B Raghu MSc, PhD, FRSH
Asst. Professor of Biochemistry Asian Institute of Medicine, Science and Technology
Malaysia
Jitendar P Vij
Jaypee Brothers Medical Publishers (P) Ltd
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Practical Biochemistry for Medical Students
© 2003, B Raghu
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 author and the publisher.
First Edition : 2003
Publishing Director: RK Yadav
9788180611063
Typeset at JPBMP typesetting unit
Printed at Gopsons Papers Ltd, A-14, Sector 60, Noida
5PREFACE
This book is based on the curriculum for Medical undergraduates and covers a large number of basic experiments which help the Medical students understanding of theory and practice.
The theoretical introduction for each experiment, should give the students a solid foundation for understanding the practical aspects of Biochemistry.
This book is primarily intended for Medical students studying for professional examination in Biochemistry and similar examination. Dental undergraduates and student nurses will also find the book useful.
This book also contains eight useful case studies at the end of each clinical chemistry experiment.
I am grateful to Mr JP Vij, Mr RK Yadav, Mr Anil P Pillai, publishers for the cooperation extended during the presentation of book.
I thank my present and past students, colleagues, my wife Dr Vasundhara and my family for their patience during the preparation of the manuscript.
B Raghu
7SAFETY AWARENESS AND SAFE WORK PRACTICES IN THE LABORATORY
These are only guidelines. While they are not rules that will be stringently enforced, they should serve as a reminder of things you can do to work more safely. Although these guidelines are applicable to all research, teaching, and academic laboratories, your lab may require more specialized rules that apply to specific materials and equipment.
Laboratory Awareness
Label all storage areas, refrigerators, cabinets, etc., appropriately and keep all chemicals in properly labeled (noting date of receipt or generation and the date of the opening of the chemical) container(s).
Be alert to unsafe conditions and actions, and call attention to them so that corrections can be made as soon as possible.
Pour more concentrated solutions into less concentrated solutions to avoid violent reactions.
Be familiar with the appropriate measures you should take when you or someone in your lab is working with or is exposed to the following:
- Corrosive Chemicals
- Radioactive Materials
- Carcinogens
- Biohazards
- Compressed Gases
- Toxic Chemicals
- Reactive Chemicals
- Flammable Substances
PERSONAL SAFETY
Respiratory and Body Protection
- Use fume hoods whenever possible.
- Safety goggles/glasses with side shields should be worn at all times in the laboratory.
- Laboratory coat/apron should be worn in the laboratory.
- Gloves should be worn as needed.
Personal Hygiene
- Wash hands before leaving laboratory.
- Launder clothing worn in laboratory separately from other clothing.
- Never use your mouth to pipette chemicals.
- Avoid having long hair, loose sleeves/cuffs, rings, bracelets, etc. in close proximity to open flames or operating electrical machinery.
- Keep exposed skin covered. Shorts, skirts, or open-toed shoes should not be worn in the laboratory.
FIRE PREVENTION
Be aware of ignition sources in your laboratory area (open flames, heat, electrical equipment).
- Purchase and store flammable reagents in the smallest quantities possible. Do not store flammable liquids in standard refrigerators (an explosion-proof refrigerator should be used).
- Store flammable liquids in appropriate safety cabinets and/or safety cans.
- Do not store incompatible reagents together (e.g., acids with flammables).
- Do not store ethers for extended periods of time as explosive peroxides could form.
- Make sure that all electrical cords are in good condition.
WASTE COLLECTION
Minimize wastes at the source by limiting the quantities of materials purchased and used. Deposit all waste in designated containers. There are many different types of containers used for the collection of the various wastes. Be able to recognize these containers, and know which ones are appropriate for the wastes you generate.
WOUNDS
Small cuts and scratches
- Cleanse area with soap and water.
- Place a clean dressing over the wound.
Significant bleeding
- Calm and reassure the victim.
- Lay the victim down. This will reduce the chance of fainting.
- Do not remove any objects impaled in the victim.
- Put direct pressure on the wound with a sterile bandage or clean cloth.
- If direct pressure does not control the bleeding, elevate the wound above the heart if possible.
- If bleeding is severe elevate the victims legs about 12 inches, and cover the victim with a blanket.
- Do not apply torniquet
Thermal Burns
- First degree burns (e.g. sunburn or mild steam burn) are characterized by pain, redness and swelling.First aid procedures for first degree burns are as follows:
- Run cool water over the area of the burn or soak it in cool water for at least 5 minutes.
- Cover the burn with a sterile bandage or clean cloth.
- Do not apply any ointments, sprays or salves.
- Second and third degree burns are characterized by red or mottled skin with blisters (second degree), white or charred skin (third degree).
- First aid procedures for second and third degree burns are as follows:
- If the victim is on fire put the fire out.
- Do not remove any burnt clothing unless it comes off easily.
- Cover burns with dry sterile or clean bandaging.
- Do not apply any ointments, sprays or salves.
Chemical Burns
If hazardous chemicals should come into contact with the skin or eyes, follow the first aid procedures below.
- Do not become a victim, wear gloves and safety goggles to protect yourself if you are attempting to assist someone covered in chemical(s).
- Skin
- Remove victim's clothes — don't let modesty stand in the way.
- Remove victim's shoes — chemicals may collect in the shoes.
- Rinse the area with large quantities of water for at least 15 minutes (sink, shower or hose).
- Do not apply burn ointments/spray to affected areas.
- Cover with dry clean or sterile material.
- Eyes
- Eyelids have to be forcibly opened to ensure effective washing behind the eyelid.
- Be sure to wash from the nose out to the ear, this will avoid washing chemicals back into the eye or into an unaffected eye.
- Flood eyes and eyelids with water/eye solution for a minimum of 15 minutes.
- Remove contact lenses as soon as possible to rinse eyes of any harmful chemicals.
- Cover both of the victim's eyes with a clean or sterile gauze.
Ingestion of Chemicals
If the victim is awake and able to swallow, give water or milk. If they become nauseated, do not continue to administer fluids. If the victim is unconscious, turn their head or entire body onto their left side. Be prepared to start CPR, but be cautious about exposing yourself to chemical poisoning via mouth-to-mouth resuscitation. If available, use a mouth-to-mask resuscitator.
Chemical Spills
Acid Spills
- Apply neutralizer (or sodium bicarbonate) to perimeter of spill.
- Mix thoroughly until fizzing and evolution of gas ceases. NOTE: It may be necessary to add water to the mixture to complete the reaction. Neutralizer has a tendency to absorb acid before fully neutralizing it.
- Check mixture with Congo Red indicator paper. The red color indicates that the acid has been neutralized.
- Transfer the mixture to a plastic bag, tie shut, fill out a waste label, and place in a fume hood.
Solvent Spills
- Apply activated charcoal to the perimeter of the spill.
- Mix thoroughly until material is dry and no evidence of solvent remains.
- Transfer absorbed solvent to a plastic bag, tie shut, fill out and attach a waste label, and place in a fume hood.
Picture Hazard Warnings
- Picture hazard warnings help to identify the properties and classes of hazardous compounds. Examples include the flaming letter “O” (oxidizer), hand/bar of steel (corrosive) and a skull-and-crossbones (poison).
Biological Hazards
- Sources of biological hazards include bacteria, viruses, insects, plants, birds, animals, and humans. These sources can cause a variety of health effects ranging from skin irritation and allergies to infections (e.g. tuberculosis, AIDS), cancer and so on.
First Aid Kits
First aid kits should be standard equipment in every laboratory. Commercial, cabinet-type, or unit-type first aid kits are acceptable. A typical model for laboratories includes a variety of items specially selected to carry out emergency treatment of cuts, burns, eye injuries, or sudden illness. The first aid kit should contain individually sealed packages for each type of item. Contents of the kit should be checked weekly to ensure that expended items are replaced. Laboratory supervisors are responsible for maintaining the contents of the first aid kit.
No oral medication should be dispensed from the first aid kit.