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 (10th edition)* Revised: January2017 (9th edition)

This package insert must be read carefully prior to use.
Glycohemoglobin A1c assay kit(Classification No.: 30168000)

concentration is determined by measuring theabsorbance of this complex.
The percentage of total Hb existing as HbA1c is
calculated from the concentrations of HbA1c andHb.

NORUDIA N HbA1c
General Precautions
1. This product is for in vitro diagnostic use, andmust not 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 beguaranteed 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 ifrequired.
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.

HbA1c:Protease
HbA1c ────▶ Glycosylated dipeptide
FPOX
Glycosylated dipeptide + O2 + H2O ────▶
Glucosone + Dipeptide + H2O2POD
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) Applicable to various automated analyzers.
5) Analytical results are not affected by unstable
HbA1c or modified Hb (such as carbamylatedor acetylated Hb).
Procedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
 Whole blood (EDTA, NaF-EDTA or
heparin) may be used.
 For pretreatment of samples, use
Pretreatment Solution for NORUDIA N
HbA1C, which is sold separately(manufactured by SEKISUI MEDICALCO., 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
pretreatment solution for NORUDIA N
HbA1c (pretreated samples) 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)
obtainedbyaddingacetaldehyde, acetylsalicylic acid, or sodium
cyanate, respectively, to the sample at aconcentration of 50 mg/dL.
3) Analytical results are not affected by unstable

Description (Kit Components)
Component: Ingredients
HbA1c Reagent 1: 10-(Carboxymethyl amino
carbonyl)-3,7-bis(dimethylamino) phenothiadine
sodiumHbA1c Reagent 2: Peroxidase
Fructosyl peptide oxidase
Intended Use
Measurement of hemoglobin A1c (HbA1c) inwhole blood
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
of glycemic 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 ofHbA1c is cut out by a protease, and the Hbconcentration is subsequently determined fromthe absorbance at a specified wavelength. In thesecond reaction, hydrogen peroxide is producedby the action of fructosyl peptide oxidase (FPOX)
on glycosylated dipeptide, and then causes color
developmentby
10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino)
phenothiazine sodium (coloring agent) in thepresence of peroxidase (POD). The HbA1c1

HbA1c obtained by adding glucose to thesample at a concentration of 1000 mg/dL.
4) Pay attention to measurement of samples
collected from patients dosed with aminoacid. In particular, do not use samples
containing a mixture of glucose and aminoacids, or falsely high values may beobtained.
3. Others
1) Always use HbA1c Calibrator for NORUDIA
N HbA1c for calibration.
2) Precautions for assay range
If the Hb concentration of the treated sample
is greater than 310 μmol/L, increase thevolume of pretreatment solution to twice thenormal volume (if the Hb concentration is 90
μmol/L or lower, decrease the pretreatment
solution volume to half of normal), andperform re-measurement.

3.

with non-target substances. If assay resultsappear to be unreliable, repeat the measurement(if necessary, after dilution) or try anotheranalytical methods.
As there is a risk of assay results being
artificially high in a sub-portion of samples withextremely low peroxidase (POD)-like activity,
physicians
shouldassessallresults
comprehensively in conjunction with clinicalsymptoms and other findings.

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 100μmol/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): (On Hitachi 7170
automated analyzer)
HbA1c: 3.3–16.6 % (NGSP value)(If the Hb concentration is 90–310 µmol/L.)
5. Correlation4) (JDS values)
N=137
r=0.991 y=0.98x+0.04
Control method: HPLC
6. Standard Material
Primary Reference Material for Measurement ofHbA1c (JCCRM411)

Dosage/Administration (Assay Procedure) ＊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 Pretreatment Solution for
NORUDIA N HbA1c (sold separately) in eachtest tube in advance.
(2) After centrifugation of the sample (800 g for 5minutes), collect 25 µL from the blood cell
layer, add it to the pretreatment solution
mentioned in Step 1 above, and mix. Measure
this mixture with an automated analyzer as thepretreated sample.
3. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.

Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled
as a 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 inthe 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 photo-degradation. Avoid transfer into
other 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 thecover of the automated analyzer during use.
4) Do not use expired reagents. Use of suchreagents 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

37C
PretreatedMeasurement
Reagent (1)sample +────▶
180 μL(Absorbance※1)
12 μL
5 min37CMeasurement
Reagent (2)
────▶
60 μL
(Absorbance※2)
5 min
Calculate of───▶
concentration※ 1

Absorbance: The difference in absorbance
between 480 nm and 800 nm and between 660 nmand 800 nm※ 2
Absorbance: The difference in absorbance
between 660 nm and 800 nm
Inter-item operation expressions for theautomated analyzer are required for conversion toNGSP values (%).2)
HbA1c (NGSP%) = 91.5 × HbA1c (μmol/L) / Hb(μmol/L) + 2.15
Calibration material: HbA1c calibrator for
NORUDIA N HbA1c (Manufacture's assignedvalue)＊ ＊Assessment of Assay Results ＊1. Reference standard range1)3)
4.6–6.2% (NGSP value)
2. There may be reactions or interfering reactions

2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 inaccordance with the Water Pollution Control act
or related regulations.
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.

international@sekisui.comManufacturer ＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan

Storage and Shelf Life ＊NameNORUDIAN HbA1c

HbA1cReagent 1
HbA1cReagent 2

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

Shelf life

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

NORUDIAN HbA1c

Name(1) HbA1c Reagent 1Package
2 × 30 mL(2)

2 × 10 mL

HbA1c Reagent 2

Constituent reagents are available in otherconfigurations. For further details please contact
SEKISUI MEDICAL CO., LTD.
OptionsName
HbA1c Pretreatment Solution forNORDIA N HbA1cPackage
2 × 200 mL

Shelf life: 1 year from the date of manufactureThis product is also available in different packages.
Please contact SEKISUI MEDICAL CO., LTD.
References
1) Edited by the Japan Diabetes Society: Treatment
Guide for Diabetes 2010, 9, Bunkodo, 2010.
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.
ContactSEKISUI MEDICAL CO., LTD.

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