Laboratory Manual for

Practical Biochemistry

Laboratory Manual for

Practical Biochemistry

Shivaraja Shankara YM
Associate Professor
Department of Biochemistry
KVG Medical College
Sullia, Karnataka
Co-Authors
Ganesh MK
Asst. Professor
Dept. of Biochemistry
KVG Dental College
Sullia, Karnataka

Hemavathi A
Asst. Professor
Dept. of Biochemistry
KVG Medical College
Sullia, Karnataka

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Laboratory Manual for Practical Biochemistry

2008, Jaypee Brothers Medical Publishers
All rights reserved. No part of this publication should be reproduced, stored in a retrieval
system, or transmitted in any form or by any means: electronic, mechanical, photocopying, recording, or otherwise, without the prior written permission of the authors and the
publisher.
This book has been published in good faith that the material provided by authors is
original. Every effort is made to ensure accuracy of material, but the publisher, printer
and authors will not be held responsible for any inadvertent error(s). In case of any
dispute, all legal matters are to be settled under Delhi jurisdiction only.

First Edition: 2008

ISBN 978-81-8448-259-1
Typeset at JPBMP typesetting unit
Printed at Rajkamal

Respectfully Dedicated To
Dr Kurunji Venkatramana Gowda
Founder President
Academy of Liberal Education
Sullia

Preface
Knowledge of practical aspects of biochemistry is essential for a
medical student to understand diagnostic and prognostic status of
any ailment. We have always felt the need for a complete and
comprehensive book on orientation towards various aspects of
practical biochemistry. This book is written in simple language,
covering the practical syllabus for MBBS, BDS, BSc MLT and BSc
nursing students as per the curriculum and guidelines of Rajiv
Gandhi University of Health Sciences, Bangalore and most of other
Health Universities in India. The book includes both theory and
practical aspects of clinical biochemistry. A clear-cut practical
concept has been offered in the book in respect of all qualitative
and quantitative experiments. Chapters on Demonstration
Experiments such as colorimetry, electrophoresis, chromatography,
etc. are also included. Questions at the end of each chapters and
normal values of various biochemical parameters in blood and
urine are also included in this book.
We hope that this book will meet the requirement of undergraduate students studying biochemistry. We will be glad to accept
constructive criticism and suggestions from the faculty, students
and readers to make this book a better one in subsequent prints.
Shivaraja Shankara YM
Ganesh MK
Hemavathi A

Acknowledgements
It is our privilege to express our sincere gratitude to
Dr KV Chidananda, Vice President and Dr KV Renuka Prasad,
General Secretary, AOLE for their constant encouragement and
support. We are grateful to Dr VK Jayapalan, Director, Dr KG
Nayak, Dean Academics, Dr Sheela G Nayak, Principal, KVG
Medical College and Hospital, Sullia for their constant inspiration
and encouragement. We also express our sincere thanks to
Mrs Shobha Chidananda, Treasurer, AOLE, Dr Jyothi R Prasad,
Director AOLE and Prof. AS Ramakrishna, Administrator, AOLE
Group of Institutions for their help and moral support. We
gratefully acknowledge the cooperation of Dr Cariappa, Asst. Prof.,
Department of Biochemistry and Mr Monappa and his office staff
in KVG Medical College.
We express our immense gratitude to Dr Shivashankara AR,
Associate Professor of Biochemistry, SDM Institute of Medical
Sciences, Dharwad for his timely help and valuable suggestions.
We are thankful to Mr Tarun Duneja, General Manager
(Publishing) and Mr K Suresh Kumar, Sales Manager, Jaypee
Brothers Medical Publishers (P) Ltd, New Delhi, for their sincere
effort and cooperation in bringing out this book in time.

Contents
QUALITATIVE EXPERIMENTS
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.

Carbohydrates
Reactions of carbohydrates
Scheme for identification of unknown carbohydrates
Proteins
Color reactions of proteins
Precipitation reactions of proteins
Scheme for identification of unknown proteins
Reactions of non-protein nitrogenous substances
Scheme for identification of
physiologically important substances
Urine analysis (Normal urine)
Analysis of abnormal constituents in urine
Analysis of gastric juice
Analysis of saliva

1
4
12
13
15
24
29
30
33
34
42
48
52

QUANTITATIVE EXPERIMENTS
14.
15.
16.
17.
18.
19.
20.
21.

Sample collection and preservation

Estimation of blood sugar by Folin-Wu method
Estimation of blood glucose by O-toluidine method
Estimation of blood urea by diacetyl
monoxime (DAM) method
Estimation of serum proteins and albumin/globulin
ratio by Biuret method
Estimation of serum albumin by Dye-binding method
Estimation of serum inorganic phosphate by
Fiske-Subba Row method
Estimation of serum cholesterol by Zaks method

55
58
63
65
69
73
75
79

xii Laboratory Manual for Practical Biochemistry

22. Estimation of serum uric acid by Caraway method
23. Estimation of serum transaminases by Reitaman
and Frankel method
24. Estimation of serum bilirubin by Malloy-Evelyn method
25. Estimation of serum creatinine by Jaffes alkaline
picrate method
26. Estimation of urine creatinine by Jaffes alkaline
picrate method

83
86
89
93
96

DEMONSTRATION EXPERIMENTS
27.
28.
29.
30.

Colorimetry general principle and procedure

Electrophoresis
Chromatography
Preparation of hemin crystals

99
104
109
113

ORAL GLUCOSE TOLERANCE TEST (ORAL GTT)

115
CASE REPORTS
121
NORMAL VALUES OF
BIOCHEMICAL PARAMETERS
129
Index

131