Additions, deletions or changes are indicated by a change bar in the margin.
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Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
17 Weykamp CW, Penders TJ, Muskiet FAJ, et al. Influence of www.roche.com
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28 Diagnosis and Classification of Diabetes Mellitus. Diabetes Care
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29 Consensus statement on the worldwide standardization of the
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A point (period/stop) is always used in this Method Sheet as the decimal
separator to mark the border between the integral and the fractional parts of
a decimal numeral. Separators for thousands are not used.

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