04528123190V3

A1C-2
Tina-quant Hemoglobin A1c Gen.2
• Indicates cobas c systems on which reagents can be used
Order information Roche/Hitachi cobas c systems
Tina-quant Hemoglobin A1c Gen.2 cobas c 501
150 tests Cat. No. 04528123 190 System-ID 07 6850 2 •
C.f.a.s. HbA1c (3 x 2 mL) Cat. No. 04528417 190 Code 674
HbA1c Control N (4 × 0.5 mL) Cat. No. 20764833 322 Code 357
HbA1c Control P (4 x 0.5 mL) Cat. No. 20764841 322 Code 358
Hemolyzing Reagent Gen.2 (2 x 22.5 mL) Cat. No. 04528182 190 System-ID 07 6873 1
HbA1c Hemolyzing Reagent for Tina-quant HbA1c
Cat. No. 11488457 122 For Hemolysate Application only
(1000 mL)
English Test principle10,11,12
This method uses TTAB* as the detergent in the hemolyzing reagent to
System information
eliminate interference from leukocytes (TTAB does not lyse leukocytes).
Whole Blood Application - Standardized according to IFCC transferable Sample pretreatment to remove labile HbA1c is not necessary.
to DCCT/NGSP All hemoglobin variants which are glycated at the β-chain N-terminus and
HB-W2: ACN 870 Hemoglobin (Hb) which have antibody-recognizable regions identical to that of HbA1c are
A1-W2: ACN 880 Hemoglobin A1c (HbA1c) measured by this assay. Consequently, the metabolic state of patients
RWI2: ACN 890 % Ratio having uremia or the most frequent hemoglobinopathies (HbAS, HbAC,
A1CD2: ACN 952 Hemolyzing reagent HbAE) can be determined using this assay.13,14
*TTAB = Tetradecyltrimethylammonium bromide
Hemolysate Application - Standardized according to IFCC transferable Hemoglobin A1c
to DCCT/NGSP The HbA1c determination is based on the turbidimetric inhibition
immunoassay (TINIA) for hemolyzed whole blood.
HB-H2 ACN 840 Hemoglobin (Hb)
• Sample and addition of R1 (buffer/antibody):
A1–H2 ACN 850 Hemoglobin A1c (HbA1c)
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c
RHI2 ACN 860 % Ratio
antibody to form soluble antigen-antibody complexes. Since the
A1CD2 ACN 952 Hemolyzing reagent specific HbA1c antibody site is present only once on the HbA1c
Intended use molecule, complex formation does not take place.
In vitro test for the quantitative determination of percent hemoglobin A1c • Addition of R2 (buffer/polyhapten) and start of reaction:
[HbA1c (%)] in whole blood or in hemolysate on Roche/Hitachi cobas c systems. The polyhaptens react with excess anti-HbA1c antibodies to
form an insoluble antibody-polyhapten complex which can
Summary1,2,3,4,5,6,7,8,9 be measured turbidimetrically.
Hemoglobin (Hb) consists of four protein subunits, each containing a heme Hemoglobin
moiety, and is the red-pigmented protein located in the erythrocytes. Its main Liberated hemoglobin in the hemolyzed sample is converted to a derivative
function is to transport oxygen and carbon dioxide in blood. Each Hb molecule having a characteristic absorption spectrum which is measured bichromatically
is able to bind four oxygen molecules. Hb consists of a variety of subfractions during the preincubation phase (sample + R1) of the above immunological
and derivatives. Among this heterogeneous group of hemoglobins HbA1c is reaction. A separate Hb reagent is consequently not necessary.
one of the glycated hemoglobins, a subfraction formed by the attachment The final result is expressed as percent HbA1c and is calculated
of various sugars to the Hb molecule. HbA1c is formed in two steps by the from the HbA1c/Hb ratio as follows:
nonenzymatic reaction of glucose with the N-terminal amino group of the
β-chain of normal adult Hb (HbA). The first step is reversible and yields labile Protocol 1 (acc. to IFCC):
HbA1c. This is rearranged to form stable HbA1c in a second reaction step. HbA1c (%) = (HbA1c/Hb) × 100
In the erythrocytes, the relative amount of HbA converted to stable Protocol 2 (acc. to DCCT/NGSP):
HbA1c increases with the average concentration of glucose in the HbA1c (%) = (HbA1c/Hb) × 87.6 + 2.27
blood. The conversion to stable HbA1c is limited by the erythrocyte’s Reagents – working solutions
life span of approximately 100 to 120 days. As a result, HbA1c reflects
R1 Antibody reagent
the average blood glucose level during the preceding 2 to 3 months.
MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c
HbA1c is thus suitable to monitor long-term blood glucose control in
antibody (ovine serum): ≥ 0.5 mg/mL; stabilizers; preservatives (liquid)
individuals with diabetes mellitus. Glucose levels closer to the time of
the assay have a greater influence on the HbA1c level.1 R2 Polyhapten reagent
The approximate relationship between HbA1c and mean blood glucose MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c
values during the preceding 2 to 3 months was analyzed in several polyhapten: ≥ 8 µg/mL; stabilizers; preservatives (liquid)
studies. A recent study obtained the following correlation:
Precautions and warnings
IFCC standardization (recalculated acc. to ref. 8) For in vitro diagnostic use.
• Mean plasma glucose [mmol/L] = 1.73 x HbA1c (%) + 0.20 or Exercise the normal precautions required for handling all laboratory reagents.
• Mean plasma glucose [mg/dL] = 31.2 x HbA1c (%) + 3.51 Safety data sheet available for professional user on request.
Standardization acc. to DCCT/NGSP8 Disposal of all waste material should be in accordance with local guidelines.
• Mean plasma glucose [mmol/L] = 1.98 × HbA1c (%) - 4.29 or
Reagent handling
• Mean plasma glucose [mg/dL] = 35.6 × HbA1c (%) - 77.3 Ready for use.
The risk of diabetic complications, such as diabetic nephropathy and
retinopathy, increases with poor metabolic control. In accordance with its
function as an indicator for the mean blood glucose level, HbA1c predicts
the development of diabetic complications in diabetes patients.3,5
For routine clinical use, testing every 3 to 4 months is generally sufficient. In
certain clinical situations, such as gestational diabetes, or after a major change
in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.7
2007-03, V 3 English 1/6 cobas c systems
A1C-2
Tina-quant Hemoglobin A1c Gen.2
Storage and stability Whole Blood application for Hb (HB-W2) and HbA1c (A1-W2)
A1C-2 cobas c 501 test definition Hb (HB-W2)
Shelf life at 2-8°C: See expiration date on Assay type 1 Point
cobas c pack label. Reaction time / Assay points 10 / 34
On-board in use and refrigerated on the analyzer: 4 weeks Wavelength (sub/main) 660/376 nm
Reaction direction Increase
Hemolyzing reagent
Unit g/dL
Shelf life at 2-8°C: See expiration date on
pack label Reagent pipetting Diluent (H2O)
When storing at temperatures under 3°C, the reagent may become cloudy. R1 120 µL -
This has no effect on the function of the reagent and is reversible at higher R3 24 µL -
temperatures. It is therefore recommended to equilibrate the reagent at room
temperature for approximately 10 minutes and mix thoroughly before use. Sample volumes Sample Sample dilution
Sample Diluent
On-board in use and refrigerated on the analyzer: 4 weeks (Hemolyzing
Specimen collection and preparation reagent)
For specimen collection and preparation, only use suitable
Normal 5 µL 2 µL 180 µL
tubes or collection containers.
Decreased 5 µL 2 µL 180 µL
Only the specimens listed below were tested and found acceptable.
Increased 5 µL 2 µL 180 µL
Venous or capillary blood with anticoagulant.
The only acceptable anticoagulants are Li-heparin, K2-EDTA and K3-EDTA.
The sample types listed were tested with a selection of sample collection tubes cobas c 501 test definition HbA1c (A1-W2)
that were commercially available at the time of testing, i.e. not all available Assay type 2 Point End
tubes of all manufacturers were tested. Sample collection systems from Reaction time / Assay points 10 / 34-70
various manufacturers may contain differing materials which could affect Wavelength (sub/main) 660/340 nm
the test results in some cases. When processing samples in primary tubes Reaction direction Increase
(sample collection systems), follow the instructions of the tube manufacturer. Unit g/dL
Stability:15 3 days at 15-25°C Reagent pipetting Diluent (H2O)
7 days at 2-8°C R1 120 µL -
6 months at (-15)-(-25)°C R3 24 µL -

Freeze only once. Mix specimen thoroughly before use. Sample volumes Sample Sample dilution
Hemolysate preparation for Hemolysate Application Sample Diluent
(Hemolyzing
1. Allow blood specimen and Hemolyzing Reagent for Tina-quant HbA1c
reagent)
to equilibrate at room temperature before use.
2. Moderately mix the sample immediately prior to pipetting to Normal 5 µL 2 µL 180 µL
ensure a homogeneous mixture of erythrocytes. Take care Decreased 5 µL 2 µL 180 µL
to avoid the formation of foam. Increased 5 µL 2 µL 180 µL
3. Dilute the sample with Hemolyzing Reagent for Tina-quant HbA1c
(Cat. No. 11488457) in the ratio 1:101 (1+100) using one of Important: To improve the fit of the nonlinear HbA1c calibration curve, a
the following pipetting schemes. constant and lot independent offset of 0.6 g/dL was added to all calibrator
Pipette into tubes: values. This offset is already included in the assigned HbA1c calibrator
target values for cobas c analyzers and needs to be subtracted from
HbA1c Hemolyzing Reagent 500 µL 1000 µL 2000 µL the analyzer’s results. Enter the instrument factor of the absolute HbA1c
for Tina-quant HbA1c (A1-W2) assay on Roche cobas c analyzers as follows:
Specimen (patient or control) 5 µL 10 µL 20 µL Calibration => Status Screen => Instrument Factor => Instrument Factor
4. Mix using a vibration mixer or by gentle swirling. Window => HbA1c (A1-W2) => a = 1.0; b = – 0.6 => update => okay
5. The hemolysate can be used after the solution has changed color Ratio definition for HbA1c (%) calculation
from red to brownish-green (approx. 1-2 min).
Protocol 1 (acc. to IFCC):
Stability of the hemolysate:15 4 hours at 15-25°C
24 hours at 2-8°C Abbreviated ratio name RWI2 (890)
6 months at (-15)-(-25)°C Equation (A1-W2/HB-W2) × 100
Unit %

Materials provided Protocol 2 (acc. to DCCT/NGSP):

See “Reagents - working solutions” section for reagents.
Abbreviated ratio name RWD2
Materials required (but not provided) Equation (A1-W2/HB-W2) × 87.6 + 2.27
See “Order information” section. Unit %
Distilled water
General laboratory equipment Protocol 1 is already implemented in the application (ACN 890). Percent
HbA1c according to Protocol 2 (DCCT/NGSP) must be manually calculated
Assay according to the above equation. If requested the formula (ACN 890) can
For optimum performance of the assay, follow the directions given in be modified by using the Administrator Level/EDIT Button.
this document for the analyzer concerned. Refer to the appropriate The ratio for HbA1c (%) will be automatically calculated after
operator manual for analyzer-specific assay instructions. result output of both tests.
The performance of applications not validated by Roche is not It is recommended to report HbA1c values to one decimal place, which
warranted and must be defined by the user. can be entered in the editable field “expected values”.
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04528123190V3

A1C-2
Tina-quant Hemoglobin A1c Gen.2
Hemolysate Application for Hb (HB-H2 and HbA1c (A1–H2) Calibration for Whole Blood and Hemolysate Application
cobas c 501 test definition Hb (HB-H2) Hb
Assay type 1 Point Calibrators S1-S2: C.f.a.s. HbA1c
Reaction time/Assay points: 10 / 34 Calibration mode Linear
Wavelength (sub/main) 660/376 nm
Reaction direction Increase HbA1c
Unit g/dL Calibrators S1-S6: C.f.a.s. HbA1c
Reagent pipetting Diluent (H2O) Calibration mode RCM
R1 120 µL - Calibration frequency Hb and HbA1c: full calibration is recommended
R3 24 µL - - after 29 days during shelf life
- after reagent lot change
Sample volumes Sample Sample dilution - and as required following quality control
Sample Diluent procedures
Always calibrate both assays (Hb and HbA1c)
in parallel. Automatic calibration at QC failure
Normal 5 µL - - should be deactivated.
Decreased 5 µL - -
Increased 5 µL - - Traceability: This method has been standardized against the approved IFCC
reference method for the measurement of HbA1c in human blood16,17 and
cobas c 501 test definition HbA1c (A1-H2) can be transferred to results traceable to DCCT/NGSP by calculation.
Assay type 2 Point End Note for Whole Blood and Hemolysate Application
Reaction time/Assay points: 10 / 34 -70 Enter the assigned lot-specific and application-specific value of the calibrator.
Wavelength (sub/main) 660/340 nm Use the appropriate C.f.a.s. HbA1c calibrator only.
Reaction direction Increase The cobas c Hemolyzing Reagent Gen.2 pack, 2 x 22.5 mL,
Unit g/dL Cat. No. 04528182, needs to be available on the analyzer.
Reagent pipetting Diluent (H2O) Otherwise the calibration cannot be performed.
R1 120 µL -
- Quality control for Whole Blood and Hemolysate Application
R3 24 µL
For quality control, use control materials as listed in the
Sample volumes Sample Sample dilution “Order information” section.
Sample Diluent Other suitable control material can be used in addition.
The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined
limits. Each laboratory should establish corrective measures to be
Normal 5 µL - -
taken if values fall outside the limits.
Decreased 5 µL - -
Increased 5 µL - - Calculation for Whole Blood and Hemolysate Application
Hb, HbA1c
Important: To improve the fit of the nonlinear HbA1c calibration curve a Roche/Hitachi cobas c systems automatically calculate the analyte
constant and lot independent offset of 0.6 g/dL was added to all calibrator concentration of each sample.
values. This offset is already included in the assigned HbA1c calibrator
target values for cobas c and finally needs to be subtracted from the HbA1c (%)
analyzer’s results. Enter the instrument factor of the absolute HbA1c For calculation of the percent HbA1c value, refer to the Test principle and
(A1-H2) assay on Roche cobas c analyzers as follows: Ratio definition for HbA1c (%) calculation sections in this method sheet.
Calibration => Status Screen => Instrument Factor => Instrument Factor Limitations - interference for Whole Blood and Hemolysate
Window => HbA1c (A1-H2) => a= 1.0; b= – 0.6 => update => okay Application13,14,18,19,20,21,22,23,24,25
Ratio definition for HbA1c (%) calculation 1. For diagnostic purposes, HbA1c (%) values should be used in conjunction
Protocol 1 (acc. to IFCC): with information from other diagnostic procedures and clinical evaluations.
2. The test is designed only for accurate and precise measurement
Abbreviated ratio name RHI2 (860) of HbA1c (%). The individual results for total Hb and HbA1c
Equation (A1-H2/HB-H2) × 100 concentration should not be reported.
Unit % 3. The test is not intended for the diagnosis of diabetes mellitus or
for judging day-to-day glucose control and should not be used to
Protocol 2 (acc. to DCCT/NGSP): replace daily home testing of urine or blood glucose.
Abbreviated ratio name RHD2 4. As a matter of principle, care must be taken when interpreting any HbA1c
Equation (A1-H2/HB-H2) × 87.6 + 2.27 result from patients with Hb variants. Abnormal hemoglobins might affect
the half life of the red cells or the in vivo glycation rates. In these cases
Unit %
even analytically correct results do not reflect the same level of glycemic
Protocol 1 is already implemented in the application (ACN 860). Percent control that would be expected in patients with normal hemoglobin.23
HbA1c according to Protocol 2 (DCCT/NGSP) must be manually calculated 5. Any cause of shortened erythrocyte survival will reduce exposure of
according to the above equation. If requested the formula (ACN 890) can erythrocytes to glucose with a consequent decrease in HbA1c (%) values,
be modified by using the Administrator Level/EDIT Button. even though the time-averaged blood glucose level may be elevated.
The ratio for HbA1c (%) will be automatically calculated after Causes of shortened erythrocyte lifetime might be hemolytic anemia
result output of both tests. or other hemolytic diseases, homozygous sickle cell trait, pregnancy,
It is recommended to report HbA1c values to one decimal place, which recent significant or chronic blood loss, etc. Caution should be used when
can be entered in the editable field “expected values.” interpreting the HbA1c results from patients with these conditions.
2007-03, V 3 English 3/6 cobas c systems
A1C-2
Tina-quant Hemoglobin A1c Gen.2
6. Glycated HbF is not detected by the assay as it does not contain Specific performance data for Whole Blood Application
the glycated ß-chain that characterizes HbA1c. However, HbF Representative performance data on the analyzers are given below.
is measured in the Total Hb assay and as a consequence, Results obtained in individual laboratories may differ.
specimens containing high amounts of HbF (>10%) may result
in lower than expected HbA1c values.14,25 Precision
Reproducibility was determined using human samples and controls in an
Criterion: Recovery within ±10% of initial value at a decision
internal protocol (within-run n = 21, total n = 63). The following results
level of 4.2% HbA1c (IFCC).
were obtained (data based on DCCT/NGSP values):
Icterus: No significant interference up to an I index of 60 (approximate
Within-run Mean SD CV
conjugated and unconjugated bilirubin concentration: 1000 µmol/L (60 mg/dL)).
% HbA1c % HbA1c %
Lipemia (Intralipid): No significant interference up to a triglyceride
Control Level 1 5.9 0.09 1.5
concentration of 500 mg/dL.
Control Level 2 10.5 0.11 1.1
Glycemia: No significant interference up to a glucose level of 55.5 mmol/L Human sample 1 5.2 0.04 0.8
(1000 mg/dL). A fasting sample is not required.
Human sample 2 9.0 0.11 1.2
Rheumatoid factors: No significant interference up to a rheumatoid
factor level of 750 IU/mL. Total Mean SD CV
Drugs: No interference was found using common drug panels.26 % HbA1c % HbA1c %
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated Control Level 1 6.0 0.12 1.9
hemoglobin, glycated albumin and labile HbA1c were found for the Control Level 2 10.5 0.21 2.0
anti-HbA1c antibodies used in this kit.
Human sample 3 5.6 0.07 1.3
For diagnostic purposes, the results should always be assessed in conjunction
with the patient’s medical history, clinical examination and other findings. Human sample 4 11.8 0.20 1.7

Measuring range for Whole Blood and Hemolysate Application Method comparison
Hemoglobin: 4-35 g/dL HbA1c (%) values for human blood samples obtained on a Roche/Hitachi
HbA1c: 0.3-2.5 g/dL cobas c 501 analyzer (y) were compared with those determined using
The technical limit in the instrument setting is defined as 0.9-3.1 g/dL the same reagent on a COBAS INTEGRA 800 analyzer (x) and on
due to the instrument factor for HbA1c (b = - 0.6; see above a Roche/Hitachi MODULAR P analyzer (x).
chapter cobas c 501 test definition). x = COBAS INTEGRA 800 analyzer, y = cobas c 501 analyzer
This corresponds to a measuring range of 2.3-18.9% HbA1c at a typical Sample size (n) = 109
hemoglobin concentration of 13.2 g/dL (IFCC values; corresponding Passing/Bablok28 Linear regression
values for DCCT/NGSP: 4.3-18.8% HbA1c). y = 0.9901x + 0.1097 y = 1.0033x + 0.0231
Lower detection limit τ = 0.9578 r = 0.9962
Hemoglobin: 0.5 g/dL The sample concentrations were between 5.0 and 12.6% (5.0 and 13.1%,
HbA1c: 0.1 g/dL DCCT/NGSP values).
A typical lower limit for the test range for %HbA1c may be calculated,
based on a given Hb concentration. Assuming a typical Hb concentration x = Roche/Hitachi MODULAR P analyzer, y = cobas c 501 analyzer
of 13.2 g/dL, the lower limit of the test range for %HbA1c is 0.8% (IFCC Sample size (n) = 93
values; corresponding value acc. to DCCT/NGSP is 2.9%). Passing/Bablok28 Linear regression
The lower detection limit represents the lowest measurable analyte y = 0.9837x + 0.1355 y = 0.9785x + 0.1967
level that can be distinguished from zero. It is calculated as the value τ = 0.9396 r = 0.9951
lying three standard deviations above that of the lowest standard
The sample concentrations were between 5.1 and 13.1% (5.0 and 12.9%,
(standard 1 + 3 SD, within-run precision, n = 21).
DCCT/NGSP values).
Expected values for Whole Blood and Hemolysate Application
Protocol 1 (acc. to IFCC): 2.9-4.2% HbA1c27 In addition, a comparison to a commercially available HPLC method was
performed. The HPLC method was standardized in conformance with
Protocol 2 (acc. to DCCT/NGSP): 4.8-5.9% HbA1c27
DCCT (Diabetes Control and Complications Trial).3,4
HbA1c levels above the established reference range are an indication of HPLC method
hyperglycemia during the preceding 2 to 3 months or longer. HbA1c levels Sample size (n) = 40
may reach 20% or higher in poorly controlled diabetes. Therapeutic action is
suggested at levels above 8%. Diabetes patients with HbA1c levels below 7% Passing/Bablok28 Linear regression
meet the goal of the American Diabetes Association.18 HbA1c levels below the y = 0.9349x + 0.4499 y = 0.9236x + 0.5666
established reference range may indicate recent episodes of hypoglycemia, τ = 0.9505 r = 0.9929
the presence of Hb variants, or shortened lifetime of erythrocytes.
The sample concentrations were between 5.3 and 11.9 % (5.4 and 11.8 %,
Each laboratory should investigate the transferability of the expected values to
DCCT/NGSP values).
its own patient population and if necessary determine its own reference ranges.

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04528123190V3

A1C-2
Tina-quant Hemoglobin A1c Gen.2
Specific performance data for Hemolysate Application References
Representative performance data on the analyzers are given below. 1. Goldstein DE, Little RR, Lorenz RA, Malone JI, Nathan D, Peterson CM.
Results obtained in individual laboratories may differ. Tests of glycemia in diabetes. Diabetes Care 1995;18:896-909.
Precision 2. Goldstein DE, Little RR. More than you ever wanted to know (but need to
Reproducibility was determined using human samples and controls in an know) about glycohemoglobin testing. Diabetes Care 1994;17:938-939.
internal protocol (within-run n = 21, total n = 63). The following results 3. The Diabetes Control and Complications Trial Research Group.
were obtained (data based on DCCT/NGSP values): The effect of intensive treatment of diabetes on the development
Within-run Mean SD CV and progression of long-term complications in insulin-dependent
% HbA1c % HbA1c % diabetes mellitus. N Engl J Med 1993;329:977-986.
5.9 0.13 2.2 4. Santiago JV. Lessons from the diabetes control and complications
Control Level 1
trial. Diabetes 1993;42:1549-1554.
Control Level 2 10.4 0.11 1.0
5. UK Prospective Diabetes Study (UKPDS) group. Intensive blood
Human sample 1 5.2 0.08 1.5 glucose control with sulfonylureas or insulin compared with conventional
Human sample 2 8.8 0.07 0.8 treatment and risk of complications in patients with type 2 diabetes
(UKPDS 33). Lancet 1998;352:837-853.
Total Mean SD CV 6. Flückiger R, Mortensen HB. Review: glycated haemoglobins.
% HbA1c % HbA1c % J Chromatogr 1988;429:279-292.
Control Level 1 6.0 0.12 2.0 7. Goldstein DE, Little RR, Wiedmeyer HM, England JD, McKenzie
Control Level 2 10.1 0.17 1.6 EM. Glycated hemoglobin: methodologies and clinical applications.
Human sample 3 5.6 0.09 1.6 Clin Chem 1986;32:B64-B70.
Human sample 4 11.7 0.18 1.5 8. Rohlfing CL, Wiedmeyer HM, Little RR, England JD, Tennill A,
Goldstein DE. Defining the relationship between plasma glucose
Method comparison and HbA1c. Diabetes Care 2002,25:275-278.
HbA1c (%) values for human blood samples obtained on a Roche/Hitachi 9. Bunn HF, Gabbay KH, Gallop PM. The glycosylation of hemoglobin:
cobas c 501 analyzer (y) were compared with those determined using relevance to diabetes mellitus. Science 1978;200:21-27.
the same reagent on a COBAS INTEGRA 800 analyzer (x) and on 10. Zander R, Lang W, Wolf HU. Alkaline haematin D-575, a new tool for the
a Roche/Hitachi MODULAR P analyzer (x). determination of haemoglobin as an alternative to the cyanhaemiglobin
x = COBAS INTEGRA 800 analyzer, y = cobas c 501 analyzer method. I. Description of the method. Clin Chim Acta 1984;136:83-93.
Sample size (n) = 109 11. Wolf HU, Lang W, Zander R. Alkaline haematin D-575, a new
Passing/Bablok28 Linear regression tool for the determination of haemoglobin as an alternative to the
y = 1.0322 x - 0.1447 y = 1.0268 x - 0.1082 cyanhaemiglobin method. II. Standardization of the method using
τ = 0.9619 r = 0.9979 pure chlorohaemin. Clin Chim Acta 1984;136:95-104.
12. Little RR, Wiedmeyer HM, England JD, Wilke AL, Rohlfing CL,
The sample concentrations were between 5.2 and 13.1 % (5.2 and 13.3 %,
Wians FH, et al. Interlaboratory standardization of measurements
DCCT/NGSP values).
of glycohemoglobins. Clin Chem 1992;38:2472-2478.
x = Roche/Hitachi MODULAR P analyzer, y = cobas c 501 analyzer 13. Frank EL, Moulton L, Little RR, Wiedmeyer HM, Rohlfing C, Roberts
Sample size (n) = 94 WL. Effects of hemoglobin C and S traits on seven glycated hemoglobin
methods. Clin Chem 2000;46 (6):864-867.
Passing/Bablok28 Linear regression 14. Chang J, Hoke C, Ettinger B, Penerian G. Evaluation and interference
y = 1.0000 x + 0.0740 y = 0.9996 x + 0.1047 study of hemoglobin A1c measured by turbidimetric inhibition
τ = 0.9504 r = 0.9968 immunoassay. Am J Clin Pathol 1998;109:274-278.
The sample concentrations were between 5.1 and 13.1 % (5.2 and 13.3 %, 15. Data on file at Roche Diagnostics.
DCCT/NGSP values). 16. Kobold U, Jeppsson JO, Duelffer T, Finke A, Hoelzel W, Miedema
K. Candidate reference methods for hemoglobin A1c based on
In addition, a comparison to a commercially available HPLC method was peptide mapping. Clin Chem 1997;43:1944-1951.
performed. The HPLC method was standardized in conformance with 17. Jeppsson JO, Kobold U, Finke A, Hoelzel W, Hoshino T, Miedema K,
DCCT (Diabetes Control and Complications Trial).3,4 Mosca A, Mauri P, Paroni R, Thienpont L, Umemoto M, Weykamp C.
HPLC method Approved IFCC reference method for the measurement of HbA1c in
Sample size (n) = 40 human blood. Clin Chem Lab Med 2002;40:78-89.
Passing/Bablok28 Linear regression 18. Martina WV, Martijn EG, van der Molen M, Schermer JG, Muskiet
y = 0.9490x + 0.4066 y = 0.9641x + 0.3276 FAJ. β-N-terminal glycohemoglobins in subjects with common
τ = 0.9544 r = 0.9936 hemoglobinopathies: relation with fructosamine and mean erythrocyte
age. Clin Chem 1993;39:2259-2265.
The sample concentrations were between 5.3 and 11.9 % (5.4 and 12.1 %, 19. Weykamp CW, Penders TJ, Muskiet FAJ, van der Slik W. Influence
DCCT/NGSP values). of hemoglobin variants and derivatives on glycohemoglobin
determinations, as investigated by 102 laboratories using 16
Analytical specificity for Whole Blood and Hemolysate Application methods. Clin Chem 1993;39:1717-1723.
Hb derivatives Labile HbA1c (pre-HbA1c), acetylated Hb, and 20. American Diabetes Association. Standards of Medical Care for patients
carbamylated Hb do not affect the assay results. with diabetes mellitus. Diabetes Care [Suppl] 18/1, 1995:8-15.
Hb variants Specimens containing high amounts of HbF (>10%) may 21. Sacks BW, Bruns DE, Goldstein DE, Maclaren NK, McDonald JM, Parrott M.
yield lower than expected HbA1c results. Guidelines and recommendations for laboratory analysis in the diagnosis
and management of diabetes mellitus. Clin Chem 2002;48:436-472.
Please note 22. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences
The IFCC and NGSP directed the manufacturers not to change their current in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-474.
HbA1c report out values until further decisions by the ADA/EASD/IDF 23. Miedema K. Influence of hemoglobin variants on the determination
working group on the HbA1c assay.29 This means that most countries should of glycated hemoglobin. Klin Lab 1993;39:1029-1032.
continue to report the established DCCT/NGSP values. If you are uncertain 24. Niederau CM et al. Interference of non glucose-adducts on the
of the situation in your country, please contact your local authorities to determination of glycated hemoglobins. Klin Lab 1993;39:1015-1023.
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