This experiment has to be written in the quantitative portion allotted in the record notebook.
If you guys have any doubt contact me (Dr.Swathika)
This is to be written on the right side of the record notebook
ESTIMATION OF BLOOD GLUCOSE BY GOD-POD METHOD
GOD-POD method is an enzymatic method based on oxidation-peroxidation reaction, used for
the estimation of blood glucose.
PRINCIPLE:
Glucose is oxidized by Glucose oxidase to gluconic acid and hydrogen peroxide. In the presence
of enzyme peroxidase, released hydrogen peroxide is coupled with phenol and 4 –
aminoantipyrine(4-AAP) to form a pink coloured quinoeimine dye. Absorbance of the coloured
dye is measured at 505 nm and is directly proportional to glucose concentration in the sample.
PROCEDURE:
Sample : Whole blood or plasma (Grey coloured stopper vacutainer tube which has the
additives potassium oxalate and sodium fluoride in the ratio of 3:1. Potassium oxalate is an
anticoagulant and sodium fluoride inhibits enolase enzyme to prevent glycolysis.)
Step 1: OXIDATION – PEROXIDATION:
REAGENTS BLANK(B) STANDARD(S) TEST(T)
Distilled water 0.01 ml - -
Plasma(sample) - - 0.01 ml
Glucose standard - 0.01 ml -
Glucose reagent 1 ml 1 ml 1 ml
Mix well and incubate at room temperature for 10 minutes. Read the colour intensities on
colorimeter at 505 nm wavelength.
Step 2:DATA COLLECTION AND CALCULATION:
1. ‘T’ OD
2. ‘S’ OD
3. Strength of standard = 0.01 mg%
4. Volume of sample = 0.01 ml
Blood Glucose concentration = Test(T) OD x Conc. Of Standard x 100
Standard (S) OD Volume of sample
RESULT:
The concentration of glucose in the given sample is ___________ mg/dl.
Plasma glucose reference ranges
Fasting plasma glucose: < 110 mg/dL
Postprandial plasma glucose: < 140 mg/dL (2 hrs)
Random plasma glucose: <140 mg/dL
Hypoglycemia (reduced blood glucose concentration)
Plasma Glucose level of < 50 mg/dL is said to be hypoglycemia.
Common causes include fasting, endocrinological disorders (insulinoma, hypothyroidism,
hypopituitarism, Addison’s disease), enzyme deficiencies etc.
Hyperglycemia (elevated blood glucose concentration)
Common causes include type-1 & type 2 diabetes mellitus, drugs (steroids, beta blockers),
endocrinological disorders (hyperthyroidism, hyperpituitarism, Cushing’s disease, glucagonoma,
and pheochromocytoma).
Diabetes mellitus (DM)
Diabetes mellitus is a metabolic disease due to absolute or relative insulin deficiency.
Types:
Type 1 DM
Type 2 DM
Gestational DM, Impaired glucose tolerance (IGT), Impaired fasting glucose (IFG)
Biochemical investigations in Diabetes
Blood glucose level, lipid profile, kidney functions, glycated hemoglobin, fructosamine.
WHO criteria for the diagnosis of Diabetes Mellitus
Symptoms of diabetes plus random blood glucose concentration ≥ 200 mg/dL or
Fasting plasma glucose ≥ 126 mg/dL or
Two-hour post-load plasma glucose ≥ 200 mg/dL during OGTT
Fasting plasma glucose between 110–125 mg/dL is impaired fasting glucose.
Fasting plasma glucose between 110–125 mg/dL and 2 hour after oral glucose load between
140 and 199 mg/dL is impaired glucose tolerance.
