CC 3-Laboratory:

Glucose Tolerance Test (GTT)

Blank Standard Sample

Distilled H20 10 uL
Glucose Standard 10 uL
Serum/Plasma 10 uL
Glucose Reagent 1 mL 1 mL 1 mL

 Incubate at 37 degree Celsius at heating bath for atleast 5 mins.

 Lactose load for adults: 50g of lactose dissolved in 400mL of H20

Conc. of Blood Glucose (mg%) = Absorbance of test x Conc. of standard

Absorbance of Standard

 Conversion factor: mg/dL to mmol/L : x 0.0556

 Expected Values: 70 – 105 mg/dL
 Final Color is stable for atleast 30 mins.

Procedure:
 After incubation, zero spectrophotometer with rgt blank.
 Read and record the absorbance of all tubes at 500 nm (wavelength)
 Final color is stable for atleast 30 mins.
 Concentration of the standard: 100 mg/dL
 Draw blood sample 30, 60, and 120 mins.
 1mL fasting blood sample in Sodium Fluoride anticoagulant
 Collect stool sample 5 hrs after giving lactose load. (Determine acidity and glucose content using strip method)
 Blood glucose method, use both strip and chemical method.

Laboratory Test Protocol:

o Normal Diet with High carbohydrates (150 g) for 3 days preceding the test.
o Fasting is required for 6-8 hrs before the test and not more than 16 hrs.
o Collect blood and urine samples and have the patient empty his bladder for each specimen.
o No food or drinks except water should be taken during the test periods.
o No alcohol should be taken the previous evening.
o No smoking allowed during test periods.
o Coffee and unusual physical exercise should be avoided for atleast 8 hrs before the test.

Principle:
Glucose Oxidase Oxidatively coupled w/
D-glucose D-gluconic acid , Hydrogen peroxide Red Quinoneimine dye
(Oxidation) 4-aminoantipyrine and Phenol
substituthbc, p-hydroxy-benzene
sulfonate(p-HBS) in the presence
of peroxidase

The amount of color red complex

is formed in proportional to
glucose concentration and
measured at 500-520 nm.