Alternative methods to animal toxicity testing

alternative method to animal toxicity testing

1. Institute of Pharmaceutical Science Kurukshetra University Presentation on:Alternative method to animal Toxicity Testing. Presented By : Sachin sharma M.Pharma 2 nd Sem Pharmacology Roll No: 5461076 1

2. Alternative Methods To Animal Toxicity Testing

3. CONTENT Introduction Need for alternatives to animals The 3Rs Principles Validation of alternative methods Alternative methods HET- CAM Method 3

4. Introduction Alternative methods to animals testing are the development and implementation of test method that avoid use of live animals or use of less animals in method. The council directive on protection of animals used for experiments and scientific purpose in article 23 “The commission and member states should encourage research into development and validation of alternative methods which could provide the same level of information as that obtained in experiment using animals but which involves less animal”. Alternative methods able to do: Reduce Refine Replace collectively called as “The 3Rs Principle”. 4

5. Needs for alternative methods Because in laboratory animals may be: Poisoned. Deprived of food water and sleep. Applied with skin and eye irritants. Subjected to psychological stress. Deliberately infected with infected disease. 5

6. The 3Rs Principle The 3Rs principles were defined in 1959 by W.M.S Russel and R.L Bruch in subject “The Principle of Humane experimental technique”. They provide a strategy for rational and stepwise approach to minimising animals use and suffering in experiments without compromising the quality and quantity of scientific work being undertaken. 6

7. Reduction means any decrease in numbers of animals used to obtain information of a given amount and precision. Refinement means any decrease in the incidence or severity of procedures applied to animals. Replacement means the substitution of conscious living animals with non sentient. 7

8. Validation of alternative methods European centre for validation of alternative method (ECVAM) was established in 1991. ECVAM serves as information centre and to maintain a database on in vitro test and validated method. “Test validation is a process based on principle by which the reliability and relevance of particular test/method are established for specific purpose”. Validation process carry out by three organisation Organisation for economic corporation and development (OECD). European centre for validation of alternative methods(ECVAM). Interagency co-ordinating committee on validation of alternative method. 8

9. There are two type of validation studies. Validation studies Prospective Retrospective 9

10. Prospective validation studies Involve generation of new data. Composed of six stages. Research Method Development 10

11. Pre-validation Validation Review Regulatory Acceptance 11

12. Alternative Method In-vitro test Toxicity test Corrosivity test Insilico Mathematical model Computer simulation 12

13. Genotoxicity Test Ames Test Occular toxicity Test HET-CAM Test Hepatotoxicity Test Natural Red Uptake test Corrosivity Test Transcutaneous Electrical Resistance Test Method HET-CAM: Hen’s egg test- chorio allantoic membrane Toxicity Test 13

14. HET-CAM TEST The fresh fertile white leghorn eggs are used. The eggs are held in optimized incubation condition. On day 10 inner egg membrane is removed , after careful removal the living vascular chorioallantoic membrane is exposed. The test substance is droped over CAM in a volume of 0.2-0.3 ml and irrigated after 20 sec. with 5ml warm water. The CAM, the blood vessels, including capillary system, and albumin are examined and scored for irritant effects 0.5, 2, 5min after test compound application. 14

15. 0.5(min) 2(min) 5(min) haemorrhage 5 3 1 Coagulation 9 7 5 Effects Score Scoring Scheme Classification scheme Cumulative score Irritation Assessment Up to 0.9 Practically none 1 to 4.9 Slight 5 to 8.9 Moderate 9 and above Strong 15

16. Thank You 16