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EU rev. G8S-0500705
Safety Precautions
To help protect you and/or your property from potential damage and ensure personal
safety, please read this IFU thoroughly before using the product.

Automated Glycohemoglobin Analyzer HLC-723®G8 [NOTATIONAL CONVENTION]

Notation Explanation
Indicates a hazard with a low level of risk which, if not avoided,
CAUTION
could result in minor or moderate injury.

Instructions For Use CAUTION

■ Use only in well ventilated areas
G8 HSi Elution Buffer No. 1 (S) In case of insufficient ventilation, spilled solvents can cause harm.
■ First Aid
No. 2 (S) Skin exposure
Wash exposed area with plenty of soap and water.
No. 3 (S) Eye exposure
Open eyes as wide as possible and wash with clean water for at least 15 minutes.
Immediately call for medical attention.
Ingestion
Please wash mouth with excess water and immediately call for medical attention.
■ Do not spill solvents
Spillage and leakage can cause fire, electric shock, poisoning, injury, and corrosion.
Wear appropriate protective gear when cleaning up a spill.
■ Wear eye protection and protective gloves
Organic acids are harmful and should not come in direct contact with the skin.
■ Handle package with care
Inappropriate handling may cause rupturing and/or spattering of the product.
■ Only use this product as intended
This product is intended for in vitro diagnostic use for the measurement of HbA1c in blood
specimens.
■ Proper disposal
Dispose in accordance with local laws and regulations.

NOTE
Keep this IFU with the product for future reference.

Attention: This IFU complies with IVD directive 98/79/EC and is intended for use by
customers operating in a member state of the European Union.
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Revised in 11/2019
CONTENTS
1) Introduction
1) Introduction .................................................................................................................................. 4 The G8 Elution Buffer HSi is designed exclusively for use with the Tosoh Automated
2) Before Use ................................................................................................................................... 4 Glycohemoglobin Analyzer HLC-723G8 (referred to as HLC-723G8 in the IFU), which is based
3) Warnings and Precautions .......................................................................................................... 4 on the principle of High-Performance Liquid Chromatography assay. It is not designed for and
4) Content ........................................................................................................................................ 4 should never be used with any other type of system.
5) Related Components ................................................................................................................... 5
6) Storage and Stability ................................................................................................................... 5 The G8 Elution Buffer HSi is intended for in vitro diagnostic use for the measurement of
7) Specimens ................................................................................................................................... 5 haemoglobin A1c (HbA1c) in blood specimens.
8) Assay Principle ............................................................................................................................ 5
9) Installation .................................................................................................................................... 5 Glycohaemoglobin (GHb) is a general term for complexes where the whole blood glucose is
10) Assay Procedures........................................................................................................................ 6 non-enzymatically bound to the α or β-chains of human haemoglobin. Within these complexes,
11) Precautions for Use ..................................................................................................................... 6 HbA1c, which is a complex of glucose and the N-terminus of the β-chain, is the most
12) Reference Data (from the HLC-723G8 Assay Examples) .......................................................... 7 quantitatively prevalent.
13) Measurement Values................................................................................................................... 7 HbA1c is non-enzymatically synthesized in two steps.
14) Evaluation of Results ................................................................................................................... 7 Step 1: The glucose aldehyde group reacts with the free amino group on the valine
15) Expected Values .......................................................................................................................... 7 comprising the N-terminus of the β-chain to form the Schiff base (labile HbA1c
16) Performance Characteristics ....................................................................................................... 8 (L-A1c)).
17) Precision ...................................................................................................................................... 9 Step 2: Stable ketoamine (stable HbA1c (s-A1c)) is then formed by a reaction known as the
18) Interferences ................................................................................................................................ 9 amadori rearrangement.
19) Specimen Collection and Handling ........................................................................................... 11 Due to the fact that L-A1c (the intermediate product of this reaction) changes rapidly in response
20) References ................................................................................................................................ 11 to the changes in whole blood glucose concentrations, s-A1c is now generally used to measure
HbA1c. It provides a better indication of the average glucose level over the past couple of
months because it does not fluctuate in response to physiological factors.

In the past, it was difficult to achieve quick separation of L-A1c and s-A1c on a column; therefore
sample pre-treatment to remove L-A1c was necessary before assaying s-A1c. The new Tosoh
Automated Glycohemoglobin Analyzer HLC-723G8 features quick separation of L-A1c and
s-A1c on the column (1.0 min.), with no need for sample treatment when assaying s-A1c.
2) Before Use
Inspect the packaging and the exterior of the aluminium pack for any signs of damage before
use. If any damages are visible, contact your local Tosoh sales representative.

Confirm the following document is included in the package.

• Instructions For Use 1 copy (in each box)
3) Warnings and Precautions
1) This product is for in vitro diagnostic use only.
2) This product is intended for use on the Tosoh Automated Glycohemoglobin Analyzer
HLC-723G8 Standard Analysis Mode only.
3) Do not use this product beyond the expiry date.
4) Content
The following types of elution buffers are designed for exclusive use with the HLC-723G8.
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Catalogue No. Description Contents (per one 4) Gently squeeze the elution buffer pack by hand to remove all excess air, and tighten
Pack) cap firmly into place to create a vacuum that will prevent air from entering during
0021800 G8 Elution Buffer HSi No. 1 (S) 800 mL operation.
0021801 G8 Elution Buffer HSi No. 2 (S) 800 mL
5) Press the MAINTE key and then the REAGENT CHANGE key.
0021802 G8 Elution Buffer HSi No. 3 (S) 800 mL
6) Press the keys of the reagents to be replaced on the REAGENT CHANGE screen.
G8 Elution Buffer HSi is an organic acid buffer. Each contains less than 0.05 % sodium azide as a 7) Press the CHANGE key. A confirmation message will be displayed.
preservative. If everything is all right, press the OK key.
10) Assay Procedures
Assay mode : Standard Analysis Mode
Refer to the HLC-723G8 Operators’ manual for detailed instructions.
Applicable analysers : Tosoh Automated Glycohemoglobin Analyzer HLC-723G8
5) Related Components 11) Precautions for Use
Catalogue No. 1) Carefully read the instructions contained in this IFU and related Instructions For Use
Hemoglobin A1c Calibrator Set 0018767 provided with HLC-723G8, TSKgel G8 HSi, Hemoglobin A1c Calibrator Set, G8 HbA1c
G8 HbA1c Calibrator Set (S) 0023528 Calibrator Set (S), Hemoglobin A1c Control Set, HbA1c Diluting Solution and HSi
Hemoglobin A1c Control Set 0021974 Hemolysis & Wash Solution.
TSKgel® G8 HSi 0021498
2) The G8 Elution Buffer HSi is designed exclusively for use in combination with the
HSi Hemolysis & Wash Solution (L) 018431L
HSi Hemolysis & Wash Solution (LL) 0019550 HLC-723G8 analyzer system, TSKgel G8 HSi and HSi Hemolysis & Wash Solution
HbA1c Diluting Solution 0023503 indicated below, never in any other combination.
• Tosoh Automated Glycohemoglobin Analyzer HLC-723G8
6) Storage and Stability • TSKgel G8 HSi
• HSi Hemolysis & Wash Solution (L), (LL)
All unopened materials will remain stable until the expiry date on the label when stored at 4 to
3) Always use the G8 Elution Buffer HSi in combination with the TSKgel G8 HSi of the identical
30 °C.
lot number. The column lot number is indicated by a single uppercase alphabetical
The expiry date indicated on the package box and aluminium pack labels.
character (A, B, etc.) on the label of column box. The elution buffer label displays an
When stored at 4 to 30 °C, each buffer is stable for 3 months after opening.
alphabetic character corresponding to the column lot number, as shown below. Note that
7) Specimens the G8 Elution Buffer HSi No. 3 is compatible with all lots.
Whole blood samples.
8) Assay Principle
The HLC-723G8 is based on the High-Performance Liquid Chromatography (HPLC) principle.
Cation exchange columns employ the differences in ionic interactions between haemoglobin
components to separate them into a total of six fractions in a span of 1.0 min.
A step gradient elution is used to separate and assay HbF, s-A1c, and Total A1, with three types 4) Return the elution buffer to room temperature before use.
of G8 Elution Buffer HSi (G8 Elution Buffer HSi No.1, 2, and 3 (S)) of different salt 5) When first opening the aluminium pack to insert the suction tube, gently squeeze the elution
concentrations. buffer pack by hand to remove all excess air, and tighten bottle cap firmly to create a
9) Installation vacuum that will prevent air from entering during operation.
6) Never use reagents that have exceeded the expiry date printed on their labels. Assay
Be sure to read the IFU included with the G8 Elution Buffer HSi, TSKgel G8 HSi, Hemoglobin
results for expired reagents will not be reliable. Also note that reagents must be used within
A1c Calibrator Set, G8 HbA1c Calibrator Set (S), Hemoglobin A1c Control Set, HbA1c Diluting
three months after opening (provided that the container is correctly maintained in vacuum
Solution and HSi Hemolysis & Wash Solution as well as the HLC-723G8 Operators’ manual.
state).
7) When there is leftover reagent in the aluminium pack that has to be removed from the
1) Press the Stop key on the HLC-723G8 system operation panel to enable the
analyser and stored, again gently squeeze the elution buffer pack by hand to remove all
STAND-BY state.
excess air, and tighten the bottle cap firmly to create a vacuum that will prevent air from
2) Remove the cap from the elution buffer pack.
entering during storage and store between 4 and 25 °C.
3) Make sure to insert the tube matching the colour of the elution buffer pack being
8) Always replace with an aluminium pack when elution buffer is almost empty. Avoid refilling
installed.
leftover elution buffer into aluminium pack as this can produce unreliable assay results.
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9) For safe waste disposal, it is recommended that each laboratory complies with established
laboratory procedures and local, state and federal regulations.
NGSP=0.09148 x FCC 2.152＋
12) Reference Data (from the HLC-723G8 Assay Examples)
The above categories in NGSP units can be converted to those in IFCC units below by
Format 100 Control Format 100 Sample applying the above equation.
HbA1c (s-A1c) >/= 48 mmol/mol threshold to diagnose diabetes
HbA1c (s-A1c) 39 - 47 mmol/mol categories of increased risk for diabetes
(prediabetes)
The above reference range (non-diabetic) is also converted as:
HbA1c (s-A1c) : 20 - 42 mmol/mol.
16) Performance Characteristics
1) Dilution Linearity
The tests were performed with Hemoglobin A1c Control diluted with various percentage
factors to determine the effect on the s-A1c (%) value. Linearity was observed for the Total
Area in the 500 to 2700 range, and measurement in the range of 700 to 2500 is
recommended to give more reliable results.

Total Area s-A1c (%)

324.50 9.4
652.54 9.9
939.78 9.9
1199.05 9.9
1552.53 10.0
1833.12 10.0
2104.36 10.1
2321.57 10.1
13) Measurement Values 2610.74 10.1
2858.85 10.2
Measurement values (%) indicate the percentage of each peak in relation to the Total Area
(excluding the front peak (FP)). Note the minimum unit of measurement displayed is 0.1 %.
2) Recovery
14) Evaluation of Results The table below shows the recovery ratios for two types of samples mixed in different
Quality Control percentage factors.
In order to monitor and evaluate the precision and analytical performance, it is recommended
that control samples should be run daily. If one or more control sample value(s) is out of the High s-A1c Low s-A1c Measurement Theoretical Recovery
sample (ratio) Sample (ratio) value s-A1c (%) value s-A1c (%) Ratio (%)
− −
acceptable range, it is necessary to investigate the validity of the calibration curve before 0 10 2.7
reporting patient results. Standard laboratory procedures should be followed in accordance to
the strictest regulatory agency under which the laboratory operates. 1 9 4.7 4.7 100.0
2 8 6.5 6.7 97.0
15) Expected Values 3 7 8.4 8.5 98.8
According to the American Diabetes Association (Reference 1): 4 6 10.2 10.3 99.0
HbA1c (s-A1c) >/= 6.5 % threshold to diagnose diabetes 5 5 11.9 12.2 97.5
HbA1c (s-A1c) 5.7 - 6.4 % categories of increased risk for diabetes 6 4 13.6 13.9 97.8
(prediabetes) 7 3 15.3 15.5 98.7
The reference range (non-diabetic) is reported as below in Reference 2. 8 2 17.0 17.3 98.3
HbA1c (s-A1c): 4.0 - 6.0 % (mean 5.0 %, SD 0.5 %) 9 1 18.8 19.0 98.9
10 0 20.1
− −
The values referred to within this IFU have been determined with a NGSP certified method. It
is known that the relationship between HbA1c results from the NGSP network (%) and the
IFCC network (mmol/mol) is expressed by using the following equation (Reference 3).
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3) Correlation 1) No effect was observed below 10 g/L for glucose, 0.25 g/L for sodium cyanate and
The correlation between the s-A1c (%) values for the HLC-723G8 and HLC-723G7 analysers 0.20 g/L for acetaldehyde.
is shown in the graph below. This method is certified by National Glycohemoglobin
Standardization Program (NGSP). Glucose

The correlation between the s-A1c (%) for HLC-723G8 and HLC-723G7 analyzers.

Sodium cyanate

17) Precision
1) Intra-assay (n=10)
The intra-assay precision (n=10) factors of the s-A1c (%) for three types of control samples
are shown below.

Low Medium High

Acetaldehyde
Mean 5.47 7.68 9.90
SD 0.05 0.06 0.00
CV % 0.91 0.78 0.00

2) Inter-assay (n=10)
The inter-assay precision (n=10) factors, measured once a day for a 10-day period, of the
s-A1c (%) for two types of control samples are shown below.

Low High
Mean 5.52 9.88
SD 0.04 0.06
CV % 0.72 0.61
18) Interferences
A substance is considered to interfere when the recovery of a known specimen falls outside a
10 % acceptability range. 2) No effect was observed for the following agents up to the concentrations indicated in the
Concentrations of interfering substances in this IFU are in units of g/L. table.

Free bilirubin to 0.19 g/L

Conjugated bilirubin to 0.20 g/L
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Chyle to 2,130 Formazine turbidity

Aspirin to 0.50 g/L

3) Haemoglobin variants (including HbS, HbE, HbC and HbD) and Thalassemias may give
false HbA1c results and the HbA1c values should not be reported. Typically, in the
presence of HbE, a small peak may be observed behind the s-A1c peak.
4) Low concentrations of haemoglobin may cause the TOTAL AREA of the chromatogram to
be < 500 and HbA1c values should not be reported.
5) High concentrations of haemoglobin may cause the TOTAL AREA of the chromatogram to
be > 2700 and HbA1c values should not be reported.
6) HbA1c may not correspond to average blood glucose in patients whose red blood cell
turnover is increased or decreased. Clinical interpretation has to be done with care.
7) Haemolysed samples do not interfere with HbA1c assay results.
19) Specimen Collection and Handling
A whole blood sample is required for the assay.
No special preparation is necessary. A venous blood sample is collected aseptically. The blood
samples in primary tubes, which contain EDTA, NaF, Heparin or Citric acid may be stored at TOSOH CORPORATION
25 °C for 24 hours before analysis or at 4 °C for 14 days. BIOSCIENCE DIVISION
Shiba-Koen First Bldg.
3-8-2, Shiba, Minato-ku, Tokyo 105-8623, Japan
20) References Phone: +81 3 5427 5181 Fax: +81 3 5427 5220
1) American Diabetes Association. Standards of Medical Care in Diabetes - 2013. Diabetes
Care 2013; 36 (Suppl. 1), S11-66. TOSOH EUROPE N.V.
Transportstraat 4
2) American Diabetes Association. Standards of Medical Care for Patients with Diabetes B-3980 Tessenderlo, Belgium
Mellitus. Diabetes Care 2000; 23 (Suppl. 1), S32-42. Phone: +32 13 66 88 30 Fax: +32 13 66 47 49
3) Geistanger A, Arends S, Berding C, Hoshino T, Jeppsson J-O, Little R, Siebelder C and
Weykamp C, Statistical Methods for Monitoring the Relationship between the IFCC
Reference Measurement Procedure for Hemoglobin A1c and the Designated Comparison This manual may not be reprinted or copied in whole or in part without written consent of Tosoh
Methods in the United States, Japan, and Sweden. Clinical Chemistry, 54:8, 1379-1385 Corporation. The contents of the manual are subject to change without notice.
(2008).
“HLC”, “HLC-723” and “TSKgel” are the registered trademarks of Tosoh Corporation in Japan, etc.

Symbols on the product labels

Actual manufacturing site For specified column lot only