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The document outlines the importance of glucose in human life, detailing its metabolism and regulation through hormones like glucagon and insulin. It describes methods for blood glucose monitoring, glucose tolerance tests, and the enzymatic principle behind the glucose oxidase method for estimating glucose levels in serum. The methodology section provides a specific experimental setup for measuring glucose concentration in samples.

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36 views

Assessment 1

The document outlines the importance of glucose in human life, detailing its metabolism and regulation through hormones like glucagon and insulin. It describes methods for blood glucose monitoring, glucose tolerance tests, and the enzymatic principle behind the glucose oxidase method for estimating glucose levels in serum. The methodology section provides a specific experimental setup for measuring glucose concentration in samples.

Uploaded by

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

(Glucose Oxidase method)


INTRODUCTION
A constant source of blood glucose is an absolute requirement for human life. Glucose is the greatly
preferred energy source for the brain. Most of the carbohydrate consumed is absorbed as glucose. This glucose
is converted to glycogen (glycogenesis). Surplus amounts are also stored as fat in adipose tissues. In a normal
person, blood contains 60-90mg of glucose in the fasting conditions (post-absorptive state, 12-14 hours after the
last meal) and 110-140mg following meal, per 100ml (= 1 dL) of blood. In the fasting state, glucose is supplied
by the breakdown of glycogen (glycogenolysis). Liver can also produce glucose from non-carbohydrate source
(aminoacids), called gluconeogenesis. During periods of prolonged starvation, when no carbohydrate is
available from food, this process takes place.
When there is low blood glucose (due to heavy exercise, starvation), glucagon hormone is secreted by α-cells of
pancreas. Liver releases glucose into blood through glycogenolysis and gluconeogenesis. When there is high
blood glucose level, insulin hormone is secreted by β-cells of pancreas. Insulin lowers blood sugar level by
removal of glucose from blood. A balance between the two process of addition and removal of glucose from
blood maintains homeostasis of blood glucose.
Blood glucose monitoring:
 Chemical test strips
 Glucometers (accu-check)
 Continuous glucose monitoring using sensors under the skin
 Glucose sensing bio-implants
 Non-invasive methods (near IR, Ultrasound, dielectric spectroscopy)
Glucose tolerance test:
Glucose is given orally to determine how quickly it is cleared from the blood. This has been done to check for
diabetes. Patient should fast for 8-14 hrs, before measuring the glucose level. Then glucose has been given
orally to drink. The standard dose is 1.75g glucose/kg of body weight. Maximum dose should not exceed 75g.
Tests are done every 2 hrs for at least 5-times to determine whether the patient is diabetic or not. In the case of
diabetic patients, the fasting glucose level will be more than 126mg/dL and the level after 2 hrs will be more
than 200mg/dL.
Decrease of glucose tolerance: This occurs in diabetes patients. Blood sugar will rise above 180mg/dl, the renal
threshold for glucose, and glycosuria occurs (excretion of glucose in urine).
Increased glucose tolerance: This is due to overproduction of insulin, results in hypoglycemia (low blood sugar)
(<70mg/dL).

AIM
Estimation of glucose in serum

PRINCIPLE
Glucose is converted to gluconic acid by the enzyme glucose oxidase (GOD). Hydrogen peroxide is
formed in the reaction and this is converted to water and molecular oxygen by the enzyme peroxidase. The
released oxygen combines with the oxygen acceptor, 4-amino-phenazone, together with phenol to form a pink
colour, the intensity of which is measured at 515nm or with a green filter.

Glucose Oxidase
C6H12O6 + O2 + H2O C6H12O7 + 2H2O2

Catalase
2H2O2 2H2O + O2

METHODOLOGY
Ingredients Blank (B) Standard (S) Test(T)
Glucose oxidase 1.0ml 1.0ml 1.0ml
(GOD)
Distilled water 0.01ml - -
Glucose solution - 0.01ml -
Sample solution - - 0.01ml

OBSERVATION
Note down the lower limit and upper limit of glucose sample and identify whether it falls under the
normal range.

RESULT
Low limit: 70mg/dL
High limit: 160mg/dL
Quantity of glucose in the given sample --------------------
Thus the patient suffers from --------------------------

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