Boditech School

- Distributor Training Program -

HbA1c
2013

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Diabetes
Definition
• Diabetes is usually a lifelong (chronic) disease in which there
are high levels of sugar in the blood, either because insulin
production is insufficient, or because the body's cells do not
respond properly to insulin, or both.

POCT
• Glucose monitoring
• HbA1c
• Microalbuminurea

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HbA1c
What is HbA1c?
Glycated hemoglobin, or glycohemoglobin.

How does it formed?

1. When sugar builds up in your blood during a diabetic

condition and it combines with hemoglobin (Hb).
2. Hb remains glycated and becomes HbA1c.
3. It stays that way until red blood cells die, which takes
about 12 weeks.

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HbA1c is highly glycated during diabetes

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Hb - Hemoglobin
HbA0: non-glycated hemoglobin.
HbA1: glycated hemoglobin
HbA1a1: Glycation with Fructose 1, 6 diphosphate
HbA1a2: Glycation with Glucose 6 phosphate
HbA1b: Glycation with an unknown reaction partner

HbA1c - Glycation with D-glucose

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Why is HbA1c test important?
The amount of HbA1c reflects the blood sugar
levels for the last several weeks, typically
encompassing periods of 3-4 months.

When do Drs. order HbA1c test?

• as a screening tool for diabetes

• as a routine test if you have diabetes
4-5.6% 5.7-6.4% Above 6.5%
normal risk of diabetes diabetes

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 What is the meaning of % for HbA1c

Amount of Glycated Hemoglobin

% of HbA1c =
Amount of Total Hemoglobin

Need to measure both amounts in human blood once

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i-Chroma HbA1c

i-Chamber Cartrige

사용 전 (최초 pouch 개봉시)

사용 후 (Cover opened)

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i-Chroma HbA1c
For measuring glycated hemoglobin in human blood specimen

 Competition Immunoassay on the test device

HbA1c(in sample)
Sample
fluorescence
fluorescence
Competition HbA1c
Peptide

Test line Control line

Flow direction
HbA1c antibody

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i-Chroma HbA1c
For measuring Total hemoglobin in human blood specimen

 Hemoglobin assay
Sample Met-hemoglobin
potassium ferricyanide

525nm light

Measurement

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i-Chroma HbA1c
Specifications Data
Product name iChroma™ HbA1c
Intended use Quantitative test of HbA1c % in human blood
CV % <10%
Correlation R2 >= 0. 9
Reaction time 12min
Detection Range,Ref. value 4 – 15% , 6.5%
Sample type Whole blood
Sample volume 5 uL
DB volume -
Stability 20 months, 2 ~ 8℃
Test device : 4 ~ 30℃
Storage condition
Detection buffer : 2 ~ 8℃

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Test procedure
Preparation - 1

Check the materials before HbA1c Testing

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Test procedure
Preparation - 2
[Cartridge]
1. Insert a cartridge into a i-Chamber slot for
stabilizing the reaction temperature.

* Check if the temperature of i-Chamber is

30˚C with a centigrade thermometer

2. Keep the cartridge in the slot for 5~30

minutes.

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Test procedure
Preparation - 3
[Reaction Buffer]
1. Take 100uL of detection buffer from plastic
amber vial.

2. Add the buffer into a pre-dispensed

Hemolysis buffer tube.

* Pipetting should be handled carefully to

avoid the error from a burble or lower
volume.

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Test procedure
Blood collection
1. Collect 5 uL of whole blood from
fingertip or tube blood with capillary.

*Be sure to check how to use capillary.

* Finger tip blood: Prick the finger

with a lancet and discard the first
drop. Collect 5uL from the second
drop with a capillary.

* Tube blood: Whole blood collected

in a tube containing heparin or EDTA
can be used. Mix the whole blood by
inverting the tube.

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Test procedure
Blood collection
2. Put the capillary contains blood sample
into the tube prepared in “reaction
buffer preparation step”.

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Test procedure
Sample processing
1. Shake the tube 15 times strongly in order
that the blood come out from capillary.

2. Pull the cartridge halfway out until HbA1c

well is seen fully from i-Chamber slot after
5 minutes since cartridge incubate in i-
Chamber.
* Do NOT take the cartridge out completely.

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Test procedure
Sample processing
3. Apply 150uL(75 ul x 2 times) of the mixture
into the Hb well using a pipette

* When bubble appears in Hb window, try the

test again because mislead value may come.

* To avoid bubble, please lean the pipette

about 45˚-60˚ on the side wall.

* Please wipe out the dirts on the surface

of Hb window, if any.

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Test procedure
Sample processing
Apply 75ul of the mixture into the A1c well and
re-insert the cartridge in i-chamber

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Test procedure
Reading
1. Keep the cartridge in i-Chamber for 12
minutes before scanning by i-CHROMA
Reader

* Watch out so as not to touch to the

surface of window 1 when the cartridge
comes out.
* Take care not to cause shock and leakage
of loading mixture on the well.

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Test procedure
Reading
2. The cover of window 2 is automatically
opened when the cartridge is inserted into
a holder of i-CHROMA reader.

* If the cartridge cannot be inserted to the

holder, you can solve the problem through
press a cover of window 2 with hand.

3. Press “Select” button, and then read the

result on the LCD screen.

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 Accuracy of i-Chroma HbA1c
• The accuracy of HbA1c(New) with i-Chamber is remarkably improved according to keeping
temperature through incubation the cartridge in the chamber.

• Test Value

• Recovery

7% Patient Sample

Cartridge
Products Data Run1 Run2 Run3 Run4 Run5 Run6 AVG CV(%)
Temp

Value 7.3 6.7 6.9 6.8 6.8 6.9 6.9 3.2

20~25℃
HbA1c Recovery(%) 104.9 96.1 98.0 97.3 97.1 98.9 98.7
(New)
Value 6.8 6.7 7.3 7.4 6.7 7.0 7.0 4.6
37℃
Recovery(%) 96.7 95.4 104.0 105.9 95.3 100.0 99.5

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Accuracy of i-Chroma HbA1c
Reproducibility(Between run, Interassay CV%)
of i-CHROMA HbA1c(new) with i-Chamber
G7 Tester
No. AVG CV(%)
(Reference) Tester 1 Tester 2 Tester 3
1 5 5.19 5.5 4.99 5.23 4.96
2 5.1 5.35 5.12 5.29 6.18 2.71
3 5.5 5.14 5.4 5.77 7.03 3.76
4 5.6 5.56 5.5 5.95 5.44 5.85
5 5.9 6.27 6.28 5.99 7.29 0.94
6 6 6.22 6.52 6.41 6.69 1.19
7 6.3 6.4 6.18 5.97 7.13 2.44
8 6.3 6.59 6.36 6.26 5.25 2.28
9 6.4 6.8 6.39 6.66 5.67 4.31
10 6.7 6.36 6.69 6.35 8.96 4.84
11 6.8 6.76 6.71 6.6 6.62 3.15
12 6.9 7.17 7 6.72 7.65 3.29
13 7 7.09 6.98 7.32 6.47 3.01
14 7.1 6.73 7.23 7.13 6.18 3.52
15 7.3 7.33 7.32 7.21 7.78 2.10
16 7.3 7.43 7.02 7.2 6.38 2.38
17 7.8 7.77 7.36 7.82 6.96 3.24
18 8 7.64 7.96 7.74 6.40 2.62
19 8.5 8.9 8.71 8.94 8.85 1.40
20 9 8.56 8.91 9.42 7.22 2.85

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Accuracy of i-Chroma HbA1c
• Difference of Correlation(R²) among testers
• Tester 1 ; R² = 0.9333
• Tester 2 ; R² = 0.9539
• Tester 3 ; R² = 0.9462
10 y = 0.9203x + 0.5741
R² = 0.9333
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(Tester 1)
8 y = 0.9197x + 0.5721

7 R² = 0.9539
(Tester 2)
6 Tester 1

y = 1.0029x + 0.0425 Tester 2

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R² = 0.9462 Tester 3
HbA1c New (%)

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(Tester 3) 선형 (Tester 1)
3 선형 (Tester 2)
선형 (Tester 3)
2

0
0 2 4 6 8 10
Tosoh G7 HBA1c (%) HPLC

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Comparison study of i-Chroma HbA1c
• High consistency is shown ; Comparative result , R² = 0.98 in comparison
with Tosho G7(HPLC)

i-CHROMA HbA1c vs Tosho G7

16.0

14.0 y = 0.9939x + 0.0038

12.0 R² = 0.9835
i-CHROMA HbA1c (%)

10.0 N= 150
8.0

6.0

4.0

2.0

0.0
0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0

Tosoh G7 HbA1c (%)

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Comparison study of i-Chroma HbA1c
• Correlation(R2) between i-CHROMA HbA1c and other POCT products is 0.933(Nycocard)
and 0.932(Labonacheck), respectively
Tosoh
No HbA1C “N” “L” "N" with i-CHAMBER HbA1c
G7
1 5.5 5.6 5.5 4.6 14
2 5.5 5.6 5.6 4.9 12 y = 0.9728x + 0.235

i-CHROMA HbA1c (%)

3 5.8 5.7 5.6 5.3
10
4 6.0 5.9 6.3 5.5 R² = 0.9335
5 6.0 6.3 5.8 5.6 8
6 6.1 5.7 6.1 5.4 6
7 6.3 6.7 6.1 5.6
8 6.4 6.5 6.2 5.4 4
9 6.7 6.3 6.5 6.4 2
10 6.7 6.4 6.3 5.6
0
11 6.9 6.9 7.0 6.0
0 2 4 6 8 10 12 14
12 6.9 6.5 6.4 5.8 “N” HbA1c (%)
13 7.1 6.2 6.8 6.2
14 7.2 6.5 6.6 6.3
15 7.3 6.5 6.6 6.1 "L" with i-CHROMA HbA1c
16 7.5 7.1 6.9 6.2
17 8.0 7.7 6.6 6.7 14
18 8.1 7.3 7.7 7.1 12 y = 0.8603x + 1.4467
19 8.2 7.4 7.2 7.2
i-CHROMA HbA1c(%)

20 8.6 7.5 7.9 7.0 10 R² = 0.9325

21 8.7 7.8 7.7 7.8 8
22 8.9 8.4 8.0 7.5
6
23 9.0 8.2 8.8 7.7
24 9.4 8.3 8.7 7.5 4
25 9.4 8.9 8.5 8.3 2
26 11.6 10.3 9.8 11.5
0
27 12.4 11.0 10.6 10.7 0 2 4 6 8 10 12 14
28 13.2 11.6 11.9 11.0 “L” HbA1c(%)

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