11.87M

2. Clinical Knowledge&Messurement Principle_V2.0

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2. Clinical Knowledge
& Messurement Principle
H-120 MCSR Training

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Definition
Name
Description
Hb
Hemoglobin
HbA
Main ingredients of Adult
Hemoglobin
HbA2
Minor ingredients of Adult
Hemoglobin
HbF
Fetal Hemoglobin
HbA0
Non-Glycosylated Hemoglobin
HbA1
Glycosylated Hemoglobin
HbA1a
Combination of HbA1 and Fructose
HbA1b Combination of HbA1 and G-6-P
HbA1c
Combination of HbA1 and Glucose

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HbA1c formation
Glucose enters red blood cells and reacts with
hemoglobin.

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HbA1c formation
Features:
1.
Parallel to blood sugar
values.
2.
The generation is slow.
3.
HbA1c cannot be easily
decomposed once they
occur.
4.
Less affected by HGB level
The life span of RBC is about 120 days. HbA1c can reflect the average blood glucose level
120 days before the measurement.
The time required for A1c to reach the midpoint of forming a new stable state level from the
initial level is 30-35 days. The recent changes in blood sugar have a greater impact on A1c
In the recent 30days, 31 -60 days, 61 -90 days, and 91 -120 days, the average blood
glucose level has an influence on the HbA1c of 50%, 25%, 15%, and 10%, respectively.

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Clinical significance of HbA1c
Preferred diagnostic marker for diabetes recommended by the
World Health Organization and many national diabetes societies
As a glycemic control indicator, it is the most effective and reliable
indicator for recent disease control of diabetes. It is also an
effective indicator for evaluating the management plan.
Used to predict the risk of complications in diabetes patients

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H 120/H120P measurement principle

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HbA1c test method
Test Method
HbA1c test
HPLC
(Gold standard)
Separation
Chemical
reaction
reaction
Affinity
chromatogr
change
HPLC ion
exchange
aphy
Disadvantages
1. An abnormal Hb
graph is displayed.
method
Charge
Benefits
2. HPLC is the method
with the highest
precision and accuracy.
3. The USA and Japan
use it as the reference
method for HbA1c.
Capillary
electrophoresis
Enzyme
Immunoturbidimet
method
ry
Can separate abnormal
Hb
Slow, expensive and complicated to use.
Borate affinity
chromatography
Not
affected
variants
Not only the N-terminal valine of β chain, but also the
glycation of other ends of β chain and the glycation of α
chain end are used to measure the glycated Hb. The
glycated hemoglobin is calculated through the empirical
algorithm and may be inaccurate. Cannot differentiate
between variants
Immunoturbidi
metry
It is not affected by
HbE, HbD, or HbCarb.
May be interfered by some mutations of HbS, C, and HbF.
Lipemia and icterus. Poor precision;
The variant cannot be separated; It is difficult to prepare
antibodies.
Enzyme method
Low cost and fast
response
Poor precision; Hemoglobin variants cannot be prompted.
Subject to lipemia and icterus. Subject to interference from
samples such as HbF and HbH.
by
Capillary
electrophor
esis
High cost, Susceptible to variant hemoglobin HbS, HbC,
HbD, HbE (different manufacturers are subject to different
degrees of interference) and other variants; H120 Variant
interference has been resolved

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HPLC
• HPLC: Different components carry different charges for separation
Hemolysis
processing
Hb
Hb
G Hb
G
Hb
Hb
Hb
Hb G Hb
Hb
Hb
The non-glycosylated Hb is positively charged and
bound to the functional group of the
chromatography column, and finally eluted.
Hb
Glycosylated Hb is eluted
first because glycosylation
has little positive charge.
HbA1 is separated based on
the difference between the
number of glycosyl groups
and the affinity of functional
groups of the
chromatography column.
s-A1c
A0
Variant
F
Hb
G
L-A1c
A1b
A1a
Hb
G
Detector

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Principle of High Performance Liquid Chromatography (HPLC)
Loading
Loa
ding
Sample preparation
Sample
preparation
Ultra-long
pressure
storage
technology
Sep
Separation
arati
on
Sample
Double
wavelengt
h
415nm/500nm
Reagent A
Pump 1
Detect
or
Injection
valve
Reag
ent B
Pump 2
reag
ent
B
A
Time
Figure 1 - High Pressure Gradient Elution
The dynamic proportion of reagent A and B. The elution
accuracy is 0.1%, which is twenty times higher than the
industry standard.
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Electronics Co., Ltd. All rights Reserved.
column
Consta
nt
temper
ature
room

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Summary
Understand the basic concept, formation, and clinical significance of HbA1c
Understand the different detection methods for HbA1c and their respective advantages and
disadvantages
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Understand the measurement principle of H-120 HPLC
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rights Reserved.

13.

Thank you!

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