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Practical Biochemistry
Lecture 6:
Measurement of Blood Glucose
Prepared by: Dr. Slemani
M.Sc., Ph.D. in Clinical Biochemistry
E-mail: Asclinicalbiochemist@gmail.com

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Introduction
•Glucose is a monosaccharide.

• It is central molecule in carbohydrate

metabolism.

• Stored as glycogen in liver and

skeletal muscle.

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Entry of Glucose Into Cell

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Clinical Significance

• F.B.S (Fasting Blood Sugar)

• Fasting 12 - 14 hours

Normal Range= 70-99mg/dl

• Decrease
• Hypoglycemia
• Increase
• Hyperglycemia

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Hyperglycemia
Hyperglycemia refers to high levels of sugar, or glucose, in the blood.

Types of Diabetes:
❑ Diabetes Mellitus
- Insulin Dependent Diabetes Mellitus IDDM: type I
- Noninsulin Dependent Diabetes Mellitus (NIDDM) : type II
❑ Gestational Diabetes

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Insulin Dependent Diabetes Mellitus (IDDM)
• Absolute deficiency of insulin
• Make up about 10% of all patients with D.M.

• IDDM present at an early age (usually before 30)

• Clinical signs :
• Blood glucose

• Insulin (decreased)

• Ketosis

• Loss of body weight

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Noninsulin Dependent Diabetes Mellitus (NIDDM)
• Deficiency of insulin receptor

• Patients are commonly obese

• NIDDM is the most common from of D.M. ( 80-90%)

• Usually first present at an age over 40

• Clinical signs
• Blood glucose

• Insulin

• Osmotic diuresis

• Dehydration

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Diagnosis of Diabetes Mellitus

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Hypoglycemia
Hypoglycemia is a condition in which your blood sugar (glucose)
level is lower than normal.

Causes of hypoglycemia

• Insulinoma (islet cell tumors)

• Glycogen Storage Diseases (GSD)

• Addison’s disease

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 Blood Glucose Tests
• Fasting blood glucose test: This measures the amount of sugar in your blood.

• Glucose tolerance test: This measures your body's ability to use sugar after drinking a
standard amount in a sugary drink.

• 2HPP (two Hours Postprandial Plasma glucose):

• To screen for diabetes mellitus

• To monitor glucose control

• HA1C (Glycosylated Hemoglobin) test: This measures your average blood sugar level
over the last 2 to 3 months

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Methods Used
❑Enzymatic method:

❑Oxidation-Reduction methods

❑Condensation Methods

• Orto – Toluidine

❑Digital Glucometer

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O - Toluidine Method
➢ Principle
• Glucose react with Orthto-luidine in hot acidic medium to form a
Green color complex
• Color intensity α Concentration of Glucose
• Measured in photometer at 620 nm to 660 nm.
• It can measured other monosaccharide also.
• Ortho-toluidine is carcinogenic.

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Blood collection for glucose estimation :
• Samples:

• ✓ Blood samples

• --Whole blood

• --Serum

• --Plasma (with Ca. oxalates / NaF),

• ✓ Fresh urine

• ✓ CSF: collected in sterile clean container and to be done

immediately or

• centrifuged to get cell free fluid.

• ✓ Any other body fluid: to be treated like CSF

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Digital Glucometer
Glucometer (or glucose meter)
Glucometer is a medical tool that is used for measuring the approximate level
of glucose in the blood.

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Working principle of a Glucometer:
Most of the glucometers are based on electrochemical technology.
Glucometer test strips:
In each glucometer test strip which are to be used for determining glucose in the blood, contains an
enzyme called glucose oxidase. This enzyme then reacts with the glucose in the blood sample and
creates an acid called gluconic acid.
The gluconic acid thus formed then reacts with another chemical in the testing strip called ferricyanide.
The ferricyanide and the gluconic acid then combines with each other and forms ferrocyanide.
iii) As soon as the ferrocyanide has been formed the device (i.e., glucometer) runs an electronic
current through the blood sample on the strip.
This current thus generated is able to read the ferrocyanide and identify the amount of glucose
present in the blood sample on the testing strip.
That number is then displayed on the screen of the glucometer.

National Institute of Technology 2020-2021

 National Institute of Technology

National Institute of Technology 2020-2021