GLUCOSE DETERMINATION Flashcards
What are the 2 methods in glucose determination?
Enzymatic & Non-Enzymatic
What method is most common test in glucose determination, but is not the reference method?
Glucose Oxidase Method
What is the reference method for glucose determination?
Hexokinase method
What are the substrates for primary reaction?
B-D-Glucose and H2O2 + O2
What is the enzyme used for Primary reaction?
God - Glucose Oxidase
What are the products of primary reaction?
H2O2 (Hydrogen peroxide) and Gluconic acid
What are the color developers in secondary reaction?
4-aminoantipyrine + phenol
What are the substrates of Secondary reaction?
H2O2 + 4-aminoantipyrine + phenol
What is the enzyme for the secondary reaction?
POD - Peroxidase
What are the end products of the secondary reaction?
Quinonime complex + H2O
What quinone complex color formed at secondary reaction?
Red-violet
What is the relationship of the intensity of color to the glucose concentration?
Proportional
What is the purpose of phosphate buffer in the experiment?
To neutralize the pH level
What is the glucose standard, and wavelength?
100mg/dL ; 500 nm
The wavelength 500 NM is under what spectrum?
Visible spectrum
Enumerate the manual procedure of glucose oxidase method.
- Zero spectrophotometer at 500 NM with distilled water.
- For each standard and sample tubes, add 1.0 mL reagent to test tubes.
- Warm the tubes to 37C for 5 minutes
- Add 10 uL (0.010 mL) of SERUM in SAMPLE TUBE, while 10 uL (0.010 mL) of GLUCOSE STANDARD in STANDARD TUBE.
- Return to 37C incubation for 5 minutes.
- After 5 minutes of incubation, read and record the absorbance of all samples.
T or F: 37C = 99F = 310.15K
TRUE
What is the molecular formula of phenol?
C6H5OH
What is the molecular formula of 4-aminoantipyrine
C11H13N3O
What is the principle of result for glucose oxidase method?
To compare the Au with As that is identically treated.
What is the formula of glucose determination? With unit.
Glucose (mg/dL) = Au/As x 100
What is the reference range of serum/plasma, and its’ equivalence to mmol/L?
70-105 mg/dL ; 3.89 - 5.83 mmol/L
What is the reference range of CSF, and its’ equivalence to mmol/L?
40-75 mg/dL ; 2.22-4.17 mmol/L
Result: 70-105 mg/dL is read as?
Normal
Result: <70 mg/dL is read as?
Below normal
Result: >105 mg/dL is read as?
Above normal
