This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197219007** Revised: June 2018 (4th edition)* Revised: January 2017 (3rd edition)

This package insert must be read carefully prior to use.
Glycohemoglobin A1c assay kit(Classification No.: 30168000)
L typethe absorbance of this complex.
The percentage of total Hb existing as HbA1c is
calculated from the concentrations of HbA1c andHb.

NORUDIA N HbA1c

HbA1c:Protease
HbA1c ────▶ Glycosylated dipeptide

General Precautions
1. This product is for in vitro diagnostic use, and mustnot be used for any other purposes.
2. Clinicians should make a comprehensive clinicaldecision based on assay results in conjunction withclinical symptoms and other examination results.
3. This product should be used only as directed in thispackage insert. Reliability of results cannot be
guaranteed if there are any deviations from theinstructions in this package insert.
4. If the reagent accidentally comes in contact witheyes and/or mouth, rinse immediately with ample
water as first aid, and consult the doctor if required.
5. Carefully read the operating instructions for eachtype of automated analyzers prior to using thisproduct. Parameters for each type of analyzers areavailable, and can be requested from SEKISUIMEDICAL CO., LTD. if required.
6. Perform a quality control test prior to assay toensure accuracy.

FPOX
Glycosylated dipeptide + O2 + H2O ────▶
Glucosone + Dipeptide + H2O2
POD
H2O2 + Coloring agent ────▶ Coloration
2. Features
1) This product is a reagent that specificallymeasures HbA1c-derived glycosylated dipeptide
by the enzymatic method.
2) Liquid reagents, ready-to-use.
3) Reaction cells are hardly contaminated.
4) Analytical results are not affected by unstable
HbA1c or modified Hb (such as carbamylated oracetylated Hb).
Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
 Whole blood (EDTA, NaF-EDTA orheparin) is may be used
 For pretreatment of samples, available Ltype HbA1c Pretreatment Solution, which
is sold separately (manufactured bySEKISUI MEDICAL CO., LTD).
 Do not use highly hemolyzed whole blood.
2) Storage of samples.  Samples are stable for 7 days inrefrigeration.
 The stability of samples treated by L-type
HbA1c Pretreatment Solution (pretreatedsamples) is as follows.
Room temperature: 8 hours
Under refrigeration: 24 hours
Stored samples should be brought to room
temperature (15–30C) before use. Afterstorage for 2 hours or longer, thoroughly stir
pretreated samples before use.
2. Interfering substances
1) Assay results are not affected by free bilirubin(up to 50 mg/dL), conjugated bilirubin (up to50 mg/dL), formazin turbidity (up to 3000FTU), or ascorbic acid (up to 50 mg/dL).
2) Analytical results are not affected by modified
Hb (acetaldehyde-modified, acetylated, orcarbamylated)
obtainedbyadding
acetaldehyde, acetylsalicylic acid, or sodium
cyanate, respectively, to the sample at aconcentration of 50 mg/dL.
3) Analytical results are not affected by unstable
HbA1c obtained by adding glucose to thesample at a concentration of 1000 mg/dL.
4) Pay attention to measurement of samples

Description (Kit Components)
Component: IngredientsHbA1c Reagent 1: 10-(Carboxymethyl aminocarbonyl)-3,7-bis(dimethylamino) phenothiadine
sodiumHbA1c Reagent 2: Peroxidase
Fructosyl peptide oxidase
Intended Use
Measurement of hemoglobin A1c (HbA1c) in wholeblood
HbA1c is a glycation product of HbA0 accounting formost of hemoglobin (Hb). The HbA1c concentrationreflects the mean blood glucose level over a period of
1–2 months before testing, and it is used as an index ofglycemic control in patients with diabetes.1)
Assay Principle
1. Assay Principle
In the first reaction, glycosylated dipeptide derived
from the N-terminal of the β-chain of HbA1c is cut
out by a protease, and the Hb concentration issubsequently determined from the absorbance at aspecified wavelength. In the second reaction,hydrogen peroxide is produced by the action offructosyl peptide oxidase (FPOX) on glycosylated
dipeptide, and then causes color development by
10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino) phenothiazine sodium (coloringagent) in the presence of peroxidase (POD). TheHbA1c concentration is determined by measuring

1＊ ＊collected from patients dosed with aminoacid. In particular, do not use samples
containing a mixture of glucose and aminoacids, or falsely high values may be obtained.
3. Others
1) Always use L-type HbA1c Calibrator forcalibration.
2) Precautions for assay range
If the Hb concentration of the treated sample ishigher than the specified range, increase thevolume of pretreatment solution to twice thenormal volume (if the Hb concentration islower than the specified range, decrease thepretreatment solution volume to half of normal),and perform re-measurement. The Hbconcentration range varies among analyzers.
For example, with the Hitachi LABOSPECT
006 automated analyzer, re-measurement
should be performed if the Hb concentration
deviates from the range of 100–200 µmol/L.
Please contact SEKISUI MEDICAL CO., LTD.
to obtain the re-measurement ranges for otheranalyzers.

Assessment of Assay Results ＊1. Reference standard range1)3)
4.6–6.2 % (NGSP value)
2. There may be reactions or interfering reactions
with non-target substances. If assay results appear
to be unreliable, repeat the measurement (ifnecessary, after dilution) or try another analyticalmethods.
3. Some samples including extremely low activationof peroxidase (POD) may indicate the false high.
Therefore, assay results should be based on thepatient's clinical findings and other examination
results comprehensively.
Performance ＊1. Sensitivity
HbA1c: The absorbance is 0.050–0.100/min per
10 μmol/L of HbA1c.
Hb:
The absorbance is 0.10–0.30/min per 100
μmol/L of Hb.
2. Accuracy: 90–110% of the expected assay value
4. Within-run Reproducibility:
Coefficient of variation ≤ 5%(Test methods used for 1.–3. are in-housemethods.)
4. Measurement
Range4):(OnHitachi
LABOSPECT 006 automated analyzer)
HbA1c: 3.3–16.6% (NGSP value)(If the Hb concentration measured with a Hitachi
LABOSPECT 006 automated analyzer is 100–200µmol/L.)
5. Correlation4) (NGSP value)
N=197 r=0.995 y=1.00x–0.07
Control method: HPLC
6. Standard Material
Primary Reference Material for
Measurement of HbA1c (JCCRM411)

Dosage/Administration (Assay Procedure) ＊This product is used with the Hitachi LABOSPECT
automated analyzer. As an example, the procedure for
the Hitachi LABOSPECT 006 automated analyzer isshown below. Please contact SEKISUI MEDICALCO., LTD. to obtain information for other analyzers.
1. Preparation of reagents
Reagent (1): HbA1c Reagent 1 is ready to use.
Reagent (2): HbA1c Reagent 2 is ready to use.
2. Assay Procedure(1) Place 500 µL of L-type pretreatment solution
for NORUDIA N HbA1c (sold separately, andmanufactured by SEKISUI MEDICAL CO.,LTD.) in each test tube in advance.
(2) After centrifugation of the sample (800 g for 5minutes), collect 20 µL from the blood cell layer,
add it to the pretreatment solution mentioned inStep 1 above, and mix. Measure this mixture
with an automated analyzer as the pretreatedsample.
3. Assay Procedure

Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled asa material possibly infected with HIV, HBV,HCV, or other viruses. To prevent infection, use
disposable gloves and avoid mouth pipetting
during the test.
2) Sodium azide is added as an antiseptic agent in
the HbA1c Reagent 1. Therefore, if the reagent
comes in accidentally contact with eyes, mouthor skin, rinse immediately with ample water as
first aid, and consult the doctor if required.
2. Precautions for use
1) This product should be stored as directed,
without freezing. Freezing can deteriorate thereagents, which can produce inaccurate results.
Therefore, avoid using the reagents which havebeen previously frozen.
2) Black vial are used for HbA1c reagent 1 toprevent photodegradation. Avoid transfer intoother vial.
3) If the position of the cover of the automatedanalyzer (open/close) at the time of calibrationand measurement differs, accurate results may
not be obtained due to the difference in the levelof light entering the reaction cell. Close the coverof the automated analyzer during use.
4) Do not use expired reagents. Use of such

Pretreated
37°C
Reagent (1) ────▶Measurement
sample: +※1
75 μL
5 min (Absorbance )
15 µL
37°C
Reagent (2)Measurement
────▶
※2
20 μL
5 min (Absorbance )Calculate of────▶
concentration※1

Absorbance: The difference in absorbance between
480 nm and 800 nm and between 660 nm and 800nm※2
Absorbance: The difference in absorbance between
660 nm and 800 nm
Inter-item operation expressions for the automatedanalyzer are required for conversion to NGSPvalues (%).2HbA1c (NGSP%) = 91.5 × HbA1C (mol/L) / Hb(mol/L) + 2.15
Calibration material: L-type HbA1C Calibrator(Manufacture's assigned value)2

reagents cannot guarantee the reliability ofmeasurement values.
5) Do not replenish the reagents.
6) Do not perform the assay under direct sunlight
3. Precautions for Disposal
1) Before disposal, used samples and theircontainers must be immersed in sodium
hypochlorite solution at a concentration ofgreater than 0.1% for longer than 1 hour or
autoclaved at 121ºC for 20 minutes.
2 ）To prevent infections from spilled samples orsolutions containing samples, wipe the spilledarea thoroughly with disinfectants such as
sodium hypochlorite solution at a concentrationof greater than 0.1%.
3) The reagents and treated samples should bediscarded as medical waste or industrial waste
according to the waste disposal regulations.
4) The reagents should be disposed of in accordancewith the Water Pollution Control act or relatedregulations.
5) HbA1c Reagent 1 contains sodium azide. It canreact with lead or copper pipes to produce thehighly explosive metal azide. Therefore, thereagent should be flushed with large amounts ofwater when disposing.
4. Other precautions
Do not use the containers for other purposes.

ContactSEKISUI MEDICAL CO., LTD.
international@sekisui.com

Manufacturer ＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan

Storage and Shelf Life ＊Name
L typeNORUDIAN
HbA1c

HbA1cReagent
1HbA1cReagent
2Storagetemperature
2–10°C withprotection
from light
2–10°C

Shelf life

1 year fromthe date ofmanufacture(The expiration date is printed on the outer package.)
Packaging

L typeNORUDIAN HbA1c

Name
HbA1c Reagent(1)
1HbA1c Reagent(2)
2

Package2 × 35.3 mL
2 × 9.4 mL

OptionsName
L type Pretreatment Solution for
NORUDIA N HbA1c

Package2  33.9 mL

Shelf life: 1 year from the date of manufacture
References
1) Edited by the Japan Diabetes Society. [Treatment
Guide for Diabetes 2010]. Tokyo: Bunkodo; 2010.
p.9.
2) Wieland Hoelzel, Cas Weykamp, et al.: Clin Chem,
50, 166–174, 2004.
3) Atsunori Kashiwagi, et al.: Diabetol. Int. 3(1), 8–10, 2012.
4) In house data, SEKISUI MEDICAL CO., LTD.

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