UNIVERSITY OF MADRAS
B.Sc. DEGREE COURSE IN BIO-CHEMISTRY
SEMESTER SYSTEM WITH CREDITS
(Effective from the Academic Year 2003-2004)
SYLLABUS

Semester III - Paper III - Biophysical and Biochemical Techniques-I

Duration: 3 hrs
Maximum Marks: 100
Credits: 4

(90 hrs)

Unit I

Units of measurement of solutes in solution. Eg. Normality, molality, molarity and milliosmol. ionic strength. Examples of this concepts. pH, pOH, Henderson-Hesselbalch equation, buffers, pH of body fluids, buffers in body fluids, red blood cells and tissues. Measurement of pH by indicators, zwitter ions. pH dependent ionization of amino acids and proteins. Colloids and their application, viscosity, surface tension and Donnan membrane equilibrium. (18 hrs)

Unit II

ELECTROCHEMICAL TECHNIQUES:

Principles of electrochemical techniques. Measurement of pH by glass electrode and hydrogen electrode.
Oxygen electrode - principles, operation of a Clarke electrode, applications of oxYgen electrode. (18 hrs)

Unit III

CENTRIFUGATION TECHNIQUES:

Basic principles of centrifugation. Differential, density gradient, isopycnic and equilibrium centrifugation. Preparative and analytical ultracentrifugation techniques. Determinationof molecular weigth by centrifugation method (Derivation included). (18 hrs)

Unit IV

CHROMATO GRAPHIC TECHNIQUES:

General principles of chromatography. Principles, operational procedure and applications of
Paper chromatography
Thin layer chromatography
Ion exchange chromatography
Molecular sieve chromatography
Affinity chromatography
Gas Liquid chromatography
HPLC (l8 hrs)

Unit V

ELECTROPHORETIC TECHNIQUES:

General principles. Factors affecting the migration rate - sample, electric field, buffer and supporting medium.

Tiselius moving boundry electrophoresis.
PAGE
SDS- PAGE
Tmmunoelectrophrosis.
Cellulose acetate membrane electrophoresis
Agarose gel electrophoresis (18 hrs)

Books recommended

1. A biochemical Guide to principles and techniques of practical biochemistry - Keith Wilson and Kenneth H. Goulding
2. Principles and techniques of practical biochemistry - Bryan L. Williams and Keith Wilson
3. Practical Biochemistry - Shawney
4. Physical Biochemistry - David Friefelder
5. Instrumental methods of Analysis - Chatwal - Anand