Biomolecular and Bioanalytical Techniques

Theory, Methodology and Applications

Vasudevan Ramesh, Vasudevan Ramesh

• Publisher: Wiley
• Year: 2019
• ISBN:
  9781119484011
• Edition: 1
• Topic: Physical Sciences
• Subtopic: Analytic Chemistry

An essential guide to biomolecular and bioanalytical techniques and their applications

Biomolecular and Bioanalytical Techniques offers an introduction to, and a basic understanding of, a wide range of biophysical techniques. The text takes an interdisciplinary approach with contributions from a panel of distinguished experts. With a focus on research, the text comprehensively covers a broad selection of topics drawn from contemporary research in the fields of chemistry and biology. Each of the internationally reputed authors has contributed a single chapter on a specific technique. The chapters cover the specific technique's background, theory, principles, technique, methodology, protocol and applications.

The text explores the use of a variety of analytical tools to characterise biological samples. The contributors explain how to identify and quantify biochemically important molecules, including small molecules as well as biological macromolecules such as enzymes, antibodies, proteins, peptides and nucleic acids. This book is filled with essential knowledge and explores the skills needed to carry out the research and development roles in academic and industrial laboratories.

• A technique-focused book that bridges the gap between an introductory text and a book on advanced research methods
• Provides the necessary background and skills needed to advance the research methods
• Featuresa structured approach withineach chapter
• Demonstrates an interdisciplinary approach that serves to develop independent thinking

Written for students in chemistry, biological, medical, pharmaceutical, forensic and biophysical sciences, Biomolecular and Bioanalytical Techniques is an in-depth review of the most current biomolecular and bioanalytical techniques in the field.

1
Principles of Health and Safety and Good Laboratory Practice

Elaine Armstrong

Health & Safety Services, Compliance & Risk, University of Manchester, Oxford Road, Manchester, M13 9PL, UK

1.1 Introduction

Scientific research, by definition, involves carrying out novel work to further scientific knowledge and, in pursuit of this activity, new techniques are developed and applied. The object of this chapter is to discuss a set of principles and guidelines that, when followed, will provide a safe and healthy environment for researchers, which will, in turn, facilitate and promote good science.

Chemical and biological laboratories are potentially very hazardous places in which to work. In recent years there have been a number of very serious accidents in academic laboratories with tragic and sometimes fatal consequences for those involved. These include fatalities in 2009 when a researcher died of extensive burns due to contact with a pyrophoric chemical [1] and in 2011 when a researcher was asphyxiated in an oxygen depleted atmosphere caused by evaporation of liquid nitrogen into a non‐ventilated space [2]. Serious injuries were caused to a graduate student in 2010 who was grinding energetic material that exploded [3] and a researcher lost an arm when a pressure vessel exploded in 2016 [4].

The risk of accidents and injury can be significantly reduced by researchers being aware of any potential hazards and working with care and attention to detail. Prior to commencing any work activity, it is very important, and time well spent, for researchers to familiarise themselves with all available information about the materials, equipment and processes that they will be using during the course of their work. The safety of everyone in the laboratory is largely determined by each individual's work practices.

1.2 Good Laboratory Practice

Good Laboratory Practice, or GLP, is a series of behaviours that is designed to prevent accidents, many of which will be described in the specific procedures developed by administrators or principal investigators for use in their laboratories. However, some general guidelines are given below:

• Do not eat, drink, smoke or apply cosmetics in the laboratory.
• Wash and dry hands before leaving the laboratory.
• Wear shoes with a closed toe – no sandals or flip flops.
• Wear personal protective equipment (PPE) that is required by the relevant risk assessment, properly (safety spectacles worn on top of the head do a poor job protecting eyes from chemical splashes).
• Cover any broken skin with suitable dressings.
• Keep benches and fume cupboards clear of unnecessary equipment, which leaves room for carrying out the work and will minimise the effect of any accidents.
• Ensure that all chemicals are properly labelled with the name of the chemical and any hazard information and, for samples, the owner's name, date of preparation and quantity.
• Replace lids and stoppers.
• Return chemicals to their dedicated storage areas after use.
• Check chemical stock and equipment that is not in regular use periodically to ensure it is in good condition and specific storage conditions are being met (e.g. certain chemicals should not be allowed to ‘dry out’).
• Store chemicals safely in appropriate storage spaces.
• When carrying large bottles of solvent, always use suitable carriers and do not lift large bottles solely by the neck.
• Keep substances that are incompatible with each other apart and in separate storage spaces, and label them clearly.
• Comply with local restrictions on the amount of highly flammable and flammable materials (which includes waste).
• If equipment becomes faulty, take it out of service, label it and report it to someone who will arrange for its repair.
• Use all equipment in accordance with the manufacturer's instructions.
• Dispose of all out of date and/or unwanted chemicals and equipment safely, on a timely basis and according to local procedures.
• Inspect any glassware before use and do not use any that is broken, chipped or cracked, as this might either directly cause injury to the researcher or fail catastrophically in use.
• Follow any local rules and guidance about working alone.
• Follow any local rules and guidance about working out of hours.

In addition to using GLP, there is a lot of other information available to assist researchers in how to work safely. Much of this will be detailed in the local arrangements for the facility (including standard operating procedures, existing risk assessments, laboratory scripts), safety data sheets (SDSs) for chemicals, instructions for the use of kits in microbiology, user manuals for equipment, etc., and other texts [5,6].

1.3 Risk Assessment

Risk assessment is a tool used to develop ways of working to minimise the risk of causing harm to people and damaging facilities. Carrying out a risk assessment is a fundamental requirement in most health and safety regulations [7–12]. However, this requirement can result in a number of separate assessments being carried out for different parts of the same process, when actually all the requirements could be captured in a single ‘holistic’ risk assessment. Risk assessments must be carried out by ‘competent’ people. (Competent people are those who have sufficient knowledge, ability, training and experience in their field to be able to advise on the safest way to carry out the task that is being assessed.) Principal investigators, laboratory supervisors as well as safety advisors and officers should be able to assist with the process.

• It is pertinent here to differentiate between hazard and risk.
• A hazard is something that has the potential to cause harm.
• A risk is the probability or likelihood of a hazard causing harm.

Before starting work, it is necessary for the people involved to be able to:

• Recognise and identify any hazards associated with the work – these hazards can be associated with materials, equipment, the environment in which it is being done and the people carrying it out – see Table 1.1 for examples of common hazards in laboratories.
• Assess the risks to people posed by the hazards. This includes identifying who could be harmed, how they may be harmed and how severe the harm could be. The hazards that could cause the most severe harm and those that could cause harm to the highest number of people are the ones that must be prioritised when thinking about ways to prevent the harm occurring.
• Reduce and mitigate the risks by adopting ways of working that prevent the hazards coming into contact with people. There is a standard hierarchy of ways to reduce and control hazards, which is shown in Figure 1.1. The most effective way of controlling a hazard is to eliminate it altogether, which is often quite difficult, but must be considered first.
  1. Substitution could involve replacing a substance in one fo...

Table of contents

• Cover
• Table of Contents
• List of Contributors
• Preface
• 1 Principles of Health and Safety and Good Laboratory Practice
• 2 Applications of Chemoinformatics in Drug Discovery
• 3 Bioinformatics and Its Applications in Genomics
• 4 Gene Cloning for the Analysis of Gene Expression
• 5 Proteomic Techniques and Their Applications
• 6 Overproduction, Separation and Purification of Affinity‐Tagged Proteins from Escherichia coli
• 7 Chromatography: Separation Techniques in Biology
• 8 Synthetic Methodology in Chemical Biology
• 9 Reaction Chemical Kinetics in Biology
• 10 Mass Spectrometry and Its Applications
• 11 Applications and Complementarity of Analytical Ultracentrifugation and Light‐Scattering Techniques
• 12 Application of Isothermal Titration Calorimetry (ITC) to Biomolecular Interactions
• 13 An Introduction to Infra‐red and Raman Spectroscopies for Pharmaceutical and Biomedical Studies
• 14 Fluorescence Spectroscopy and Its Applications in Analysing Biomolecular Processes
• 15 Circular Dichroism and Related Spectroscopic Techniques
• 16 Principles and Practice in Macromolecular X‐Ray Crystallography
• 17 Biomolecular NMR Spectroscopy and Structure Determination of DNA
• 18 Cryo‐TEM and Biological Structure Determination
• 19 Computer Modelling and Molecular Dynamics Simulation of Biomolecules
• Index
• End User License Agreement