Purpose of the Test
To determine the concentration of glucose in serum samples.
Principle
Glucose is oxidized by glucose oxidase to form gluconic acid and hydrogen peroxide. The hydrogen peroxide reacts with phenol and 4-aminophenazone in the presence of peroxidase to form a red quinoneimine dye. The intensity of the color formed is proportional to the glucose concentration in the sample.
Equipments
1. Semi-autoanalyzer
2. Cuvettes
3. Micropipettes
4. Timer
5. Spectrophotometer
6. Glucose Reagent (Mono Reagent)
Sample Type
Serum or plasma is acceptable for the Test
Procedure
1. in 3 clean dry tubes add reagents and sample as mentioned in the Table below
| Blank | Standard | Test | |
|---|---|---|---|
| Glucose Reagent | 1000 μl | 1000 μl | 1000 μl |
| STD | – | 10 μl | – |
| Sample | – | – | 10 μl |
Procedure
2. Mix Well and incubate for 10 mins at room Tempreature or 5 min @ 37c’
3. using the colorimeter or spectrophotometer read the absorbance against blank at filter wavelenght 540 nm
4. then do the calculations based on the following formula
Glucose = Absorbance of test / Absorbance of standard x Conc of standard
The STD Conc is 100 mg/dl or may vary depending on the reagent brand.
Normal Range
Quality Control:
1. Run two levels of control with each batch of samples.
2. Results should fall within established confidence limits.
3. Review QC results and take corrective action if needed.
Warning :
1. Hemolysis causes falsely decreased results.
M. Sulieman
mohammad@mlsgaate.com
Check glucose reagent expiration. Inspect instrument for errors. Run controls before testing samples.
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