Bring reagents at Room Temperature before use.
Safety precautions
For in vitro diagnostic use only.
Do not pipette by mouth.
Exercise the normal precautions required for
handling laboratory
reagents.
Glucose
Enzymatic Colorimetric Method
principle
Enzymatic colorimetric determination of Glucose
according to the following reactions:
GOD
Glucose + O2
Gluconic Acid + H2O2
POD
2 H2O2 + Phenol + 4 - Aminoantipyrine
Red Quinone + 4 H2O
Intensity of the colour is direct proportional to the
Glucose concentration in the sample
*Reagents composition
Reagent 1
Glucose standard
100 mg/dl
Reagent 2
Phosphate Buffer pH 7.0 100 mmol/l
Phenol
10 mmol/l
GOD (Glucose Oxidase) 10000 U/l
POD (Peroxidase)
1000 U/l
4 - Aminoantipyrine
0.3 mmol/l
Sodium Azide
< 0.1%
According to the present laws the kit does not
contain substances classified as dangerous
*Procedure
1. Wavelength
Hg 510 nm (500 - 550)
2. Cuvette light path:
1 cm
3. Temperature
37°C
4. Adjust the instrument to zero against air or
distilled water
Pipette into cuvettes Blank Sample standard
Distilled water
10 µl
Sample
10 µl
standard
10 µl
Reagent
1 ml
1 ml
1 ml
Mix, incubate for 10 minutes at 37°C or 15 minutes
at Room Temperature.
Read the Absorbance of Calibrator and Samples
against the Blank at Wavelength of 510 nm.
Final color is stable at least 30 minutes
*Calculation
Serum, plasma and CSF
A Sample
Glucose (mg/dl) = ------------ x standard Value
A standard
Urine
A Sample
Glucose (mg/dl) = ------------ x standard Value x 10
A standard
Storage and Stability of Reagents
Store the kit at 2 - 8°C
All the Components are stable until the stated
expiration date if stored tightly closed and
refrigerated.
Urine/24 h
Preparation and Stability of Working solution
Reagent liquid and ready to use
Calibrator liquid and ready to use
Once opened the bottle, the reagent is stable for at
least 30 days if stored tightly closed, refrigerated and
protected from light
Serum, plasma:
CSF:
Urine:
Glucose in urine (mg/dl)
Glucose (g/24 h) = -------------------------------- x Urine Volume 24 h (lt)
100
Each laboratory should establish its Normal
Reference Range
Performance Characterics
A. LINEARITY LIMIT
The reaction is linear up to 500 mg/dl
For higher value dilute sample 1:2 with normal
saline, repeat the test and multiply the value by 2
B. DETECTION LIMIT
Values less than 3 mg/dl give non - reproducible
results
C. INTRA - ASSAY PRECISION (N = 20 replicates)
Mean (mg/dl)
SD
CV%
Control 1
98
3.42
3.49
Control 2
247
6.49
2.63
D. INTER - ASSAY PRECISION (20 replicates for 3
days)
Mean (mg/dl)
SD
CV%
Control 1
98.4
2.34
2.38
Control 2
245.7
4.35
1.76
E. ACCURACY
Comparation between this method (y) and another
commercial one (x), gave the following results:
N = 20 r = 0.986085 y = 1x - 5.5
F. INTERFERENCES
1. Haemoglobin up to 400 mg/dl does not interfere
2. Bilirubin up to 15 mg/dl does not interfere
3. Triglycerides up to 1000 mg/dl do not interfere
4. Ascorbic Acid up to 30 mg/dl does not interfere
Quality Control
Control sera are recommended to monitor the
performance of manual and automated assay
procedures.
Bibliography
1. Henry R. J.: Clin. Chem, Hoeber N. Y. 625 (1964)
2. Trinder P.: Ann. Clin. Biochem 6, 24 (1969)
3. Sharp P.: Clin. Chem Acta 40, 115 (1972)
For in vitro diagnostic use only.
The following symbols are used on labels
For in vitro diagnostic use
Reference values
Use by (last day of the month)
60 - 110 mg/dl
50 - 70 mg/dl
< 0.5 g/24 h
These values should only be used as a guideline.
Temperature limitation
Batch Code
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