Good laboratory practises

GOOD LABORATORY
PRACTISES
PRATHYUSHA
Msc BIOTECHNOLOGY
SCHOOL OF BIOSCIENCES
MAHATMA GANDHI UNIERSITY

INTRODUCTION
 Good Laboratory Practice (GLP) regulations.
 Became part of the regulatory landscape in
the latter part of the 1970s.
 In response to malpractice in research and
development (R&D)activities by
pharmaceutical companies and contract
facilities used by them.
 The malpractice included cases of fraud,
lack of proper management and organization
of studies performed to generate data for

 The US Food and Drug Administration (FDA)
mounted a series of investigations in
toxicology laboratories throughout the USA.
 The results of these investigations revealed
a situation that could only be dealt with by
imposing binding regulations.
 These regulations are the GLP regulations.
GLP regulations were first instituted by US
FDA, then by US Environmental Protection
Agency (EPA); many other nations have
since followed suit.

 In 1981, the Organization for Economic
Cooperation and Development (OECD)
also published GLP Principles, and these
now dominate the international arena.
 To date 30 countries (the member states
of the OECD) have signed an agreement
binding them to OECD GLP Principles.
 Other non-OECD member states have
also adopted the OECD GLP Principles.

THE FUNDAMENTAL POINTS OF
GLP
 The GLP regulations set out the rules for
good practice and help researchers perform
their work in compliance with their own pre-
established plans and standardized
procedures.
 The regulations are not concerned with the
scientific or technical content of the research
programmes.
 Nor do they aim to evaluate the scientific
value of the studies.

 All GLP texts, irrespective of their origin,
stress the importance on the following points
five points:
 1. Resources: organization, personnel,
facilities and equipment
 2. Characterization: test items and test
systems
 3. Rules: study plans (or protocols) and
written procedures
 4. Results: raw data, final report and
archives
 5. Quality Assurance.

1. Resources
 Organization and personnel
◦ GLP regulations require that the structure of R&D
organizations and the responsibilities of R&D
personnel be clearly defined.
◦ GLP also stresses that there should be sufficient
staff to perform the tasks required.
◦ The qualifications and the training of staff must
also be defined and documented.
 Facilities and equipment
◦ The regulations emphasize the need for sufficient
facilities and equipment to perform the studies.
◦ All equipment must be in working order. To
ensure this ,a strict programme of qualification,
calibration and maintenance must be adopted.

2. Characterization
 In order to perform a study correctly, it is
essential to know as much as possible about
the materials used during the study.
 For studies that evaluate the properties of
pharmaceutical compounds during non-
clinical studies, it is a prerequisite to have
details about the test item and the test
system (often an animal or plant) to which
the test item is to be administered.

3. Rules
Protocols and written procedures
The main steps of research studies are prescribed in
the study plan or protocol.
 Being able to repeat studies and obtain similar
results is a sine qua non of mutual acceptance of
data and, indeed, a central tenet of the scientific
method, so the details of routine procedures must
also be available to scientists involved in the study.
 However, the protocol, which provides the
experimental design and timeframe for the study,
does not contain all the technical detail necessary to
conduct the study. These details are found in written
standard operating procedures (SOPs).
 With the protocol and the SOPs it should be possible
to repeat the study exactly, if necessary.

4. Results
Raw data
 All studies generate raw data. These are the outcome
of research and form the basis for establishing
scientific interpretations and arriving at conclusions.
 The raw data must also reflect the procedures and
conditions of the study.
Final Report
 The study report contains an account of the way in
which the study was performed, incorporates the
study results and includes the scientific interpretation
of the data.
 The report is provided to regulatory authorities as part
of the submission for registration and marketing
approval.
Archives

5. Quality Assurance
 Quality assurance (QA), as defined by
GLP, is a team of persons (often called
the Quality assurance unit – QAU)
charged with assuring management that
GLP compliance has been attained within
the laboratory.
 QA must be independent from scientists
involved in the operational aspects of the
study being performed.
 QA functions as a witness to the whole
non-clinical research process.

Chemicals and reagents
 Chemicals, reagents and reference
substances play a vital role in the
correctness of laboratory results.
 Microbiological cultures and media.
 Traceability in all records.
 Uncontrolled cost cutting in this area
could lead to increased number of
laboratory deviations.

Media Preparation and Quality
Control
 The quality of work in a microbiological
laboratory depends on the quality of the
culture media.
 It is essential to use the correct media for the
purpose at hand, although the correct media
is not always obvious.
◦ For example, water testing is commonly done
with R2A agar, but many facilities use TSA for this
purpose.
 The recommendation is provided that the
choice of media should be consistent,

 The recommendations include accurate weighing
of dehydrated components, the use of high quality
(USP Purified) water, completely dissolving the
dehydrated media or individual ingredients, and
the need to control the heating of the media to
avoid damaging heat-labile components of the
media.
 Some recommendations on the labelling and
packaging of media are also provided.
 Excesses of heat and cold are to be guarded
against, as is the potential for dehydration of
poured plates.
 Some guidance is also provided in quality control
for molten media used in pour plates.

Maintenance of Microbiological
Cultures
 Second only to media, safeguarding the
stock cultures is the most important
component of a successful microbiology
laboratory.
 These must be handled carefully at all times
to avoid contamination.
 The care of the cultures starts upon receipt.
 A careful stock culture curator will confirm
the identity of the received cultures, even if
they come from as respected a source as a
national culture collection.
 Mistakes can happen.
 The use of an incorrect strain in a
compendia test could bring the results of

AGAR SLANT
SALINE
SUSPENSI
ON
LIQUID NITROGEN

Microbiology Lab Practices and
Safety Rules

EQUIPMENTS IN MICROBIOLOGY LABORATORY
ANALYTICAL BALANCER REFRIGERATOR
SHAKING INCUBATORS

DEEP FREEZER
INVERTED PHASE CONTRAST
MICROSCOPY

Magnetic stirrer is a device which provides mixing and keeping the
chemical solutions and mixtures at a certain time and temperature by the
help of a magnetic bar.
Vortex agitates the solutions in the tube, flask and so on in certain speed and
duration.
MAGNETIC STIRRER & VORTEX

Laboratory Safety Equipment
Biological Safety Cabinet
 A biological safety cabinet (BSC) is used as a primary
barrier against exposure to infectious biological
agents.
 A BSC has High Efficiency Particulate Air (HEPA)
filters. The airflow in a BSC is laminar, i.e. the air
moves with uniform velocity in one direction along
parallel flow lines.
 BSCs are not chemical fume hoods. A percentage of
the air is recirculated in most types of BSCs.
 HEPA filters only trap particulates, allowing any
contaminant in non-particulate form to pass through
the filter.

Proper use of BSCs
1. Operate the cabinet for five minutes before and after
performing any work in it in order to purge airborne
contaminants.
2. Before and after use, wipe the surface of the BSC
with a suitable disinfectant, e.g., 70% alcohol or a
10% bleach solution.
3. Place everything you will need inside the cabinet
before beginning work, including a waste container.
You should not have to penetrate the air barrier of the
cabinet once work has begun.
4. Do not place anything on the air intake grills, as this
will block the air supply.
5. You should prevent unnecessary opening and closing
of door because this will disrupt the airflow of the

6. Always wear a lab coat while using the cabinet
and conduct your work at least four inches inside
the cabinet.
7. Place burners to the rear of the cabinet to reduce
air turbulence.
8. Do not work in the BSC while the ultraviolet light
is on. Ultraviolet light can quickly injure the eye.
9. When finished with your work procedure,
decontaminate the surfaces of any equipment.
10. Remove the equipment from the cabinet and
decontaminate the work surface.
11. Thoroughly wash your hands and arms.

Bio safety Levels and
Practices
 The Centres for Disease Control (CDC)
and the National Institutes of Health (NIH)
have developed standard procedures
providing protection against biological
hazards.
 Bio safety levels are selected to provide
the end-user with a description of the
minimum containment required for
handling different microorganisms safely
in a laboratory setting and reduce or

 Containment refers to safe methods for
managing infectious material in the
laboratory environment.
 These bio safety levels are applicable to
facilities such as diagnostic, research,
clinical, teaching, and production facilities
that are working at a laboratory scale. The
four bio safety levels are described as:

Bio safety Level 1 (BSL1)
 Examples of BSL1 Agents: Bacillus subtilus,
Naegleria gruberi, many Escherichia coli, Infectious
Canine Hepatitis Virus.
 BSL1 containment is suitable for work involving well-
characterized agents not known to cause disease in
healthy adult humans, and of minimal potential
hazard to laboratory personnel and the environment.
 A BSL1 lab requires no special design features
beyond those suitable for a well-designed and
functional laboratory.
 Biological safety cabinets (BSCs) are not required.
 Work may be done on an open bench top, and
containment is achieved through the use of practices
normally employed in a basic microbiology laboratory.

 Examples of BSL2 Agents: Bacillus anthracis,
Bordetella pertussis, Brucella spp., Cryptococcus
neoformans, Clostridium botulinum, Clostridium
tetani, Helicobacter pylori, most Salmonella spp.,
Yersinia pestis, Mycobacterium leprae, Shigella
spp., Human Immunodeficiency Virus, Human
blood.
 The primary exposure hazards associated with
organisms requiring BSL2 are through the
ingestion, inoculation and mucous membrane
route.
 Agents requiring BSL2 facilities are not generally
transmitted by airborne routes, but care must be
taken to avoid the generation of aerosols
(aerosols can settle on bench tops and become

 Primary containment devices such as
BSCs and centrifuges with sealed rotors
or safety cups are to be used as well as
appropriate personal protective
equipment (i.e., gloves, laboratory coats,
protective eyewear).
 As well, environmental contamination
must be minimized by the use of hand
washing sinks and decontamination
facilities (autoclaves).

 Examples of BSL3 Agents: Myobacterium
tuberculosis, Salmonella typhi, Vesicular
Stomatitis Virus, Yellow Fever Virus,
Francisella tularensis, Coxiella burnetti.
 Laboratory personnel have specific training
in handling these pathogenic and
potentially lethal agents and are
supervised by scientists who are
experienced in working with these agents.
 These agents may be transmitted by the
airborne route, often have a low infectious
dose to produce effects and can cause
serious or life-threatening disease.

 BSL3 emphasizes additional primary and
secondary barriers to minimize the
release of infectious organisms into the
immediate laboratory and the
environment.
 Additional features to prevent
transmission of BSL3 organisms are
appropriate respiratory protection, HEPA
filtration of exhausted laboratory air and
strictly controlled laboratory access.

 Examples of BSL5 Agents: smallpox virus, Ebola
virus, hemorrhagic fever viruses.
 This is the maximum containment available and is
suitable for facilities manipulating agents that are
dangerous/exotic agents, which post a risk of
life threatening disease.
 These agents have the potential for aerosol
transmission, often have a low infectious dose
and produce very serious and often fatal
disease; there is generally no treatment or
vaccine available.

 This level of containment represents an
isolated unit, functionally and, when
necessary, structurally independent of other
areas.
 BSL4 emphasizes maximum containment of
the infectious agent by complete sealing of
the facility perimeter with confirmation by
pressure decay testing; isolation of the
researcher from the pathogen by his or her
containment in a positive pressure suit or
containment of the pathogen in a Class III
BSC line; and decontamination of air and

CONCLUSION
 Good Laboratory Practice (GLP)
regulations are applied to non-clinical
safety of study items contained in
pharmaceutical products, cosmetic
products, veterinary drugs, devices as
well as food additives. The purpose of
testing these items is to obtain information
on their safety with respect to human
health and environment. GLP is also
required for registration purpose and
licensing of pharmaceuticals, pesticides,
food additives, veterinary drug products

Good laboratory practises

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