This presentation provides a knowledge about Toxicology, its types , definition, regulatory guidelines for conducting toxicological studies, OECD guidelines for GLP. This is an assignment in the subject, Pharmacological & Toxicological Screening Methods - II, 2nd Semester, M.Pharm (Pharmacology)
1. Pharmacological & Toxicological Screening Methods - II
“Toxicology |Definition| Types | Regulatory
Guidelines | OECD principles for GLP ”
By
Chetan A., M.Pharm 1st Year (Pharmacology)
K.K. College of Pharmacy
Chennai, TamilNadu
2. Learning Objective
• Introduction
• Types of Toxicology
• Regulatory Guidelines for conducting Toxicity Studies
• OECD Principles of Good Laboratory Practice
• Recent Research
• Facts
• Reference
3. Toxicology
• Toxicology is the branch of science concerned with the nature, effects, detection,
diagnosing and treating of toxins or toxicants.
• The relationship between dose and its effects on the exposed organism is of high
significance in toxicology.
• Toxicology screening is very important for the development of new drugs and for the
extension of the therapeutic potential of existing molecules.
• It leads to more selective in drug therapies to treat disserent diseases such as cancer
having reduced toxic potential to human body.
• It helps to provide protection to humans and environment from toxic effects of
toxicants.
4. History
• Phillip von Hohenheim, also referred to as Paracelcius is known as the “Father of
Toxicology”.
• He is credited with maxim “All things are poisonous and nothing is without poison;
only the dose makes a thing not poisonous.”
• In short “The dose makes the poison”.
• Mathieu Orfila is considered the modern father of toxicology, having given the
subject its first formal treatment in 1813 in his Traité des poisons, also called
Toxicologie générale.
5. Common Terms
• Toxins : A toxin is a poisonous substance produced within living cells or organisms;
synthetic toxicants created by artificial processes are thus excluded.
• Toxicants: A toxicant is any toxic substance. Toxicants can be poisonous and they
may be man-made or naturally occurring. The different types of toxicants can be
found in the air, soil, water, or food.
• Dose: a quantity of a medicine or drug taken or recommended to be taken at a
particular time.
• Adverse effects: An adverse effect is an undesired harmful effect resulting from a
medication or other intervention.
• Toxicity: Toxicity is the degree to which a chemical substance or a particular mixture
of substances can damage an organism.
6. Basic Principles
• The goal of toxicity assessment is to identify adverse effects of a substance. Adverse
effects depend on two main factors:
1.Routes of exposure (oral, inhalation, or dermal) and
2.Dose (duration and concentration of exposure).
• To explore dose, substances are tested in both acute and chronic models.
7. Types of toxicity
1. Acute toxicity - Exposure for a duration of less than 24 hrs, often a single
exposure
2. Subacute toxicity - Repeated exposure for a month or less
3. Subchronic toxicity - Repeated exposure for 1-3 months
4. Chronic toxicity - Repeated exposure for more than 3 months. Usually daily dietry
exposure.
5. Carcinogenecity - Abnormal cell growth and differentiation that can lead to cancer.
The two stages of carcinogenicity are:
• Initiation — a normal cell undergoes irreversible changes.
• Promotion — initiated cells are stimulated to progress to cancer.
8. Continuation..,
6. Developmental toxicity - Related to adverse toxic effects to the developing embryo or
fetus. It can result from toxicant exposure to either parent before conception or to the
mother and her developing embryo or fetus. The three basic types of developmental toxicity
are:
• Embryolethality — failure to conceive, spontaneous abortion, or stillbirth.
• Embryotoxicity — growth retardation or delayed growth of specific organ systems.
• Teratogenicity — irreversible conditions that leave permanent birth defects in live
offspring, such as cleft palette or missing limbs.
7. Genetic toxicity - It results from damage to DNA and altered genetic expression. This
process is known as mutagenesis. The genetic change is referred to as a mutation and the
agent causing the change is called a mutagen. There are three types of genetic changes:
• Gene mutation — change in DNA sequence within a gene.
• Chromosome aberration — changes in the chromosome structure.
• Aneuploidy or polyploidy — increase or decrease in number of chromosomes.
9. Factors that influence chemical toxicity
1. Dosage
• Both large single exposures (acute) and continuous small exposures (chronic) are
studied.
2. Route of exposure
• Ingestion, inhalation or skin absorption
3. Other factors
• Species
• Age
• Sex
• Health
• Environment
• Individual characteristics
10. Routes of Exposure
• There are four routes by which a substance can enter the body:
1.Inhalation,
2.Skin (or eye) absorption,
3.Ingestion, and
4.Injection.
11. Continuation..,
1. Inhalation:
• For most chemicals in the form of vapors, gases, mists, or particulates, inhalation is
the major route of entry. Once inhaled, chemicals are either exhaled or deposited in
the respiratory tract. If deposited, damage can occur through direct contact with
tissue or the chemical may diffuse into the blood through the lung-blood interface.
2. Skin (or eye) absorption:
• Skin (dermal) contact can cause effects such as redness or mild dermatitis; more
severe effects include destruction of skin tissue or other debilitating conditions. Many
chemicals can also cross the skin barrier and be absorbed into the blood system. Once
absorbed, they may produce systemic damage to internal organs.
• Eg, Aniline, Hydrogen cyanide, Organophosphates etc
12. Continuation..,
3. Ingestion:
• Chemicals that accidentally get into the mouth and are swallowed do not generally
harm the gastrointestinal tract itself unless they are irritating or corrosive. Chemicals
that are insoluble in the fluids of the gastrointestinal tract (stomach, small, and large
intestines) are generally excreted. Others that are soluble are absorbed through the
lining of the gastrointestinal tract. They are then transported by the blood to internal
organs where they can cause damage. Eg, Lead
4. Injection:
• Substances may enter the body if the skin is penetrated or punctured by contaminated
objects. Effects can then occur as the substance is circulated in the blood and
deposited in the target organs.
15. Continuation..,
1. Analytical toxicology:
It is the detection, identification, and measurement of foreign compounds
(xenobiotics) in biological and other specimens.
2. Applied toxicology:
It is the early detection of toxicants in the field setting or practice area by the
application of new and modern methods.
3. Clinical toxicology:
The study of diagnosis and treatment of poisoning that can occur in humans.
4. Veterinary toxicology:
The study of diagnosing and treatment of animal poisoning including the
transmission of toxin from animals to humans via animal products (milk, meat & other
food items).
16. Continuation..,
4. Environmental toxicology:
The study of presence of different toxicants including their metabolites and
degradation products in the environment and their effects on humans and animals.
5. Industrial toxicology:
The study of the harmful effects of chemicals used in the workplace, the
products produced by companies, and the wastes created in manufacturing on humans.
6. Reproductive & Development toxicology:
The study of effects of chemicals or toxins on the reproductive system and the
developing embryo.
7. Immuno-toxicology:
The study of effects of toxicants on immune system.
18. Toxicology Branches
• The major areas of specialization are as follows,
I. Mechanistic Toxicology (Basic Biology & Chemistry)
II. Descriptive Toxicology (Testing)
III.Regulatory Toxicology (Rule making & Compliance)
19. I. Mechanistic Toxicology
• It is the branch of toxicology that focuses on how the cellular, biochemical &
molecular mechanisms of chemicals exert toxic effects on living organisms and how
the biological system protects themselved against these adverse effects.
• It aims to identify molecular events that lead from initial exposure to the chemical to
the ultimate manisfestation of toxic injury in an organism.
• Knowledge of the mechanism of toxicity of a substance helps to
1.Enhances the ability to prevent toxicity and design more desirable chemicals.
2.Help governmental regulator to establish legally binding safe limits for human
exposure
3.Forming the basis for therapy and the design of new drugs for treatment of
human disease.
21. Mechanism of Toxicity
1. Delivery of toxicant
2a. Interaction with the target molecule
2b. Alteratiion of biological environment
3. Cellular Dysfunction & Resultant
Toxicity
4. Repair/ Dysrepair/ Adaptation
22. Chemicals that cause Cellular Dysfunction
1. Chemicals that cause DNA adducts - can lead to DNA mutation which can
activate Cell death pathway, if it activates oncogenes or inactivates tumor
suppressors, it can lead to uncontrolled proliferation and cancer. Eg, Benzopyrene.
2. Chemicals that cause protein adducts - can lead to protein dysfunction which can
activate cell death pathway, it can lead to autoimmunity, it can also cause
uncontrolled cell proliferation & Cancer. Eg, Diclofenac glucuronidation
metabolite.
3. Chemicals that cause oxidation stress - can oxidise DNA or protein leading to
DNA mutations or protein dysfunction. Eg, Benzene, CCl4.
4. Chemicals that specifically interact with protein targets - Chemicals that
activates or inactivates ion channels (Na+, Ca2+, K+) needed for neurotransmission
can lead to cellular dysfunction (Tetrodotoxin), chemicals that inhibit cellular
respiration, inhibit proteins /enzymes involved in O2 consumption (Cyanide)
23. Primary metabolic disorders which affect cell survival
1. ATP depletion :
ATP drives ion transporters such as Na+/K+ ATPase, Ca2+ ATPase, biosynthetic reactions, Signal
tansduction regulation, muscle contraction, Cell division, Polymerization of cytoskeleton.
ATP depletion causes
24. Continuation.,
2. Sustained rise in intracellular Ca2+ :
Calcium is involved in Signal transduction regulation, muscle contraction,
Cytoskeletal polymerisation, neurotransmission, enzyme induction, acts in
Ca2+/ATPase, Na/Ca2+ exchanger
• Consequences
1. Depletion of energy reserves
2. Dysfunction of microfilaments
3. Activation of hydrolytic enzymes
4. Generation of ROS/RNS - disintegration of membranes, proteins, DNA etc
25. Continuation.,
3. Overproduction of Reactive Oxygen Species (ROS), Reactive
Nitrogen Species (RNS) :
• ROS can directly oxidize and affect protein functions & can mutate DNA leading to
cellular dysfunction.
• ROS can disrupt mitochondrial membranes & dissipate the electrochemical gradient
needed for ATP synthase.
• NO inhibits cytochrome oxidase.
• ONOO can irreversibly inactivate complexes I/II/III, aconitase, can induce DNA
single strand break.
• ROS/RNS oxidatively inactivates Ca2+/ATPase and elevate Ca2+ & drain ATP
reserves
• Lipid peroxidation, cell swelling & cell rupture
26. II. Descriptive Toxicology
• It is concerned directly with toxicity testing, usuallt on animals and then correlated to
humans, which provides information for safety evaluation and regulatory
requirements.
• It provides dose-response information upon exposure to a harmful toxic agent.
• The results from the toxicity testing are typically applied to approval of product use
and regulating allowable concentrations in the environment.
• The toxicity assessment commonly involves following steps
1.Hazard identification
2.Dose-response assessment
3.Exposure assessment
4.Risk characterization
27. Continuation.,
1. Hazard identification :
• It determines the exposure to chemical can increase the incidents of a particular
adverse effect.
• It is done by,
Hazard identification data
Human epidemiology data
Animal bioassay
Supporting data
Prediction of hazard-structure activity relationship
28. Continuation.,
2. Dose-Response Assessment
• Relationship between the dose and the corresponding response produced.
• There are two types of dose response relationship
a. Graded dose response relationship : It describes the response of an individual
organism to varying dose of chemicals
b. Quantal dose response relationship : characterizes of the distribution of intensity
of the effect to different doses in a population of individual organisms.
31. Continuation.,
3. Exposure Assessment :
• Exposed population (General public or selected groups).
• Types of substances (pharmaceutical, chemicals or environmental pollutants)
• Single substance or mixture of substances
• Duration of exposure
• Pathway and media
32. Continuation.,
4. Risk Characterization :
• Review toxicity and exposure assessment output
• Quantity risks
• Combine risks across all pathways
• Assess and preaent uncertainities
• Consider site specific human studies
33. Toxicity Testing Methods
1. Acute toxicity testing : Single dose in one or two different animal species for 14
days
2. Subacute toxicity testing : Repeated dose for upto 90 days.
3. Chronic toxicity testing : Repeated dose with minimum of one rodent & one
nonrodent for more than 90 days
34. Importance of Toxicity Testing
• To have an idea of toxic doses of xenobiotics for certain organisms.
• Evolution of safe doses of those toxicants for certain organisms
• Recommendation of maximum permissible limits of those substances in the ambient
air and drinking water
• The data on long term toxicity tests may be reliable for the evaluation of safe levels
of toxicants
• Evolution and recommendation of maximum acceptable daily intake
• Developing the air water quality
35. III. Regulatory Toxicology
• It is the branch of toxicology that deals with the relationship between the discipline
of toxicology and regulatoryinstitutions.
• The health of humans relies on toxicological principal and toxicity evaluation data to
formulate a decision.
• Main Focus
• The authority has to take a decision on Acceptable Daily Intake (ADI) of a chemical
so that quantity of that chemical exposure is adjusted safe in terms of health.
• The authority also have the power to formulate some law or regulatory roles and to
implement them rigidly.
36. Continuation.,
• Principle of toxicology says “no chemical is safe” all chemicals are potentially toxic
depending upon their exposure, concentration , time, frequency and nature.
• Regulators formulate the threshold doses to reduce exposure concentration so that
risks can be minimized to highest level.
• The regulatory authorities are WHO, ICH, EPA, OECD, FDA
37. 1. ICH - International Conference on Harmonisation
• ICH & WHO has produced a comprehensive set of safety Guidelines to uncover potential risks
like carcinogenicity, genotoxicity and reprotoxicity.A recent breakthrough has been a non-
clinical testing strategy for assessing the QT interval prolongation liability(most important reason
now a days to withdraw drug from market).
1. CARCINOGENICITYSTUDIES
a. S1A:NeedforCarcinogenicityStudiesofPharmaceuticals
• Thisdocument provides a consistent definition of the circumstances under which it isnecessary to
undertake carcinogenicity studies on new drugs. These recommendations take into account the known
riskfactors as well as the intended indicationsand durationof exposure.
b. S1B:TestingforCarcinogenicityofPharmaceuticals
• This documentprovides guidance on the need to carry out carcinogenicity studies in both mice and rats.
c. S1C(R2):DoseSelectionforCarcinogenicityStudiesof Pharmaceuticals
• Thisdocument addresses the criteria for the selection of the high dose to be used in carcinogenicity
studies on new therapeutic agents to harmonize current practices and improve the design of studies.
38. Continuation.,
2. GENOTOXICITYSTUDIES
• S2(R1):GuidanceonGenotoxicityTestingandDataInterpretationfor PharmaceuticalsIntended forHumanUseS2A,
S2B
a. S2A:GuidanceonSpecificAspectsofRegulatoryGenotoxicityTestsfor Pharmaceuticals
• Thisdocument provided specific guidance and recommendations for in vitroand in vivotestsand on
the evaluationof testresults.
b. S2B:Genotoxicity:AStandardBatteryforGenotoxicityTestingforPharmaceuticals
• Thisdocument addressed two fundamental areas of genotoxicity testing: the identification of a standard
set of assays to be conducted for registration, and the extent of confirmatory experimentation in any
particular genotoxicity assay in the standardbattery.
39. Continuation.,
3. TOXICOKINETICSANDPHARMACOKINETICS
a. S3A:NoteforGuidanceonToxicokinetics:TheAssessmentofSystemic ExposureinToxicityStudies
• Thisdocument gives guidance on developing test strategies in toxicokinetics and the need to integrate
pharmacokinetics into toxicitytesting.
b. S3B:Pharmacokinetics:GuidanceforRepeatedDoseTissueDistribution Studies
• Thisdocument gives guidance on circumstances when repeated dose tissuedistributionstudies shouldbe
considered (i.e.,when appropriate data cannot be derived from othersources).
4. TOXICITYTESTING
a. S4:DurationofChronicToxicityTestinginAnimals(RodentandNonRodentToxicityTesting)
• The text incorporates the guidance for repeat-dose toxicity teststhat was agreed at the time of ICH 1, in
1991(reduction of the duration ofrepeat dose toxicity studies in the rat from 12 to6 months).
40. Continuation.,
5. REPRODUCTIVETOXICOLOGY
• S5(R2):DetectionofToxicitytoReproductionforMedicinal Products&ToxicitytoMale FertilityS5A,S5B(M)
• This document provides guidance on testsfor reproductive toxicity.Itdefines the periods of treatmentto
be usedinanimalsto better reflect human exposure to medical products and allow morespecific
identificationof stagesat risk.
6. BIOTECHNOLOGICALPRODUCTS
• S6(R1):PreclinicalSafetyEvaluationofBiotechnology-Derived Pharmaceuticals
• Thisdocument coversthe pre-clinical safetytestingrequirementsfor biotechnological products.
7. PHARMACOLOGYSTUDIES
• S7A:SafetyPharmacologyStudiesforHumanPharmaceuticals
• Thisdocument addressesthe definition,objectives and scope of safety pharmacology studies.
• S7B:TheNon-ClinicalEvaluationofthePotentialforDelayedVentricular Repolarization(QTIntervalProlongation)
byHuman
• This Guidelinedescribes a non-clinical testing strategy for assessingthe potentialof a test substance to
delayventricular repolarization.
41. Continuation.,
8. IMMUNOTOXICOLOGYSTUDIES
• S8:ImmunotoxicityStudiesforHuman
• This Guideline addresses the recommendations on nonclinical testing for immunosuppression induced by
low molecular weight drugs(non-biologicals).
9. NONCLINICALEVALUATIONFORANTICANCER PHARMACEUTICALS
• S9:NonclinicalEvaluationforAnticancer
• ThisGuideline provides information for pharmaceuticals that are only intended to treat cancer in
patients with late stage or advanced disease regardless of the route of administration, including both small
molecule and biotechnology-derivedpharmaceuticals.
10. PHOTOSAFETYEVALUATION
• S10:PhotosafetyEvaluationofPharmaceuticals(DraftDocument)
• TheS10 draft Guideline has been released forconsultation under Step 2 of the ICHprocess in November
2012.
42. 2. OECD - Organisation for Economic Co-operation and Development
• Themissionof the OECD isto promote policies that willimprove the economic and social well-
being of people around the world.
• The OECD provides a forum in which governments can work together to share experiences and
seek solutionsto commonproblems.
• GUIDELINES:
• 402Acute DermalToxicity
• 403Acute InhalationToxicity
• 404Acute DermalIrritation/Corrosion
• 405Acute EyeIrritation/Corrosion
• 406SkinSensitisation
• 407RepeatedDose 28-dayOral ToxicityStudyinRodents
• 408RepeatedDose 90-DayOral ToxicityStudyin Rodents
• 409RepeatedDose 90-DayOral ToxicityStudyin Non-Rodents
• 410Repeated Dose Dermal Toxicity:21/28-dayStudy
46. 3. EPA - Environment Protection Agencies Guidlines
1. Acute toxicity test guidelines (GroupA)
2. Subchronic toxicity test guidelines (Group B)
3. Chronic toxicity test guidelines (Group C)
4. Genetic toxicity test guidelines (Group D)
5. Neurotoxicity test guidelines (Group E)
6. Special study test guidelines (Group F)
7. Health effect chemical specific test guideline (Group G)
8. Supplemental guidance.
54. OECD
• The Organisation for Economic Co-operation and Development (OECD) is an
intergovernmental organisation (OECD Secretariat, located in Paris, France)
• Representatives of 29 industrialised countries in North America, Europe and the
Pacific, as well as the European Commission, meet to co-ordinate and harmonize
policies, discuss issues of mutual concern, and work together to respond to
international problems.
• Most of the OECD’s work is carried out by more than 200 specialised Committees
and subsidiary groups composed of Member country delegates.
• The work of the OECD related to chemical safety is carried out in the Environmental
Health and Safety Division.
55. Good Laboratory Practice
• Good Laboratory Practice (GLP) is a quality system concerned with the
organizational process and the conditions under which non-clinical health and
environmental safety studies are planned, performed, monitored, recorded, archived
and reported.
• Non-clinical health and environmental safety study referred to simply as "study",
means an experiment or set of experiments in which a test item is examined under
laboratory conditions or in the environment to obtain data on its properties and/or its
safety, intended for submission to appropriate regulatory authorities.
• The Principles of Good Laboratory Practice (GLP) have been developed to promote
the quality and validity of test data used for determining the safety of chemicals &
Chemical products
56. History
• In the early 70’s FDA became aware of cases of poor laboratory practice all over the
United States.
• They discovered a lot fraudulent activities and a lot of poor lab practices.
• 1. Equipment not been calibrated to standard form therefore giving wrong
measurements.
• 2. Incorrect/inaccurate accounts of the actual lab study.
• 3. Inadequate test systems .
• One investigation- made headline news
• Lab - IBT
• Mice that they had used to test developed cancer and died.
• IBT lab threw the dead mice and covered results deeming the products good for
human consumption.
57. Continuation
• Government and industry became concerned about the quality of non-clinical health
and environmental safety studies upon which hazard assessments are based. As a
consequence, OECD Member countries have established criteria for the performance
of these studies.
• In 1979 and 1980, an international group of experts established under the Special
Programme on the Control of Chemicals developed the “OECD Principles of Good
Laboratory Practice” (GLP). These Principles of GLP were adopted by the OECD
Council in 1981.
• In 1995 and 1996, a new group of experts was formed to revise and update the
Principles.
• The purpose of these Principles of Good Laboratory Practice is to promote the
development of quality test data. Comparable quality of test data forms the basis for
the mutual acceptance of data among countries.
58. GLP in India
• National GLP-compliance Monitoring Authority was established by the Department
of Science & Technology
• Approval of the Union Cabinet on April 24, 2002
• A provisional member of the OECD for GLP.
• India is an Observer to the OECD’s Working Group on GLP
• The Authority has trained 33 experts in the country as GLP inspectors
59. Scope
• These Principles of Good Laboratory Practice should be applied to the non-clinical
safety testing of test items contained in pharmaceutical products, pesticide products,
cosmetic products, veterinary drugs as well as food additives, feed additives, and
industrial chemicals
• Test items are frequently synthetic chemicals, but may be of natural or biological
origin
• Unless specifically exempted by national legislation, these Principles of Good
Laboratory Practice apply to all non-clinical health and environmental safety studies
for the purpose of registering or licensing pharmaceuticals, pesticides, food and feed
additives, cosmetic products, veterinary drug products and similar products, and for
the regulation of industrial chemicals.
60. Objectives of GLP
• Data submitted are a true reflection of the results that are
• obtained during the study.
• Not to indulge in any fraud activity by labs.
• International acceptance of tests
61. GLP Principles
1. Test Facility Organization and Personnel
2. Quality Assurance Program
3. Facilities
4. Apparatus, Material, and Reagents
5. Test Systems
6. Test and Reference Items
7. SOPs
8. Performance of the Study
9. Reporting of Study Results
10. Storage and Retention of Records and Materials
62. 1. Test Facility Organization and Personnel
A. Test Facility Management’s Responsibilities
B. Study Director’s Responsibilities
C. Principal Investigator’s Responsibilities
D. Study Personnel’s Responsibilities
63. Continuation
A. Test Facility Management’s Responsibilities
• Each test facility management should ensure that these Principles of Good
Laboratory Practice are complied with, in its test facility.
• To ensure that a sufficient number of qualified personnel, appropriate facilities,
equipment, and materials are available for the timely and proper conduct of the
study
• To ensure that appropriate and technically valid Standard Operating Procedures are
established and followed, and approve all original and revised Standard Operating
Procedures;
• To ensure the maintenance of a record of the qualifications, training, experience and
job description for each professional and technical individual
• To ensure that test facility supplies meet requirements appropriate to their use in a
study.
64. Continuation
B. Study Director’s Responsibilities
• The Study Director has the responsibility for the overall conduct of the study and for
its final report.
• To approve the study plan and any amendments to the study plan by dated signature
• To ensure that all raw data generated are fully documented and recorded
• To ensure that the procedures specified in the study plan are followed, and assess and
document the impact of any deviations from the study plan on the quality and
integrity of the study, and take appropriate corrective action if necessary;
acknowledge deviations from Standard Operating Procedures during the conduct of
the study
• To ensure that computerised systems used in the study have been validated
65. Continuation
C. Principal Investigator’s Responsibilities
• The Principal Investigator will ensure that the delegated phases of the study are
conducted in accordance with the applicable Principles of Good Laboratory Practice.
D. Study Personnel’s Responsibilities
• All personnel involved in the conduct of the study must be knowledgeable in those
parts of the Principles of Good Laboratory Practice which are applicable to their
involvement in the study.
• Study personnel should exercise health precautions to minimize risk to themselves
and to ensure the integrity of the study. They should communicate to the appropriate
person any relevant known health or medical condition in order that they can be
excluded from operations that may affect the study.
66. Continuation
• All study personnel are responsible for recording raw data promptly and accurately
and in compliance with these Principles of Good Laboratory Practice, and are
responsible for the quality of their data
• Study personnel will have access to the study plan and appropriate Standard
Operating Procedures applicable to their involvement in the study. It is their
responsibility to comply with the instructions given in these documents
67. 2. Quality Assurance Program
A. General
• The Quality Assurance Programme should be carried out by an individual or by
individuals designated by and directly responsible to management and who are
familiar with the test procedures.
B. Responsibilities of the Quality Assurance Personnel
• To maintain copies of all approved study plans and Standard Operating Procedures
in use in the test facility
• To verify that the study plan contains the information required for compliance with
these Principles of Good Laboratory Practice. This verification should be
documented
• To conduct inspections to determine that study plans and Standard Operating
Procedures have been made available to study personnel and are being followed.
68. 3. Facilities
A. General
• The test facility should be of suitable size, construction and location to meet the
requirements of the study and to minimize disturbance that would interfere with the
validity of the study.
B. Test System Facilities
• There should be storage rooms or areas as needed for supplies and equipment.
Storage rooms or areas should be separated from rooms or areas housing the test
systems and should provide adequate protection against infestation, contamination,
and/or deterioration
• The test facility should have a sufficient number of rooms or areas to assure the
isolation of test systems and the isolation of individual projects, involving substances
or organisms known to be or suspected of being biohazardous.
69. Continuation
C. Facilities for Handling Test and Reference Items
• To prevent contamination or mix-ups, there should be separate rooms or areas for
receipt and storage of the test and reference items, and mixing of the test items with a
vehicle
D. Archive Facilities
• Archive facilities should be provided for the secure storage and retrieval of study
plans, raw data, final reports, samples of test items and specimens. Archive design
and archive conditions should protect contents from un-timely deterioration.
E. Waste Disposal
• Handling and disposal of wastes should be carried out in such a way as not to
jeopardise the integrity of studies. This includes provision for appropriate collection,
storage and disposal facilities, and decontamination and transportation procedures.
70. 4. Apparatus, Material, and Reagents
• Apparatus, including validated computerized systems, used for the generation,
storage and retrieval of data, and for controlling environmental factors relevant to the
study should be suitably located and of appropriate design and adequate capacity.
• Apparatus and materials used in a study should not interfere adversely with the test
systems.
• Chemicals, reagents, and solutions should be labelled to indicate identity (with
concentration if appropriate), expiry date and specific storage instructions.
Information concerning source, preparation date and stability should be available.
The expiry date may be extended on the basis of documented evaluation or analysis.
71. 5. Test System
A. Physical/Chemical
• Apparatus used for the generation of physical/chemical data should be suitably
located and of appropriate design and adequate capacity.
B. Biological
• Proper conditions should be established and maintained for the storage, housing,
handling and care of biological test systems, in order to ensure the quality of the data.
• Newly received animal and plant test systems should be isolated until their health
status has been evaluated.
• All information needed to properly identify the test systems should appear on their
housing or containers. Individual test systems that are to be removed from their
housing or containers during the conduct of the study should bear appropriate
identification, wherever possible.
72. 6. Test and Reference Items
A. Receipt, Handling, Sampling and Storage
• Records including test item and reference item characterization, date of receipt,
expiry date, quantities received and used in studies should be maintained.
• Storage container(s) should carry identification information, expiry date, and specific
storage instruction
B. Characterisation
• Each test and reference item should be appropriately identified (e.g., code, name,
biological parameters).
• The stability of test and reference items under storage and test conditions should be
known for all studies.
73. 7. Standard Operating Procedures
• A test facility should have written Standard Operating Procedures approved by test
facility management that are intended to ensure the quality and integrity of the data
generated by thatb test facility.
• Deviations from Standard Operating Procedures related to the study should be
documented and should be acknowledged by the Study Director and the Principal
Investigator(s), as applicable
74. 8. Performance of the Study
A. Study Plan
• For each study, a written plan should exist prior to the initiation of the study. The
study plan should be approved by dated signature of the Study Director and verified
for GLP compliance by Quality Assurance personnel as specified in Section 2.2.1.b
• For short-term studies, a general study plan accompanied by a study specific
supplement may be used.
• Amendments to the study plan should be justified and approved by dated signature of
the Study Director and maintained with the study plan.
B. Content of the Study Plan
• Identification of the Study, the Test Item and Reference Item
• Information Concerning the Sponsor and the Test Facility
• Dates, Test methods
• Issues and records
75. Continuation
C. Conduct of the Study
• A unique identification should be given to each study.
• All data generated during the conduct of the study should be recorded directly,
promptly, accurately, and legibly by the individual entering the data.
• Data generated as a direct computer input should be identified at the time of data
input by the individual(s) responsible for direct data entries
76. 9. Reporting of Study Results
A. General
• A final report should be prepared for each study.
• Reports of Principal Investigators or scientists involved in the study should be signed
and dated by them.
• Corrections and additions to a final report should be in the form of amendments.
B. Content of the Final Report
• Identification of the Study, the Test Item and Reference Item.
• Information Concerning the Sponsor and the Test Facility.
• Description of Materials and Test Methods
77. 10. Storage and Retention of Records and Materials
A. The following should be retained in the archives for the period specified by the
appropriate authorities:
• The study plan, raw data, samples of test and reference items, specimens, and the
final report of each study;
• Records of all inspections performed by the Quality Assurance Programme, as well
as master schedules;
• Records of qualifications, training, experience and job descriptions of personnel;
• Records and reports of the maintenance and calibration of apparatus
• The historical file of all Standard Operating Procedures
B. Material retained in the archives should be indexed so as to facilitate orderly storage
and retrieval.
78. Benefits of GLP
• It will give better image of company as a Quality producer in Global market.
• Provide hot tips on analysis of data as well as measure uncertainty and perfect record
keeping.
• Provide guideline for doing testing and measurement in detail
• Provide guidelines and better control for maintenance of instruments, environment
control, preservation of test records etc
• Conclusion
• GLP is an FDA regulation which is accepted and approved as international standards
by OECD to avoid the fraud activities of the testing laboratories for pesticides ,
pharmaceuticals , food additives , dyes, to save the human and environmental health
and also erect good international trade and establish good relationship among the
countries
79. Recent Research
1. Inclusion of Positive Self-reporting by Mothers of Substance Exposed Neonates Increases the Predictability
of NAS Severity Over Toxicology Alone - Danielle Roth et,al. (2020)
• Objective: The rise in opioid use among pregnant women has resulted in an increase in the incidence of neonatal
abstinence syndrome (NAS). Despite the focus on opioid use, prenatal polysubstance exposure is often associated
with NAS diagnosis and severity. Drug toxicology screens such as urine drug screens and umbilical cord
toxicology are dependent upon the substance, timing, frequency, and dose to detect substances present and can
underestimate the neonatal exposure. The aim of this study was to identify the predictability of the consequences
of prenatal polysubstance exposure versus opioid only exposure based on toxicology and toxicology plus self-
report.
• Methods: Neonates > 35 weeks gestation with prenatal opioid exposure were included in this retrospective data
analysis. NAS was identified using maternal urine drug screen (UDS) toxicology, self-reported exposure during
pregnancy, and neonatal toxicology. Analysis was conducted using Stata 15.1 utilizing McNemar’s test, chi-
square for categorical outcomes, and Wilcoxon test for numerical outcomes.
• Results: A statistically significant difference in length of stay and length of treatment with poly-exposed neonates
was observed when maternal self-report was considered with toxicology, but not with toxicology alone. This
trend was observed for cumulative hospital length of stay as well as length and dose of treatment.
80. Facts
• Raw Bitter almonds contain traces of cyanide – the lethal dose for an adult is about
50 almonds, whereas a child would only need to consume 5-10 almonds.
• LD50 is a test that finds an average dose required to kill half of the animals being
tested, normally rats in human toxicity.
• The Fixed Dose Procedure (LD50 alternative) measures oral toxicity in a similar way,
but using fewer animals and with less suffering.
• Minimum lethal dose (MLD, also LDmin), the smallest amount of drug that can
produce death in an animal species under controlled conditions.
• Over 90% of the poisonings are accidental, and could have been prevented.
• Vegetables like Tomato, Peppers, Eggplant & Cauliflower contain Nicotine (as
defensive poison) in small amounts, which is linked with decreased risk of
developing Parkinson’s disease.
