HbA1c REAGENT PREPARATION AND STABILITY: CALCULATION:

Reagents are all liquid ready to use. The analyser automatically calculates the HbA1c concentration in the sample.
HAEMOGLOBIN A1C Before use, mix reagent by gently inverting each bottle. Do not use the reagent described above for any other
purpose that described herein.
The calibrator values are aligned with the DCCT system and are therefore reported in the NGSP6 format.
IFCC values can be calculated using the following formula
MULTI PURPOSE This reagent kit is stable up to the expiry date stated on the label when stored at 2-8ºC.
For analysers capable of handling 3 reagents, R1A, R1B, R2 are ready to use. For analysers capable of
NGSP = [0.915 x IFCC] + 2.15 8,9

REAGENT handling only 2 reagents, HbA1c reagents R1A and R1B should be mixed in a 7:3 ratio and allowed to sit at 2-
8ºC for 24 hours prior to use. To prepare sufficient R1AB mixture, pour the entire contents of R1B into R1A
EXPECTED VALUES: 5
bottle. Mix gently by inversion. Normal 3.0 – 6.0% Controlled diabetics 6 – 9 %
KIT SPECIFICATIONS: Reconstituted R1AB thus prepared is stable for 4 weeks when stored at 2-8ºC. R1B and R2 reagents are light
Each laboratory should establish its own reference range. In using HbA1c to monitor diabetic patients, results
Cat. No. Quantity Reagent Storage sensitive.
should be interpreted individually.
1 x 18 ml R1A COMPONENT COMPOSITION:
1 x 7.5 ml R1B PERFORMANCE CHARACTERISTICS:
GL2201HC
1 x 11ml R2 Component Ingredients
Performance results can vary with the instrument used. Data obtained in each individual laboratory may
1 x 32 ml Lysis Buffer Reagent 1a MES Buffer, Proteases, Triton-X-100, Redox agents differ from these values.
2-8°C Reagent 1b MES Buffer, Redox agents
2 x 18 ml R1A Reagent 2 Tris Biuffer,FVO Enzyme, POD, Chromagen Linearity:

GL2211HC 2 x 7.5 ml R1B Lysis Buffer CHES Buffer, Triton-X-100 ,SDS, Redox reagents The assay range is 4.0% - 12.0%
2 x 11ml R2
TYPE OF SPECIMEN: Interfering substances:
2 x 32 ml Lysis Buffer Ascorbic Acid up to 12mg/dL
Bilirubin (Total) up to 15 mg/dl
Sample Collection: Use whole blood treated with EDTA anticoagulant. Bilirubin (Conjugate) up to 13 mg/dl
INTENDED USE: It is recommended to follow NCCLS procedures (or similar standardised conditions) regarding specimen Glucose up to 4000mg/dL
handling. Specimen should be collected in an appropriate sample container, with proper specimen Triglycerides up to 4000mg/dL
In Vitro Diagnostic reagent pack for the quantitative determination of Haemoglobin A1c (HbA1c in %) in whole identification.
human blood on automated and semi automated Clinical Chemistry Analyser Uric Acid up to 30mg/dL
Stability of HbA1C in whole blood: Up to 2 Weeks at 2-8ºC. Urea up to 80mg/dL
Acetylated, carbamylated and labile HbA1c do not adversely affect the Assay. Variant haemoglobin S, C and E
SUMMARY AND EXPLANATION: Sample preparation: do not significantly interfere with this assay
Glycosylated haemoglobin is formed by the combination of haemoglobin and glucose through a non-enzymatic Resuspend erythrocytes by gentle inversion(X5) prior to testing
Accuracy of the assay will be affected if whole blood is not thoroughly mixed prior to testing Precision:
reaction. The procedure is slow, permanent and irreversible, and lasts during the entire life cycle of a red blood
cell, (approximately 120 days). The rate of glycosylated haemoglobin synthesis is proportional to blood glucose Add 20µl of fully resuspended sample to 250 µl of Lysis Buffer. Mix gently using a pipettor without creating Within Run Mean SD % Between Run Mean SD %
concentrations.; therefore, glycosylated haemoglobin, measured as a percent of total haemoglobin, reflects foam. Incubate at 250C for 10 minutes until red blood cells are completely lysed. N=20 (%) CV N=5 (%) CV
the mean blood glucose concentration over the previous 1-3 months. This therefore provides a suitable marker Complete lyses is observed when the mixture becomes a clear dark red solution without any particle matter Level 1 5.7 0.06 1 Level 1 5.7 0.10 1.8
for the assessment of long term glycaemia in diabetic patients. Incubate the sample for longer if necessary, to obtain complete lyses Level 2 10.3 0.07 0.7 Level 2 10.3 0.18 1.8
The lysate will be stable for up to 4 hours at room temperature. Method Comparison:
Calibrators and controls should be treated the same as patient samples prior to use. A comparison with another commercially available HbA1c method gave the following results
PRINCIPLE OF THE TEST:
Glenbio Direct Enzymatic HbA1c assay directly determines the % HbA1c in the sample and does not require Y = 1.02 + 0.0135, r2 = 0.9874
an additional measurement of total haemoglobin. Samples are lysed and reacted with agents to eliminate TEST PROCEDURE: 44 samples were tested in the range 5 – 13%
signal interfering substances. The lysed whole blood then undergoes protease digestion, a process which
Materials required but not supplied: BIBLIOGRAPHY:
releases amino acids from the haemoglobin beta chains. Glycated valines released in this process serve as
1. Goldstein DE et al Diabeter Care 27(7): 1761-73 (2004)
substrates for specific recombinant fructosyl valine oxidase (FVO) enzyme. FVO specifically cleaves N-
Description Catalog. No. Description Catalog. No. 2. United Kingdom Prospective study, Lancet 352: 837 – 53 (1998)
terminal valines, producing hydrogen peroxide, the rate of production of which can be measured
HbA1c Calibrator set GL9708 Photometer N/A
3. The Diabetes Control and Complications Trial Research Group
spectrophotometrically. 4. Little R et al Clin Chemistry, 47:1985 – 1992 (2001)
HbA1c Control Level Set GL9104 General Laboratory Equipment N/A 5. American Diabetes Association. Clinical Practice Recommendation; standards of medical care for patients with diabetes mellitus
6. NGSP http://www.missouri.edu/~doabetes/ngsp.hmtl
WARNINGS AND PRECAUTIONS: Assay procedure: 7. Goldstein et al, Clin Chem 32: B64 – B70 (1986)
For In Vitro Diagnostics Use Only - For Professional Use Only Wavelength: λ: 700 nm
8. Hoelzel W et al. IFCC reference system for measurement of haemoglobin A1c in human blood and the national standardisation
Carefully read instructions for use. Deviations from this procedure may alter performance of the assay. schemes in the USA, Japan and Sweden: a method-comparison study. Clin Chem 2004;50:166-74
Temperature: 37°C 9. Sacks, D (ed) Global Harmonization of Haemoglobin A1c. Clin Chem 51 (4): 681 – 683 (2005)
Optical path: 1 cm light path.
Components Colour and Appearance:
SYMBOLS:
Reagent 1a: Clear solution.
Calibrator Sample The following symbols are used in the labelling of Glenbio systems:
Reagent 1b: Pale yellow solution
Reagent 2: Pale yellow solution. Reagent 1A 180 µl 180 µl IVD REF
Haemolysis solution: Clear colourless liquid. Reagent 1B 75 µl 75 µl In Vitro Diagnostics Catalogue No
Any significant changes could indicate that the assay might be compromised. Refer to Laboratory’s QC Sample 40 µl LOT CONT
Batch Code Content
program for actions to be taken. In case of serious damage to the bottle and/or cap, resulting in product Calibrator 40 µl
leakage and/or contamination, do not use the reagent pack and contact your distributor. Gently mix and Incubate at 37°C Measure the Optical Density (A1) after 4 minutes. REAG Reagent Ab Antibody
Reagent 2 110µl 110 µl
Safety precautions: CAL Calibrator SUBS Substrate
Measure the Optical Density (A2) after 3 minutes. Calculate ∆Abs= A2-A1
CAUTION: Take all necessary precautions required when handling laboratory reagents. Product is not
hazardous under EU specifications. BUF
Use the calibration curve constructed to obtain % HbA1C from absorbance calculated Buffer CE Mark - Device complies with the Directives 98/79/EC
Handling precautions:

Take the necessary precautions required for handling all laboratory reagents. Calibration: Storage temperature → Reconstitute with

Do not use components past the expiry date stated on the Bottles. Using recommended Calibrators, calibrate the assay:

Do not use components for any purpose other than described in the “Intended Use” section.
Daily Expiry Date Manufactured By

Avoid contact with eyes and skin. If contacted, flush eyes and risen skin with a large amount of water. If
When using a new reagent kit or changing lot number. (Last day of the month)
irritation persists, consult with a physician.
Following preventive maintenance or replacement of a critical part of the photometer used.

Do not interchange caps among components as contamination may occur and compromise test results.
When Quality Control results are out of range. Biological risk Consult Instruction for Use

Refer to local legal requirements for safe waste disposal.
Quality Control: GLENBIO LTD
INSTRUMENTS: All clinical laboratories should establish an Internal Quality Control program. Verify instrument and reagent 10 Kilbegs Road, Antrim, Co. Antrim, BT41 4NN
Instrument Applications are available on request performance with recommended controls or similar. The values obtained for QC should fall within Tel/Fax: +44(0)2879659842
manufacturer’s acceptable ranges or should be established according to the Laboratory’s QC program:
Email: [email protected]
Controls should be assayed:
Web: www.glenbio.com

Prior reporting patient results.

Following any maintenance procedure on the photometer used.

At intervals established by the Laboratory QC Programme. GLENBIO IRELAND LTD
17b Fota Business Park, Carrigtwohill, Co. Cork,
T45 PK77, Ireland

Page 1 of 1 Revision: 03 Issued on: 22 July 2021