HbA1C

Hemoglobin A1C – HbA1C immunturbidimetric Latex-Test

Cat.No Package Size Additional Reagents

Calibration Set: Cat. No. 270 410 (2 x 0.5 ml)
R1 (R1a+R1b) => Control Set: Cat. No. 270 510 (2 x 0.5 ml)
R2 = 10 mL
23,5 mL
HBA55S94
Lysing Reagent = 25 ml SAMPLES
Collect venous blood with EDTA.
GENERAL Storage and Stability
The glycemic control in diabetes mellitus is performed Hemoglobin A1c in whole blood with EDTA
5
mainly by the determination of Glucose, but also through is stable for one week at 2-8°C.
quantitative determination of Hemoglobin A1c (HbA1c) in
human blood : HbA1c is an indication for the actual glucose
levels over the preceding 3 months, being formed To determine HbA1c, a hemolysate must be prepared
continuously by adduction of glucose to the hemoglobin from each sample:
beta chain. It was shown that HbA1c in diabetic subjects 1. Dispense 250µl of Lysing Reagent into cups or tubes and
can be elevated 2-3 fold over normal and on the other label as Controls, Patients, etc.
hand approaches normal values when they are under 2. Add 20ul of well mixed (!) whole blood samples respectively
metabolic control. of Calibrators and Controls .
The procedure under consideration is based on the direct (Note: Calibrators and Controls have to be treated exactly like
determination of HbA1c through enzymatic/colorimetric the patient samples!)
reaction . 3. Let incubate at room temperature for minimum 10 min.
Stability:
Hemolysates may be stored up to 1 day at 2-8°C
PRINCIPLE
Hemolyzed blood is used as sample material. Through
protease attack glycated valines are released and are
PRECAUTIONS
subject to further enymatic reaction with fructosyl valine
1. The reagent is for in vitro diagnostic use only.
oxidase (FVO) . The result is a measureable quantitative
2. All human specimens should be regarded as potentially
amount of hydrogen peroxide, which is determined through
biohazardous. Therefore, universal precautions should be
colorimetric reaction with a chromogen compound.
used in specimen handling (gloves, lab garments, avoid
The reaction product is proportional to the amount of HbA1c aerosol production, etc.)
and is measured as absorbance A. The HbA1c value is
derived from a calibration curve.
ANALYTICAL PROCEDURE
REAGENTS This reagent is made esp. for use on automated
analyzers. Applications are available on request.
R1a: MES-Buffer (pH 7.0) 5.0 mmol/l Following (as an example) you find the instrument
Protease ≥ 3,5 kU/ml setting for the Hitachi 717 which is using the "2-reagent
Triton-X 100 0.5 %l procedure"
R1b: MES-Buffer (pH 6.3) 1.0 mmol/l
R2: TRIS-Buffer (pH 8.0) 5.0 mmol/l
Fructosyl Valine Oxidase ≥ 9,5 kU/l ASSAY CODE [2-POINT]:[24]-[40]
Peroxidase ≥ 8,5 kU/l SAMPLE VOLUME [20] [20]
Chromogen ≥ 0.7 mmol/l R1 VOLUME [128] [20] [NO]
R2 VOLUME [56] [20] [NO]
WAVELENGTH [ 800 ] [700]
Storage CALIBRATION [LINEAR] [0]-[0]
STD (1) CONC-POS [**] [*]
Store all reagents refrigerated at 2-8°C. Unopened r eagents STD (2) CONC-POS [**] [*]
are stable up to the expiration date printed on the labels. STD (3) CONC-POS [0] [0]
STD (4) CONC-POS [0] [0]
STD (5) CONC-POS [0] [0]
Preparation of Reagents STD (6) CONC-POS [0] [0]
UNIT %
3-reagent-procedure: SD LIMIT [999]
R1a, R1b and R2 are ready for use when the 3-reagent- DUPLICATE LIMIT [32000]
SENSITIVITY LIMIT [0]
procedure is applied. ABS LIMIT (INC/DEC) [32000]-[INCREASE]
Stability after opening : PROZONE LIMIT [0] [LOWER]
EXPECTED VALUE [*]-[*]
At least 3 months when contamination is avoided PANIC VALUE [-32000]-[32000]
INSTRUMENT FACTOR [1.00]
2-reagent-procedure: * determined by user
R1: Mix 7 parts R1a with 3 parts R1b ** Conc. of calibrators
Stability:
At least 2 weeks when contamination is avoided

Important Note: R1b and R2 are light sensitive !

L-S 12/2010 page 1/2
CALCULATION INTERFERENCES
HbA1c results are determined using calibration based on a 1. Bilirubin to 15mg/dL, ascorbic acid to 10mg/dL,
suitable mathematical procedure and the Inmesco triglycerides to 3000mg/dL, Glucose to 4000mg/dL,
Calibration Set. carbamylated Hb to 5mmol/L and acetylated Hb to
For the 2-reagent procedure the calibration is stable for at 5.0mmol/L do not interfere in the assay.
least 1 day. 2. It has been reported that results may be inconsistent in
For the 3-reagent procedure the calibration is stable for at patients who have the conditions like opiate addiction,
least 1 week. lead-poisoning, alcoholism, ingestion of large doses of
Check this by using Inmesco's Control Set. aspirin.

.
QUALITY CONTROL REFERENCES
We suggest the use Inmesco Hemoglobin A1c Control Set 1. Trivelli, L.A., Ranney, H.M., and Lai, H.T., New Eng. J.
with assayed values respectively ranges. Med. 284,353 (1971).
2. Gonen, B., and Rubenstein, A.H., Diabetologia 15, 1
(1978).
3. Gabbay, K.H., Hasty, K., Breslow, J.L., Ellison, R.C.,
EXPECTED VALUES
Bunn, H.F., and Gallop, P.M., J. Clin. Endocrinol.
Recommended Values are
Metab. 44, 859 (1977).
< 6% for non-diabetics
4. Bates, H.M., Lab. Mang., Vol 16 (Jan. 1978).
6 - 9% for diabetics under perfect glycemic control
5. Tietz, N.W., Textbook of Clinical Chemistry,
Up to 20% for diabetics out of glycemic control
Philadelphia, W.B. Saunders Company, p.794-795
Note: (1999).
Each laboratory should establish its own expected values. The 6. Corielo, A., et al, Diabetologia 22, p. 379 (1962).
given values can only be an average indication . 7. Goldstein, D.E., et al, Clin. Chem. 32, pp. 364-370
(1986).
8. Fluckiger, R., et al, Med Intelligence 304 pp. 823-827
LIMITATIONS (1981).
9. Nathan, D.M., et al, Clin. Chem. 29, pp. 466-469
1. Results may be inconsistent in patients e.g.with opiate
(1983).
addiction, lead-poisoning, alcoholism, ingestion of
10. American Diabetes Association: Clinical Practice
large doses of aspirin.
Recommendations, Diabetes Care 24 (Suppl. 1):
2. Elevated levels( > 10%) of HbF may lead to
pp. 33-S55, (2001).
underestimation of HA1c.
3. Hemoglobin variants HbS, HbC and HbE do not
interfere in this assay. There is also no interference by
labile intermediates , and uremia does not interfere, SYMBOLS USED
too.

For in vitro diagnostic medical use

PERFORMANCE CHARACTERISTICS
1. Linearity: The Hemoglobin A1c assay range is 2.0%
to 16.0%. Results in this range can be reported and Batch Code
used directly.
2. Correlation: A study using 40 human specimens
between this HA1c procedure and the reference
Use by
method yielded a correlation coefficient of 0.9874 and a
linear regression equation of y = 1.021 x + 0.014
3. Precision: Temperature limitation
Within Run: The intra assay precision was established
by assaying blood with two Hemoglobin A1c levels
twenty times each.

Level Mean % C.V.

Medium 5.7 1.0
High 10.3 0.7

L-S 12/2010 page 2/2