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Estimation of Glucose

The document outlines the procedure for estimating blood glucose levels using the Glucose Oxidase-Peroxidase (GOD-POD) method, detailing the principle, reagents required, and step-by-step procedure. It includes a calculation method for determining glucose concentration based on optical density measurements. Additionally, it provides self-learning questions related to glucose estimation and diabetes diagnosis.

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88 views2 pages

Estimation of Glucose

The document outlines the procedure for estimating blood glucose levels using the Glucose Oxidase-Peroxidase (GOD-POD) method, detailing the principle, reagents required, and step-by-step procedure. It includes a calculation method for determining glucose concentration based on optical density measurements. Additionally, it provides self-learning questions related to glucose estimation and diabetes diagnosis.

Uploaded by

alternatebeautyy
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Estimation of Blood Glucose

Competency: BI 11.21 Demonstrate the estimation of glucose, Creatinine, Urea and Total Protein in
serum

Aim : To estimate the glucose quantitatively in the given blood sample using Glucose
Oxidase -Peroxidase (GOD-POD) method

Principle : Glucose Oxidase oxidizes the specific substrate α-D Glucose to Gluconic acid
with generation of Hydrogen Peroxide H2O2. H2O2 thus produced is acted upon
by Peroxidase and Oxygen is liberated. The liberated oxygen is transferred to
chromogen system consisting of 4-aminoantipyrine and phenolic compound to
produced red quinoneimine dye. The intensity of the color is directly proportional
to the concentration of glucose and is measured colorimetrically at 505 nm (Green
Filter) against a blank and a standard.

Glucose Oxidase
Glucose + O2 + H2O Gluconate + H2O2

H2O2 + Phenolic Compound Peroxidase Quinoneimine complex


+ +
4-Aminoantipyrine H2O

Reagents Required:

1. Glucose (Enzyme/Chromogen)
2. Glucose Buffer-Phenolic Compound
3. Glucose Standard: Strength 100 mg/dL
4. Working reagent is prepared by gently mixing the enzyme/chromogen with buffer and
stored between 2o-8o C

Procedure:

Three test tubes are taken and labeled as Blank (B) Standard (S) and Test (T). The
reagents are dispensed in to each of these as follows:
Reagent Blank (B) Standard (S) Test (T)
mL mL mL
Working Reagent 1.0 1.0 1.0
Standard -- 10 µL --
Sample -- -- 10 µL
Distilled water 10 µL -- --

The contents are mixed well and the absorbance of Test (T) is measured against a
standard (S) and a blank (B) treated similarly.

Test Results:

Blank (B) Standard (S) Test (T)

O.D values

Calculation:

O.D T – O.D S

Glucose Concentration : ------------------------ X Concentration of Standard

(mg/dL) O.D T – O.D S

Report : The Blood glucose concentration in the given sample is _________


mg/dL

Self –Learning

1. Learn to convert the glucose value in mg/dL to mmol/L


2. Learn the Normal values of Fasting, Post-prandial and Random Blood Sugar values
3. List out the different methods –Chemical and enzymatic methods-for blood glucose
estimation
4. What is “True Glucose” value?
5. What is the preservative used to collect blood samples for glucose estimation?
6. What test is preferred to identify the sugars in urine sample?
7. What are the different criteria for diagnosis of Diabetes Mellitus?
8. What method is followed for Self Monitoring of Blood Glucose (SMBG) by Diabetic
patients?
9. What is HbA1c and what is its significance?
10. How do you classify Diabetes Mellitus?
11. What are the different complications of Diabetes Mellitus?

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