Toxicology | Definition | Types | Regulatory Guidelines | OECD Principles for GLP

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 1 st 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 T he goal of toxicity assessment is to identify adverse effects of a substance. Adverse effects depend on two main factors: R outes of exposure (oral, inhalation, or dermal) and Dose (duration and concentration of exposure). To explore dose, substances are tested in both acute and chronic models.

7. Types of toxicity Acute toxicity - Exposure for a duration of less than 24 hrs, often a single exposure Subacute toxicity - Repeated exposure for a month or less Subchronic toxicity - Repeated exposure for 1-3 months Chronic toxicity - Repeated exposure for more than 3 months. Usually daily dietry exposure. 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.., 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. Genetic toxicity - It r esults 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 Dosage Both large single exposures (acute) and continuous small exposures (chronic) are studied. Route of exposure Ingestion, inhalation or skin absorption 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: I nhalation, S kin (or eye) absorption, I ngestion, and I njection.

11. Continuation.., 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. 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.., 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 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.

13. Types of Toxicology

14. General types of Toxicology Analytical toxicology Applied toxicology Clinical toxicology Forensic toxicology Environment toxicology Industrial toxicology Reproductive & Developmental toxicology Immuno-toxicology

15. Continuation.., Analytical toxicology: It is the detection, identification, and measurement of foreign compounds (xenobiotics) in biological and other specimens. Applied toxicology: It is the early detection of toxicants in the field setting or practice area by the application of new and modern methods. Clinical toxicology: The study of diagnosis and treatment of poisoning that can occur in humans. 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.., 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. 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. Reproductive & Development toxicology: The study of effects of chemicals or toxins on the reproductive system and the developing embryo. Immuno-toxicology: The study of effects of toxicants on immune system.

17. Branches of Toxicology

18. Toxicology Branches The major areas of specialization are as follows, Mechanistic Toxicology (Basic Biology & Chemistry) Descriptive Toxicology (Testing) 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 Enhances the ability to prevent toxicity and design more desirable chemicals. Help governmental regulator to establish legally binding safe limits for human exposure Forming the basis for therapy and the design of new drugs for treatment of human disease.

20. Mechanism of Toxicity Exposure (Acute or repeated Oral, Inhalation, Skin ) Pharmacokinetics (ADME) Toxic effects System/Organ level (CNS, lung, reproductive system etc) Cellular/Biochemical level Molecular level (DNA, RNA, protein)

21. Mechanism of Toxicity Delivery of toxicant 2a. Interaction with the target molecule 2b. Alteratiion of biological environment Cellular Dysfunction & Resultant Toxicity Repair/ Dysrepair/ Adaptation

22. Chemicals that cause Cellular Dysfunction 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. 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. Chemicals that cause oxidation stress - can oxidise DNA or protein leading to DNA mutations or protein dysfunction. Eg, Benzene, CCl 4 . Chemicals that specifically interact with protein targets - Chemicals that activates or inactivates ion channels (Na + , Ca 2+ , K + ) needed for neurotransmission can lead to cellular dysfunction (Tetrodotoxin), chemicals that inhibit cellular respiration, inhibit proteins /enzymes involved in O 2 consumption (Cyanide)

23. Primary metabolic disorders which affect cell survival ATP depletion : ATP drives ion transporters such as Na + /K + ATPase, Ca 2+ ATPase, biosynthetic reactions, Signal tansduction regulation, muscle contraction, Cell division, Polymerization of cytoskeleton. ATP depletion causes

24. Continuation ., Sustained rise in intracellular Ca 2+ : Calcium is involved in Signal transduction regulation, muscle contraction, Cytoskeletal polymerisation, neurotransmission, enzyme induction, acts in Ca 2+ /ATPase, Na/Ca 2+ exchanger Consequences Depletion of energy reserves Dysfunction of microfilaments Activation of hydrolytic enzymes Generation of ROS/RNS - disintegration of membranes, proteins, DNA etc

25. Continuation ., 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 Ca 2+ /ATPase and elevate Ca 2+ & 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 Hazard identification Dose-response assessment Exposure assessment Risk characterization

27. Continuation., 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., Dose-Response Assessment Relationship between the dose and the corresponding response produced. There are two types of dose response relationship Graded dose response relationship : It describes the response of an individual organism to varying dose of chemicals Quantal dose response relationship : characterizes of the distribution of intensity of the effect to different doses in a population of individual organisms.

29. Continuation.,

30. Continuation.,

31. Continuation., 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., 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 Acute toxicity testing : Single dose in one or two different animal species for 14 days Subacute toxicity testing : Repeated dose for upto 90 days. 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). CARCINOGENICITY STUDIES S1A:Need for Carcinogenicity Studies of Pharmaceuticals This document provides a consistent definition of the circumstances under which it is necessary to undertake carcinogenicity studies on new drugs. These recommendations take into account the known risk factors as well as the intended indications and duration of exposure. S1B:Testing for Carcinogenicity of Pharmaceuticals T hi s d oc um e n t p r o v i d e s g ui danc e o n th e ne e d t o carr y o u t carcinogenicity studies in both mice and rats. S1C(R2):Dose Selection for Carcinogenicity Studies of Pharmaceuticals This document 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., GENOTOXICITY STUDIES S2(R1):Guidance on Genotoxicity Testing and Data Interpretation for Pharmaceuticals Intended for Human Use S2A, S2B S2A:Guidance on Specific Aspects of Regulatory Genotoxicity Testsfor Pharmaceuticals This document provided specific guidance and recommendations for in vitro and in vivo tests and on the evaluation of test results. S2B:Genotoxicity: A Standard Battery for Genotoxicity Testing for Pharmaceuticals This document 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 standard battery.

39. Continuation., TOXICOKINETICS ANDPHARMACOKINETICS S3A: Note for Guidance on Toxicokinetics: The Assessment of Systemic Exposure in Toxicity Studies This document gives guidance on developing test strategies in toxicokinetics and the need to integrate pharmacokinetics into toxicitytesting. S3B:Pharmacokinetics: Guidance for Repeated Dose TissueDistribution Studies This document gives guidance on circumstances when repeated dose tissue distribution studies should be considered (i.e., when appropriate data cannot be derived from other sources). TOXICITY TESTING S4:Duration of Chronic Toxicity Testing in Animals (Rodent and Non Rodent Toxicity Testing) The text incorporates the guidance for repeat-dose toxicity tests that was agreed at the time of ICH 1, in 1991 (reduction of the duration of repeat dose toxicity studies in the rat from 12 to 6 months).

40. Continuation., REPRODUCTIVE TOXICOLOGY S5(R2):Detection of Toxicity to Reproduction for Medicinal Products &Toxicity to Male Fertility S5A,S5B(M) This document provides guidance on tests for reproductive toxicity. It defines the periods of treatment to be used in animals to better reflect human exposure to medical products and allow more specific identification of stages at risk. BIOTECHNOLOGICAL PRODUCTS S6(R1):Preclinical Safety Evaluation of Biotechnology-Derived Pharmaceuticals This document covers the pre-clinical safety testing requirements for biotechnological products. PHARMACOLOGY STUDIES S7A: Safety Pharmacology Studies for Human Pharmaceuticals This document addresses the definition, objectives and scope of safety pharmacology studies. S7B: The Non-Clinical Evaluation of the Potential for Delayed Ventricular Repolarization (QT Interval Prolongation) by Human T h i s G u i del i n e d e scr i b e s a n o n - cl i n i c a l t e s ti n g strategy for assessing the potential of a test substance to delay ventricular repolarization.

41. Continuation., IMMUNOTOXICOLOGY STUDIES S8:Immunotoxicity Studies for Human Thi s G uide l i n e addresse s th e r e comm e nd atio n s o n n o n cli n ica l t e st i n g for immunosuppression induced by low molecular weight drugs (non-biologicals). NONCLINICAL EVALUATION FOR ANTICANCER PHARMACEUTICALS S9:Nonclinical Evaluation for Anticancer This Guideline 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-derived pharmaceuticals. PHOTOSAFETY EVALUATION S10:Photosafety Evaluation of Pharmaceuticals (Draft Document) The S10 draft Guideline has been released for consultation under Step 2 of the ICH process in November 2012.

42. 2. OECD - Organisation for Economic Co-operation and Development T he mission of the OECD is to promote policies that will improve 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 solutions to common problems. GUIDELINES: 402 Acute Dermal Toxicity 403 Acute Inhalation Toxicity 404 Acute Dermal Irritation/Corrosion 405 Acute Eye Irritation/Corrosion 406 Skin Sensitisation 407 Repeated Dose 28-day Oral Toxicity Study in Rodents 408 Repeated Dose 90-Day Oral Toxicity Study in Rodents 409 Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents 410 Repeated Dose Dermal Toxicity: 21/28-dayStudy

43. Continuation., 411 Subchronic Dermal Toxicity: 90-day Study 412 Repeated Dose Inhalation Toxicity: 28-day or 14-day Study 413 Subchronic Inhalation Toxicity: 90-day Study 414 Prenatal Developmental Toxicity Study 415 One-Generation Reproduction Toxicity Study 416 Two-Generation Reproduction Toxicity Study 417 Toxicokinetics 420 Acute Oral Toxicity - Fixed DoseMethod 421 Reproduction/Developmental Toxicity Screening Test 422 Combined Repeated Dose Toxicity Study with the Reproduction 423 Acute Oral toxicity - Acute Toxic Class Method 424 Neurotoxicity Study in Rodents 425 Acute Oral Toxicity: Up-and-Down Procedure 451 Carcinogenicity Studies 452 Chronic Toxicity Studies 453 Combined Chronic Toxicity/Carcinogenicity Studies

44. Continuation., 471 Bacterial Reverse Mutation Test 473 In vitro Mammalian Chromosomal AberrationTest 474 Mammalian Erythrocyte Micronucleus Test 475 Mammalian Bone Marrow Chromosomal Aberration Test 476 In vitro Mammalian Cell Gene Mutation Test 477 Genetic Toxicology: Sex-Linked Recessive Lethal Test in Drosophila melanogaster 478 Genetic Toxicology: Rodent Dominant Lethal Test 479 Genetic Toxicology: In vitro Sister Chromatid Exchange Assay in Mammalian Cells 480 Genetic Toxicology: Saccharomyces cerevisiae , Gene Mutation Assay 481 Genetic Toxicology: Saccharomyces cerevisiae , Mitotic Recombination Assay 482 Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells in vitro 483 Mammalian Spermatogonial Chromosome Aberration Test 484 Genetic Toxicology: Mouse Spot Test 485 Genetic Toxicology: Mouse Heritable Translocation Assay 486 Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells invivo

45. Continuation., DRAFT GUIDELINES 403 Acute Inhalation Toxicity 404 Acute Dermal Irritation/Corrosion 405 Acute Eye Irritation/Corrosion 426 Developmental Neurotoxicity Study 429 Skin Sensitisation: Local Lymph Node Assay 430 In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER) 431 In Vitro Skin Corrosion: Human Skin Model Test 432 In Vitro 3T3 NRU Phototoxicity Test

46. 3. EPA - Environment Protection Agencies Guidlines Acute toxicity test guidelines (Group A) Subchronic toxicity test guidelines (Group B) Chronic toxicity test guidelines (Group C) Genetic toxicity test guidelines (Group D) Neurotoxicity test guidelines (Group E) Special study test guidelines (Group F) Health effect chemical specific test guideline (Group G) Supplemental guidance.

47. Continuation., Acute toxicity test guidelines (Group A) 870.1000 - Acute Toxicity Testing--Background (December 2002) 870.1100 - Acute Oral Toxicity (December 2002) 870.1200 - Acute Dermal Toxicity (August 1998) 870.1300 - Acute Inhalation Toxicity (August 1998) 870.2400 - Acute Eye Irritation (August 1998) 870.2500 - Acute Dermal Irritation (August 1998) 870.2600 - Skin Sensitization (March 2003)

48. Continuation., Subchronic toxicity test guidelines (Group B) 870.3050 - Repeated Dose 28-Day Oral Toxicity Study in Rodents (July 2000) 870.3100 - 90-Day Oral Toxicity in Rodents (August 1998) 870.3150 - 90-Day Oral Toxicity in Nonrodents (August 1998) 870.3200 - 21/28-Day Dermal Toxicity (August 1998) 870.3250 - 90-Day Dermal Toxicity (August 1998 870.3465 - 90-Day Inhalation Toxicity (August 1998) 870.3550 - Reproduction/Developmental Toxicity Screening Test (July 2000) 870.3650 - Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (July 2000) 870.3700 - Prenatal Developmental Toxicity Study (August 1998) 870.3800 - Reproduction and Fertility Effects (August 1998)

49. Continuation., Chronic toxicity test guidelines (Group C) 870.4100 - Chronic Toxicity (August 1998) 870.4200 - Carcinogenicity (August 1998) 870.4300 - Combined Chronic Toxicity/Carcinogenicity (August 1998)

50. Continuation., Genetic toxicity test guidelines (Group D) 870.5100 - Bacterial Reverse Mutation Test (August 1998) 870.5195 - Mouse Biochemical Specific Locus Test (August 1998) 870.5200 - Mouse Visible Specific Locus Test (August 1998) 870.5250 - Gene Mutation in Neurospora crassa (August 1998) 870.5275 - Sex-linked Recessive Lethal Test in Drosophila melanogaster (August 1998) 870.5300 - In vitro Mammalian Cell Gene Mutation Test (August 1998) 870.5375 - In vitro Mammalian Chromosome Aberration Test (August 1998) 870.5380 - Mammalian Spermatogonial Chromosomal Aberration Test (August 1998)) 870.5385 - Mammalian Bone Marrow Chromosomal Aberration Test (August 1998) 870.5395 - Mammalian Erythrocyte Micronucleus Test (August 1998) 870.5450 - Rodent Dominant Lethal Assay (August 1998) 870.5460 - Rodent Heritable Translocation Assays (August 1998) 870.5500 - Bacterial DNA Damage or Repair Tests (August 1998) 870.5550 - Unscheduled DNA Synthesis in Mammalian Cells in Culture (August 1998) 870.5575 - Mitotic Gene Conversion in Saccharomyces cerevisiae (August 1998) 870.5900 - In vitro Sister Chromatid Exchange Assay (August 1998) 870.5915 - In vivo Sister Chromatid Exchange Assay (August 1998)

51. Continuation., Neurotoxicity test guidelines (Group E) 870.6100 - Acute and 28-Day Delayed Neurotoxicity of Organophosphorus Substances (August 1998) 870.6200 - Neurotoxicity Screening Battery (August 1998) 870.6300 - Developmental Neurotoxicity Study (August 1998) 870.6500 - Schedule-Controlled Operant Behavior (August 1998) 870.6850 - Peripheral Nerve Function (August 1998) 870.6855 - Neurophysiology Sensory Evoked Potentials (August 1998)

52. Continuation., Health effect chemical specific test guideline (Group G) 870.7200 - Companion Animal Safety (August 1998) 870.7485 - Metabolism and Pharmacokinetics (August 1998) 870.7600 - Dermal Penetration (August 1998) 870.7800 - Immunotoxicity (August 1998) 870.8355 - Combined Chronic Toxicity /Carcinogenicity Testing of Respirable Fibrous Particles (July 2001)

53. OECD Principles of Good Laboratory Practice (GLP)

54. OECD The Organisation for Economic Co-operation and Development (OECD) is an intergovernmental organisation (OECD Secretariat, located in Paris, France) R epresentatives 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 198 1. 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 D ata submitted are a true reflection of the results that are obtained during the study. N ot to indulge in any fraud activity by labs. I nternational acceptance of tests

61. GLP Principles Test Facility Organization and Personnel Quality Assurance Program Facilities Apparatus, Material, and Reagents Test Systems Test and Reference Items SOPs Performance of the Study Reporting of Study Results 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 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 e nsure 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 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 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. 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 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. Responsibilities of the Quality Assurance Personnel To m aintain 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 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. 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 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 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. 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 Physical/Chemical Apparatus used for the generation of physical/chemical data should be suitably located and of appropriate design and adequate capacity. 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 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 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 that b 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 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. 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 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 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. 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 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 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 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 . LD 50 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 (LD 50 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.

81. Reference 2020 Springer Nature Switzerland AG. Google Search Wikipedia.org Slideshare.net

82. Quote ~ ” Your limitation—it’s only your imagination ”

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