Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursu... (32008R0440)
INHALT
Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) (Text with EEA relevance)
- COUNCIL REGULATION (EC)
- No 440/2008
- of 30 May 2008
- laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)
- (Text with EEA relevance)
- Article 1
- Article 2
- Article 3
- Article 4
- ANNEX
- PART A: METHODS FOR THE DETERMINATION OF PHYSICO-CHEMICAL PROPERTIES
- A.1. MELTING/FREEZING TEMPERATURE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.4.1. Capillary method
- 1.4.1.1. Melting temperature devices with liquid bath
- 1.4.1.2. Melting temperature devices with metal block
- 1.4.1.3. Photocell detection
- 1.4.2. Hot stages
- 1.4.2.1. Kofler hot bar
- 1.4.2.2. Melt microscope
- 1.4.2.3. Meniscus method
- 1.4.3. Method to determine the freezing temperature
- 1.4.4. Thermal analysis
- 1.4.4.1 Differential thermal analysis (DTA)
- 1.4.4.2 Differential scanning calorimetry (DSC)
- 1.4.5. Pour point
- 1.5. QUALITY CRITERIA
- TABLE: APPLICABILITY OF THE METHODS
- A.
- Capillary methods
- B.
- Hot stages and freezing methods
- C.
- Thermal analysis
- D.
- Pour point
- 1.6. DESCRIPTION OF THE METHODS
- 1.6.1. Methods with capillary tube
- Figure 1
- 1.6.1.1. Melting temperature devices with liquid bath apparatus
- Figure 2
- Bath liquid:
- Thermometer:
- Procedure:
- Calculation:
- 1.6.1.2. Melting temperature devices with metal block
- Apparatus:
- Thermometer:
- Figure 3
- 1.6.1.3. Photocell detection
- 1.6.2. Hot stages
- 1.6.2.1. Kofler hot bar
- 1.6.2.2. Melt microscope
- 1.6.2.3. Meniscus method (polyamides)
- 1.6.3. Methods for the determination of the freezing temperature
- 1.6.4. Thermal analysis
- 1.6.4.1. Differential thermal analysis
- 1.6.4.2. Differential scanning calorimetry
- 1.6.5. Determination of the pour point
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Appendix
- For additional technical details, the following standards may be consulted for example.
- 1. Capillary methods
- 1.1. Melting temperature devices with liquid bath
- 1.2. Melting temperature devices with metal block
- 2. Hot stages
- 2.1. Kofler hot bar
- 2.2. Melt microscope
- 2.3. Meniscus method (polyamides)
- 3. Methods for the determination of the freezing temperature
- 4. Thermal analysis
- 4.1. Differential thermal analysis
- 4.2. Differential scanning calorimetry
- 5. Determination of the pour point
- A.2. BOILING TEMPERATURE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.4.1. Determination by use of the ebulliometer
- 1.4.2. Dynamic method
- 1.4.3. Distillation method for boiling temperature
- 1.4.4. Method according to Siwoloboff
- 1.4.5. Photocell detection
- 1.4.6. Differential thermal analysis
- 1.4.7. Differential scanning calorimetry
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHODS
- 1.6.1. Ebulliometer
- 1.6.2. Dynamic method
- 1.6.3. Distillation process (boiling range)
- 1.6.4. Method according to Siwoloboff
- Figure 1
- 1.6.5. Photocell detection
- 1.6.6. Thermal analysis
- 1.6.6.1. Differential thermal analysis
- 1.6.6.2. Differential scanning calorimetry
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Appendix
- For additional technical details, the following standards may be consulted for example.
- 1. Ebulliometer
- 2. Distillation process (boiling range)
- 3. Differential thermal analysis and differential scanning calorimetry
- A.3. RELATIVE DENSITY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES (1) (3)
- 1.4. PRINCIPLE OF THE METHODS
- 1.4.1. Buoyancy methods
- 1.4.1.1. Hydrometer
- (for liquid substances)
- 1.4.1.2. Hydrostatic balance
- (for liquid and solid substances)
- 1.4.1.3. Immersed body method
- (for liquid substances) (4)
- 1.4.2. Pycnometer methods
- 1.4.3. Air comparison pycnometer
- (for solids)
- 1.4.4. Oscillating densitimeter
- (5) (6) (7)
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHODS
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Appendix
- For additional technical details, the following standards may be consulted for example.
- 1. Buoyancy methods
- 1.1. Hydrometer
- 1.2. Hydrostatic balance
- For solid substances
- For liquid substances
- 1.3. Immersed body method
- 2. Pycnometer methods
- 2.1. For liquid substances
- 2.2. For solid substances
- 3. Air comparison pycnometer
- A.4. VAPOUR PRESSURE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHODS
- 1.4.1. Dynamic method
- 1.4.2. Static method
- 1.4.3. Isoteniscope
- 1.4.4. Effusion method: Vapour pressure balance
- 1.4.5. Effusion method: By loss of weight or by trapping vaporisate
- 1.4.6. Gas saturation method
- 1.4.7. Spinning rotor
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHODS
- 1.6.1. Dynamic measurement
- 1.6.1.1. Apparatus
- 1.6.1.2. Measurement procedure
- 1.6.2. Static measurement
- 1.6.2.1. Apparatus
- 1.6.2.2. Measurement procedure
- 1.6.3. Isoteniscope
- 1.6.4. Effusion method: vapour pressure balance
- 1.6.4.1. Apparatus
- 1.6.4.2. Measurement procedure
- 1.6.5. Effusion method — by loss of weight
- 1.6.5.1. Apparatus
- 1.6.5.2. Measurement procedure
- 1.6.6. Gas saturation method
- 1.6.6.1. Apparatus
- Inert gas:
- Flow control:
- Traps to collect vapour:
- Heat exchanger:
- Saturator column:
- 1.6.6.2. Measurement procedure
- Preparation of the saturator column:
- Measurement:
- 1.6.6.3. Calculation of vapour pressure
- 1.6.7. Spinning rotor (8, 11, 13)
- 1.6.7.1. Apparatus
- 1.6.7.2. Measurement procedure
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Appendix 1
- Estimation method
- INTRODUCTION
- ESTIMATION METHOD
- CALCULATION PROCEDURE
- REPORT
- LITERATURE
- Appendix 2
- Figure 1
- Apparatus for determining the vapour pressure curve according to the dynamic method
- Figure 2a
- Apparatus for determining the vapour pressure curve according to the static method (using a U-tube manometer)
- Figure 2b
- Apparatus for determining the vapour pressure curve according to the static method (using a pressure indicator)
- Figure 3
- Isoteniscope (see reference 7)
- Figure 4
- Apparatus for determining the vapour pressure curve according to the vapour pressure balance method
- Figure 5
- Example of apparatus for evaporation at low pressure by effusion methode, with an effusion cell volume of 8 cm
- 3
- Figure 6a
- An example of a flow system for the determination of vapour pressure by the gas saturation method
- Figure 6b
- An example of system for the determination of vapour pressure by the gas saturation method, with a capillary placed after the saturation chamber
- Figure 7
- Example of the experimental set-up for spinning rotor method
- Figure 8
- Example of spinning rotor measuring head
- A.5. SURFACE TENSION
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHODS
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHODS
- 1.6.1. Plate method
- 1.6.2. Stirrup method
- 1.6.3. Ring method
- 1.6.4. OECD harmonised ring method
- 1.6.4.1. Apparatus
- 1.6.4.1.1.
- Mobile sample table
- 1.6.4.1.2.
- Force measuring system
- 1.6.4.1.3.
- Measuring body (ring)
- Figure
- Measuring body
- 1.6.4.1.4.
- Measurement vessel
- 1.6.4.2. Preparation of the apparatus
- 1.6.4.2.1.
- Cleaning
- 1.6.4.2.2.
- Calibration of the apparatus
- Mounting:
- Zero point adjustment:
- Calibrations:
- 1.6.4.3. Preparation of samples
- 1.6.5. Test conditions
- 1.6.6. Performance of test
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF RESULTS
- 4. REFERENCES
- A.6. WATER SOLUBILITY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.4.1. Column elution method
- 1.4.2. Flask method
- 1.5. QUALITY CRITERIA
- 1.5.1. Repeatability
- 1.5.2. Sensitivity
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Test conditions
- 1.6.2. Preliminary test
- 1.6.3. Column elution method
- 1.6.3.1. Support material, solvent and eluent
- 1.6.3.2. Loading of the support
- Test procedure:
- 1.6.3.3. Column elution method with recirculating pump
- Apparatus
- Measurement procedure
- 1.6.3.4. Column elution method with levelling vessel
- Apparatus
- (see figures 4 and 3)
- Measurement procedure
- 1.6.4. Flask method
- 1.6.4.1. Apparatus
- 1.6.4.2. Measurement procedure
- 1.6.5. Analysis
- 2. DATA
- 2.1. COLUMN ELUTION METHOD
- 2.2. FLASK METHOD
- 3. REPORTING
- 3.1. COLUMN ELUTION METHOD
- 3.2. FLASK METHOD
- 4. REFERENCES
- Appendix
- Figure 1
- Column elution method with recirculating pump
- Figure 2
- A typical microcolumn
- Figure 3
- A typical microcolumn
- Figure 4
- Column elution method with levelling vessel
- A.8. PARTITION COEFFICIENT
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCES
- Shake-flask method
- HPLC method
- 1.4. PRINCIPLE OF THE METHOD
- 1.4.1. Shake-flask method
- 1.4.2. HPLC method
- 1.5. QUALITY CRITERIA
- 1.5.1. Repeatability
- Shake-flask method
- HPLC method
- 1.5.2. Sensitivity
- Shake-flask method
- HPLC method
- 1.5.3. Specificity
- Shake-flask method
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Preliminary estimate of the partition coefficient
- 1.6.2. Shake-flask method
- 1.6.2.1. Preparation
- 1.6.2.1.1.
- Pre-saturation of the solvents
- 1.6.2.1.2.
- Preparation for the test
- 1.6.2.1.3.
- Test substance
- 1.6.2.2. Test conditions
- 1.6.2.3. Measurement procedure
- 1.6.2.3.1.
- Establishment of the partition equilibrium
- 1.6.2.3.2.
- Phase separation
- 1.6.2.4. Analysis
- 1.6.3. HPLC method
- 1.6.3.1. Preparation
- Apparatus
- Mobile phase
- Solutes
- Test conditions
- 1.6.3.2.
- Measurement
- Calculation of dead time t
- o
- Calibration graph
- Determination of the capacity factor of the test substance
- 2. DATA
- Shake-flask method
- 3. REPORTING
- For shake-flask method:
- For HPLC method:
- 4. REFERENCES
- Appendix 1
- Calculation/estimation methods
- INTRODUCTION
- ESTIMATION METHOD
- Preliminary estimate of the partition coefficient
- CALCULATION METHODS
- Principle of the calculation methods
- Quality criteria
- Calculation procedures
- Hansch π-method
- Rekker method
- Hansch-Leo method
- Combined method
- Remarks
- Report
- LITERATURE
- Appendix 2
- Recommended Reference Substances for the HLPC Method
- A.9. FLASH-POINT
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.5.1. Repeatability
- 1.5.2. Sensitivity
- 1.5.3. Specificity
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.3. Performance of the test
- 1.6.3.1. Equilibrium method
- 1.6.3.2. Non-equilibrium method
- Abel apparatus:
- Abel-Pensky apparatus:
- Tag apparatus:
- Pensky-Martens apparatus:
- Remarks:
- 3. REPORTING
- 4. REFERENCES
- A.10. FLAMMABILITY (SOLIDS)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF METHOD
- 1.6.1. Preliminary screening test
- 1.6.2. Burning rate test
- 1.6.2.1. Preparation
- 1.6.2.2. Test conditions
- 1.6.2.3. Performance of the test
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF THE RESULT
- 4. REFERENCES
- Appendix
- Figure
- Mould and accessories for the preparation of the pile
- A.11. FLAMMABILITY (GASES)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Apparatus
- 1.6.2. Test conditions
- 1.6.3. Performance of the test
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- A.12. FLAMMABILITY (CONTACT WITH WATER)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. PRINCIPLE OF THE METHOD
- 1.3.1. Step 1
- 1.3.2. Step 2
- 1.3.3. Step 3
- 1.3.4. Step 4
- 1.4. REFERENCE SUBSTANCES
- 1.5. QUALITY CR1TERIA
- 1.6. DESCRIPTION OF METHODS
- 1.6.1. Step 1
- 1.6.1.1. Test conditions
- 1.6.1.2. Performance of the test
- 1.6.2. Step 2
- 1.6.2.1. Apparatus
- 1.6.2.2. Test conditions
- 1.6.2.3. Performance of the test
- 1.6.3. Step 3
- 1.6.3.1. Test conditions
- 1.6.3.2. Performance of the test
- 1.6.4. Step 4
- 1.6.4.1. Apparatus
- 1.6.4.2. Test conditions
- 1.6.4.3. Performance of the test
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Appendix
- Figure
- Apparatus
- A.13. PYROPHORIC PROPERTIES OF SOLIDS AND LIQUIDS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Apparatus
- 1.6.2. Performance of the test
- (a) Powdery solids
- (b) liquids
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION
- 3. REPORTING
- 4. REFERENCES
- A.14. EXPLOSIVE PROPERTIES
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.4.1. Safety-in-handling tests (3)
- 1.4.2. Thermal sensitivity
- 1.4.3. Mechanical sensitivity (shock)
- 1.4.4. Mechanical sensitivity (friction)
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF METHOD
- 1.6.1. Thermal sensitivity (effect of a flame)
- 1.6.1.1. Apparatus
- 1.6.1.2. Test conditions
- 1.6.1.3. Performance of the tests
- 1.6.1.4. Evaluation
- 1.6.2. Mechanical sensitivity (shock)
- 1.6.2.1. Apparatus (figure 4)
- 1.6.2.2. Test conditions
- 1.6.2.3. Performance of the tests
- 1.6.2.4. Evaluation
- 1.6.3. Mechanical sensitivity (friction)
- 1.6.3.1. Apparatus (figure 5)
- 1.6.3.2. Test conditions
- 1.6.3.3. Performance of the tests
- 1.6.3.4. Evaluation
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION AND EVALUATION OF RESULTS
- 4. REFERENCES
- Appendix
- Example of material specification for thermal sensitivity test (see DIN 1623)
- Figure 1
- Thermal sensitivity test apparatus
- Figure 2
- Thermal sensitivity test
- Figure 3
- Heating rate calibration for thermal sensitivity test
- Figure 4
- Shock test apparatus
- Figure 4
- Continued
- Figure 5
- Friction sensitivity apparatus
- A.15. AUTO-IGNITION TEMPERATURE (LIQUIDS AND GASES)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Apparatus
- 1.6.2. Test conditions
- 1.6.3. Performance of the test
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- A.16. RELATIVE SELF-IGNITION TEMPERATURE FOR SOLIDS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCE
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Apparatus
- 1.6.1.1. Oven
- 1.6.1.2. Wire mesh cube
- 1.6.1.3. Thermocouples
- 1.6.1.4. Recorder
- 1.6.2. Test conditions
- 1.6.3. Performance of the test
- 2. DATA
- 3. REPORTING
- 4. REFERENCES
- Figure 1
- Pattern of 20 mm test cube
- Figure 2
- Typical temperature/time curve
- A.17. OXIDISING PROPERTIES (SOLIDS)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCE
- 1.4. PRINCIPLE OF THE METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Preparation
- 1.6.1.1. Test substance
- 1.6.1.2. Combustible substance
- 1.6.1.3. Ignition source
- 1.6.2. Performance of the test
- 1.6.2.1. Preliminary test
- 1.6.2.2. Train test
- 2. DATA
- 3. REPORT
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF THE RESULT
- 4. REFERENCES
- Appendix
- Figure
- Mould and accessories for the preparations of the pile
- A.18. NUMBER-AVERAGE MOLECULAR WEIGHT AND MOLECULAR WEIGHT DISTRIBUTION OF POLYMERS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparation of the standard polystyrene solutions
- 1.6.2. Preparation of the sample solution
- 1.6.3. Apparatus
- 1.6.4. Injection and solvent delivery system
- 1.6.5. Column
- 1.6.6. Theoretical plates
- 1.6.7. Separation efficiency
- 1.6.8. Solvents
- 1.6.9. Temperature control
- 1.6.10. Detector
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Instrumentation:
- 2.2.3. Calibration of the system:
- 2.2.4. Evaluation:
- 3. REFERENCES
- Appendix
- Examples of other methods for determination of number average molecular weight (Mn) for polymers
- 1. Use of colligative properties
- 2. End-group analysis
- 3. References
- A.19. LOW MOLECULAR WEIGHT CONTENT OF POLYMERS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparation of the standard polystyrene solutions
- 1.6.2. Preparation of the sample solution
- 1.6.3. Correction for content of impurities and additives
- 1.6.4. Apparatus
- 1.6.5. Injection and solvent delivery system
- 1.6.6. Column
- 1.6.7. Theoretical plates
- 1.6.8. Separation efficiency
- 1.6.9. Solvents
- 1.6.10. Temperature control
- 1.6.11. Detector
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Instrumentation:
- 2.2.3. Calibration of the system:
- 2.2.4. Information on the low molecular weight polymer content:
- 2.2.5. Evaluation:
- 3. REFERENCES
- Appendix
- Guidance for correcting low molecular content for the presence of insoluble polymer
- A.20. SOLUTION/EXTRACTION BEHAVIOUR OF POLYMERS IN WATER
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. REFERENCE SUBSTANCES
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. QUALITY CRITERIA
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Equipment
- 1.5.2. Sample preparation
- 1.5.3. Procedure
- 1.5.4. Analysis
- 1.5.4.1. Test conducted with one sample size
- 1.5.4.2. Test conducted with two different sample sizes
- 2. DATA
- 2.1. TEST CONDUCTED WITH ONE SAMPLE SIZE
- 2.2. TEST CONDUCTED WITH TWO DIFFERENT SAMPLE SIZES
- 3. REPORTING
- 3.1. TEST REPORT
- 3.1.1. Test substance:
- 3.1.2. Experimental conditions:
- 3.1.3. Results:
- 4. REFERENCES
- A.21. OXIDISING PROPERTIES (LIQUIDS)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCE
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE METHOD
- 1.6.1. Preparation
- 1.6.1.1. Combustible substance
- 1.6.1.2. Apparatus
- 1.6.1.2.1.
- Pressure vessel
- 1.6.1.2.2.
- Ignition system
- 1.6.2. Performance of the test
- (15)
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 3. REPORT
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF THE RESULTS(22)
- 4. REFERENCES
- Figure 1
- Pressure vessel
- Figure 2
- Support stand
- Figure 3
- Ignition system
- PART B: METHODS FOR THE DETERMINATION OF TOXICITY AND OTHER HEALTH EFFECTS
- GENERAL INTRODUCTION
- A. CHARACTERISATION OF THE TEST SUBSTANCE
- B. ANIMAL CARE
- (i) Housing conditions
- (ii) Feeding conditions
- C. ALTERNATIVE TESTING
- D. EVALUATION AND INTERPRETATION
- E. LITERATURE REFERENCES
- B.1 bis. ACUTE ORAL TOXICITY — FIXED DOSE PROCEDURE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Selection of animal species
- 1.4.2. Housing and feeding conditions
- 1.4.3. Preparation of animals
- 1.4.4. Preparation of doses
- 1.5. PROCEDURE
- 1.5.1. Administration of doses
- 1.5.2. Sighting study
- 1.5.3. Main study
- 1.5.3.1. Numbers of animals and dose levels
- 1.5.3.2. Limit test
- 1.6. OBSERVATIONS
- 1.6.1. Body weight
- 1.6.2. Pathology
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Appendix 1
- FLOW CHART FOR THE SIGHTING STUDY
- Appendix 2
- FLOW CHART FOR THE MAIN STUDY
- Appendix 3
- CRITERIA FOR CLASSIFICATION OF TEST SUBSTANCES WITH EXPECTED LD
- 50
- VALUES EXCEEDING 2 000 MG/KG WITHOUT THE NEED FOR TESTING.
- TESTING AT DOSES ABOVE 2 000 MG/KG
- Sighting study
- Main study
- Appendix 4
- TEST METHOD B.1 bis
- Guidance on classification according to the EU scheme to cover the transition period until full implementation of the Globally Harmonised Classification System (GHS) (taken from reference (8))
- B.1 tris. ACUTE ORAL TOXICITY — ACUTE TOXIC CLASS METHOD
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST
- 1.4. DESCRIPTION OF THE METHOD
- 1.4.1. Selection of animal species
- 1.4.2. Housing and feeding conditions
- 1.4.3. Preparation of animals
- 1.4.4. Preparation of doses
- 1.5. PROCEDURE
- 1.5.1. Administration of doses
- 1.5.2. Number of animals and dose levels
- 1.5.3. Limit test
- 1.6. OBSERVATIONS
- 1.6.1. Body weight
- 1.6.2. Pathology
- 2. DATA
- 3. REPORTING
- 3.1. Test report
- 4. REFERENCES
- Appendix 1
- PROCEDURE TO BE FOLLOWED FOR EACH OF THE STARTING DOSES
- GENERAL REMARKS
- Appendix 1A
- TEST PROCEDURE WITH A STARTING DOSE OF 5 MG/KG BODY WEIGHT
- Appendix 1B
- TEST PROCEDURE WITH A STARTING DOSE OF 50 MG/KG BODY WEIGHT
- Appendix1C
- TEST PROCEDURE WITH A STARTING DOSE OF 300 MG/KG BODY WEIGHT
- Appendix 1D
- TEST PROCEDURE WITH A STARTING DOSE OF 2 000 MG/KG BODY WEIGHT
- Appendix 2
- CRITERIA FOR CLASSIFICATION OF TEST SUBSTANCES WITH EXPECTED LD
- 50
- VALUES EXCEEDING 2 000 MG/KG WITHOUT THE NEED FOR TESTING
- TESTING AT DOSES ABOVE 2 000 MG/KG
- Appendix 3
- TEST METHOD B.1 tris: Guidance on classififcation according to EU scheme to cover the transition period until full implementation of the Globally Harmonised Classification System (GHS) (taken from reference (8))
- B.2. ACUTE TOXICITY (INHALATION)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.2.1. Experimental animals
- 1.6.2.2. Number and sex
- 1.6.2.3. Exposure concentrations
- 1.6.2.4. Limit test
- 1.6.2.5. Exposure time
- 1.6.2.6. Equipment
- 1.6.2.7. Observation period
- 1.6.3. Procedure
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.3. ACUTE TOXICITY (DERMAL)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.2.1. Experimental animals
- 1.6.2.2. Number and sex
- 1.6.2.3. Dose levels
- 1.6.2.4. Limit test
- 1.6.2.5. Observation period
- 1.6.3. Procedure
- Assessment of toxicity in the other sex
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.4. ACUTE TOXICITY: DERMAL IRRITATION/CORROSION
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparation for the
- in vivo
- test
- 1.4.1.1. Selection of animal species
- 1.4.1.2. Preparation of the animals
- 1.4.1.3. Housing and feeding conditions
- 1.4.2. Test procedure
- 1.4.2.1. Application of the test substance
- 1.4.2.2. Dose level
- 1.4.2.3. Initial test (
- in vivo
- dermal irritation/corrosion test using one animal)
- 1.4.2.4. Confirmatory test (
- in vivo
- dermal irritation test with additional animals)
- 1.4.2.5. Observation period
- 1.4.2.6. Clinical observations and grading of skin reactions
- 2. DATA
- 2.1. PRESENTATION OF RESULTS
- 2.2. EVALUATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Table I
- GRADING OF SKIN REACTIONS
- Appendix
- A Sequential Testing Strategy for Dermal Irritation and Corrosion
- GENERAL CONSIDERATIONS
- DESCRIPTION OF THE EVALUATION AND TESTING STRATEGY
- REFERENCES
- Figure
- TESTING AND EVALUATION STRATEGY FOR DERMAL IRRITATION/CORROSION
- B.5. ACUTE TOXICITY: EYE IRRITATION/CORROSION
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparation for the
- in vivo
- test
- 1.4.1.1. Selection of species
- 1.4.1.2. Preparation of animals
- 1.4.1.3. Housing and feeding conditions
- 1.4.2. Test procedure
- 1.4.2.1. Application of the test substance
- 1.4.2.2. Irrigation
- 1.4.2.3. Dose level
- 1.4.2.3.1.
- Testing of liquids
- 1.4.2.3.2.
- Testing of solids
- 1.4.2.3.3.
- Testing of aerosols
- 1.4.2.4. Initial test (
- in vivo
- eye irritation/corrosion test using one animal)
- 1.4.2.5. Local anaesthetics
- 1.4.2.6. Confirmatory test (
- in vivo
- eye irritation test with additional animals)
- 1.4.2.7. Observation period
- 1.4.2.7.1.
- Clinical observations and grading of eye reactions
- 2. DATA
- 2.2. EVALUATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF THE RESULTS
- 4. REFERENCES
- Table I
- GRADING OF OCULAR LESIONS
- Cornea
- Iris
- Conjunctivae
- Chemosis
- Appendix
- A Sequential Testing Strategy for Eye Irritation and Corrosion
- GENERAL CONSIDERATIONS
- DESCRIPTION OF THE STEPWISE TESTING STRATEGY
- REFERENCES
- Figure
- TESTING AND EVALUATION STRATEGY FOR EYE IRRITATION/CORROSION
- B.6. SKIN SENSITISATION
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. DESCRIPTION OF THE TEST METHODS
- 1.5.1.
- Guinea-Pig Maximisation Test (GPMT)
- 1.5.1.1.
- Preparations
- 1.5.1.2.
- Test conditions
- 1.5.1.2.1. Test animals
- 1.5.1.2.2. Number and sex
- 1.5.1.2.3. Dose levels
- 1.5.1.3.
- Procedure
- 1.5.1.3.1. Induction
- 1.5.1.3.2. Challenge
- 1.5.1.3.3. Observation and Grading: treated and control groups
- 1.5.2.
- Buehler test
- 1.5.2.1.
- Preparations
- 1.5.2.2.
- Test conditions
- 1.5.2.2.1. Test animals
- 1.5.2.2.2. Number and sex
- 1.5.2.2.3. Dose levels
- 1.5.2.3.
- Procedure
- 1.5.2.3.1. Induction
- 1.5.2.3.2. Challenge
- 1.5.2.3.3. Observation and grading
- 2. DATA (GPMT and Buehler test)
- 3. REPORTING (GPMT and Buehler test)
- Test report (GMPT and Buehler test)
- 4. REFERENCES
- Appendix
- TABLE
- Magnusson/Kligman grading scale for the evaluation of challenge patch test reactions
- B.7. REPEATED DOSE (28 DAYS) TOXICITY (ORAL)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.2. Test conditions
- 1.4.2.1. Test animals
- 1.4.2.2. Number and sex
- 1.4.2.3. Dose levels
- 1.4.2.4. Limit test
- 1.4.2.5. Observation period
- 1.4.3. Procedure
- 1.4.3.1. General observations
- 1.4.3.2. Body weight and food/water consumption
- 1.4.3.3. Haematology
- 1.4.3.4. Clinical biochemistry
- 1.4.3.5. Gross necropsy
- 1.4.3.6. Histopathological examination
- 2. DATA
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.8. REPEATED DOSE (28 DAYS) TOXICITY (INHALATION)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.2.1. Experimental animals
- 1.6.2.2. Number and sex
- 1.6.2.3. Exposure concentration
- 1.6.2.4. Exposure time
- 1.6.2.5. Equipment
- 1.6.2.6. Observation period
- 1.6.3. Procedure
- 1.6.3.1. Gross necropsy
- 1.6.3.2. Histopathological examination
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.9. REPEATED DOSE (28 DAYS) TOXICITY (DERMAL)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.2.1. Experimental animals
- 1.6.2.2. Number and sex
- 1.6.2.3. Dose levels
- 1.6.2.4. Limit test
- 1.6.2.5. Observation period
- 1.6.3. Procedure
- 1.6.4. Gross necropsy
- 1.6.5. Histopathological examination
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.10. MUTAGENICITY —
- IN VITRO
- MAMMALIAN CHROMOSOME ABERRATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Cells
- 1.4.1.2. Media and culture conditions
- 1.4.1.3. Preparation of cultures
- 1.4.1.4. Metabolic activation
- 1.4.1.5. Test substance/Preparation
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/vehicle
- 1.4.2.2. Exposure concentrations
- 1.4.2.3. Negative and positive controls
- 1.4.3. Procedure
- 1.4.3.1. Treatment with the test substance
- 1.4.3.3. Culture harvest time
- 1.4.3.4. Chromosome preparation
- 1.4.3.5. Analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.11. MUTAGENICITY —
- IN VIVO
- MAMMALIAN BONE MARROW CHROMOSOME ABERRATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Selection of animal species
- 1.4.1.2. Housing and feeding conditions
- 1.4.1.3. Preparation of the animals
- 1.4.1.4. Preparation of doses
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/Vehicle
- 1.4.2.2. Controls
- 1.5. PROCEDURE
- 1.5.1. Number and sex of animals
- 1.5.2. Treatment schedule
- 1.5.3. Dose levels
- 1.5.4. Limit test
- 1.5.5. Administration of doses
- 1.5.6. Chromosome preparation
- 1.5.7. Analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.12. MUTAGENICITY —
- IN VIVO
- MAMMALIAN ERYTHROCYTE MICRONUCLEUS TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Selection of animal species
- 1.4.1.2. Housing and feeding conditions
- 1.4.1.3. Preparation of the animals
- 1.4.1.4. Preparation of doses
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/Vehicle
- 1.4.2.2. Controls
- 1.5. PROCEDURE
- 1.5.1. Number and sex of animals
- 1.5.2. Treatment schedule
- 1.5.3. Dose levels
- 1.5.4. Limit test
- 1.5.5. Administration of doses
- 1.5.6. Bone marrow/blood preparation
- 1.5.7. Analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.13/14. MUTAGENICITY: REVERSE MUTATION TEST USING BACTERIA
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. INITIAL CONSIDERATIONS
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Preparations
- 1.5.1.1. Bacteria
- 1.5.1.2. Medium
- 1.5.1.3. Metabolic activation
- 1.5.1.4. Test substance/Preparation
- 1.5.2. Test conditions
- 1.5.2.1. Test strains (see 1.5.1.1)
- 1.5.2.2. Exposure concentration
- 1.5.2.3. Negative and positive controls
- 1.5.3. Procedure
- 1.5.4. Incubation
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.15. MUTAGENICITY TESTING AND SCREENING FOR CARCINOGENICITY GENE MUTATION —
- SACCHAROMYCES CEREVISIAE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Metabolic activation
- Test conditions
- Tester strains
- Media
- Use of negative and positive controls
- Exposure concentration
- Incubation conditions
- Spontaneous mutation frequencies
- Number of replicates
- Procedure
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.16. MITOTIC RECOMBINATION —
- SACCHAROMYCES CEREVISIAE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Metabolic activation
- Test conditions
- Tester strains
- Media
- Use of negative and positive controls
- Exposure concentrations
- Incubation conditions
- Spontaneous mitotic recombination frequencies
- Number of replicates
- Procedures
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.17. MUTAGENICITY —
- IN VITRO
- MAMMALIAN CELL GENE MUTATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Cells
- 1.4.1.2. Media and culture conditions
- 1.4.1.3. Preparation of cultures
- 1.4.1.4. Metabolic activation
- 1.4.1.5. Test substance/Preparation
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/Vehicle
- 1.4.2.2. Exposure concentrations
- 1.4.2.3. Controls
- 1.4.3. Procedure
- 1.4.3.1. Treatment with the test substance
- 1.4.3.2. Measurement of survival, viability and mutant frequency
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.18. DNA DAMAGE AND REPAIR — UNSCHEDULED DNA SYNTHESIS — MAMMALIAN CELLS
- IN VITRO
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Test conditions
- Number of cultures
- Use of negative and positive controls
- Exposure concentrations
- Cells
- Metabolic activation
- Procedure
- Preparation of cultures
- Treatment of the cultures with the test substance
- Primary rat hepatocytes
- Established cell lines and lymphocytes
- Analysis
- Autoradiographic determinations
- LSC determinations
- 2. DATA
- 2.1. AUTORADIOGRAPHIC DETERMINATIONS
- 2.2. LSC DETERMINATIONS
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. VALUATION AND INTERPRETATION
- 4. REFERENCES
- B.19. SISTER CHROMATID EXCHANGE ASSAY
- IN VITRO
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Preparations
- 1.6.2.
- Test conditions
- Number of cultures
- Use of negative and positive controls
- Exposure concentrations
- 1.6.3.
- Procedure
- Preparation of cultures
- Treatment
- Harvesting of cells
- Chromosome preparation and staining
- Analysis
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.20. SEX-LINKED RECESSIVE LETHAL TEST IN
- DROSOPHILA MELANOGASTER
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLES OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Stocks
- Test substance
- Number of animals
- Route of administration
- Use of negative and positive controls
- Exposure levels
- Procedure
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.21.
- IN VITRO
- MAMMALIAN CELL TRANSFORMATION TESTS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Cells
- Medium
- Test substance
- Metabolic activation
- Test conditions
- Use of negative and positive controls
- Exposure concentrations
- Procedure
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.22. RODENT DOMINANT LETHAL TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Test conditions
- Route of administration
- Experimental animals
- Number and sex
- Use of negative and positive controls
- Dose levels
- Limit test
- Procedure
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.23. MAMMALIAN SPERMATOGONIAL CHROMOSOME ABERRATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Selection of animal species
- 1.4.1.2. Housing and feeding conditions
- 1.4.1.3. Preparation of the animals
- 1.4.1.4. Preparation of doses
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/Vehicle
- 1.4.2.2. Controls
- 1.5. PROCEDURE
- 1.5.1. Number of animals
- 1.5.2. Treatment schedule
- 1.5.3. Dose levels
- 1.5.4. Limit test
- 1.5.5. Administration of doses
- 1.5.6. Chromosome preparation
- 1.5.7. Analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.24. MOUSE SPOT TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Experimental animals
- Number and sex
- Use of negative and positive controls
- Route of administration
- Dose levels
- Procedure
- Analysis
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.25. MOUSE HERITABLE TRANSLOCATION
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- Preparations
- Route of administration
- Experimental animals
- Number of animals
- Use of negative and positive controls
- Dose levels
- Procedure
- Treatment and mating
- Testing for translocation heterozygosity
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.26. SUB-CHRONIC ORAL TOXICITY TESTREPEATED DOSE 90 – DAY ORAL TOXICITY STUDY IN RODENTS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations of animals
- 1.4.2. Preparations of doses
- 1.4.3. Test conditions
- 1.4.3.1. Experimental animals
- 1.4.3.2. Number and sex
- 1.4.3.3. Dose levels
- 1.4.3.4. Limit test
- 1.5. PROCEDURE
- 1.5.1. Administration of doses
- 1.5.2. Observations
- 1.5.2.1. Body weight and food/water consumption
- 1.5.2.2. Haematology and clinical biochemistry
- 1.5.2.3. Gross necropsy
- 1.5.2.4. Histopathology
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Test species:
- 2.2.3. Test conditions:
- 2.2.4. Results:
- 3. REFERENCES
- B.27. SUB-CHRONIC ORAL TOXICITY TEST REPEATED DOSE 90-DAY ORAL TOXICITY STUDY IN NON-RODENTS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Selection of animal species
- 1.4.2. Preparation of animals
- 1.4.3. Preparations of doses
- 1.5. PROCEDURE
- 1.5.1. Number and sex of animals
- 1.5.2. Dosage
- 1.5.3. Limit test
- 1.5.4. Administration of doses
- 1.5.5. Observations
- 1.5.5.1. Body weight and food/water consumption
- 1.5.5.2. Haematology and clinical biochemistry
- 1.5.5.3. Gross necropsy
- 1.5.5.4. Histopathology
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Test species:
- 2.2.3. Test conditions:
- 2.2.4. Results:
- B.28. SUB-CHRONIC DERMAL TOXICITY STUDY 90-DAY REPEATED DERMAL DOSE STUDY USING RODENT SPECIES
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2.
- Test conditions
- 1.6.2.1.
- Experimental animals
- 1.6.2.2.
- Number and sex
- 1.6.2.3.
- Dose levels
- 1.6.3.
- Limit test
- 1.6.4.
- Observation period
- 1.6.5.
- Procedure
- Gross necropsy
- Histopathological examination
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.29. SUB-CHRONIC INHALATION TOXICITY STUDY 90-DAY REPEATED INHALATION DOSE STUDY USING RODENT SPECIES
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Preparations
- 1.6.2.
- Test conditions
- 1.6.2.1.
- Experimental animals
- 1.6.2.2.
- Number and sex
- 1.6.2.3.
- Exposure concentrations
- 1.6.2.4.
- Exposure time
- 1.6.2.5.
- Equipment
- 1.6.2.6.
- Observation period
- 1.6.3.
- Procedure
- Gross necropsy
- Histopathological examination
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.30. CHRONIC TOXICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Preparations
- 1.6.2.
- Test conditions
- 1.6.2.1.
- Experimental animals
- 1.6.2.2.
- Number and sex
- 1.6.2.3.
- Dose levels and frequency of exposure
- 1.6.2.4.
- Controls
- 1.6.2.5.
- Route of administration
- 1.6.2.6.
- Oral studies
- 1.6.2.7.
- Inhalation studies
- 1.6.2.8.
- Exposure chambers
- 1.6.2.9.
- Duration of study
- 1.6.3.
- Procedure
- Observations
- Haematological examination
- Urinalysis
- Clinical chemistry
- Gross necropsy
- Histopathology
- 2. DATA
- 3. REPORTING
- TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.31. PRENATAL DEVELOPMENTAL TOXICITY STUDY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCE
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Selection of animal species
- 1.5.2. Housing and feeding conditions
- 1.5.3. Preparation of the animals
- 1.6. PROCEDURE
- 1.6.1. Number and sex of animals
- 1.6.2. Preparation of doses
- 1.6.3. Dosage
- 1.6.4. Limit test
- 1.6.5. Administration of doses
- 1.6.6. Observations of the dams
- 1.6.7. Body weight and food consumption
- 1.6.8. Post-mortem examination
- 1.6.9. Examination of uterine contents
- 1.6.10. Examination of foetuses
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION OF RESULTS
- 2.3. INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.32. CARCINOGENICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Experimental animals
- 1.6.2.
- Number and sex
- 1.6.3.
- Dose levels and frequency of exposure
- 1.6.4.
- Controls
- 1.6.5.
- Route of administration
- 1.6.5.1. Oral studies
- 1.6.5.2. Dermal studies
- 1.6.5.3. Inhalation studies
- 1.6.6.
- Exposure chambers
- 1.6.7.
- Duration of study
- 1.6.8.
- Procedure
- 1.6.8.1.
- Observations
- 1.6.8.2.
- Clinical examinations
- Haematology
- Gross necropsy
- Histopathology
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.33. COMBINED CHRONIC TOXICITY/CARCINOGENICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Experimental animals
- 1.6.2.
- Number and sex
- 1.6.3.
- Dose levels and frequency of exposure
- 1.6.4.
- Controls
- 1.6.5.
- Route of administration
- 1.6.5.1. Oral tests
- 1.6.5.2. Dermal tests
- 1.6.5.3. Inhalation tests
- 1.6.6.
- Exposure chambers
- 1.6.7.
- Duration of test
- 1.6.8.
- Procedure
- 1.6.8.1.
- Observations
- 1.6.8.2.
- Clinical examinations
- Haematology
- Urinalysis
- Clinical chemistry
- Gross necropsy
- Histopathology
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.34. ONE-GENERATION REPRODUCTION TOXICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Preparations
- 1.6.2.
- Experimental animals
- Selection of species
- Number and sex
- 1.6.3.
- Test conditions
- 1.6.3.1.
- Dose levels
- 1.6.3.2.
- Limit test
- 1.6.3.3.
- Performance of the test
- Experimental schedules
- Mating procedure
- Litter sizes
- 1.6.4.
- Observations
- 1.6.5.
- Pathology
- 1.6.5.1.
- Necropsy
- 1.6.5.2.
- Histopathology
- 2. DATA
- 3. REPORTING
- TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.35. TWO-GENERATION REPRODUCTION TOXICITY STUDY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. PRINCIPLE OF THE TEST METHOD
- 1.3. DESCRIPTION OF THE TEST METHOD
- 1.3.1. Selection of animal species
- 1.3.2. Housing and feeding conditions
- 1.3.3. Preparation of animals
- 1.4. PROCEDURE
- 1.4.1. Number and sex of animals
- 1.4.2. Preparation of doses
- 1.4.3. Dosage
- 1.4.4. Limit test
- 1.4.5. Administration of doses
- 1.4.6. Experimental schedules
- 1.4.7. Mating procedure
- 1.4.7.1. Parental (P) mating
- 1.4.7.2. F1 mating
- 1.4.7.3. Second mating
- 1.4.7.4. Litter size
- 1.5. OBSERVATIONS
- 1.5.1. Clinical observations
- 1.5.2. Body weight and food/water consumption of parent animals
- 1.5.3. Oestrus cycle
- 1.5.4. Sperm parameters
- 1.5.5. Offspring
- 1.5.6. Gross necropsy
- 1.5.7. Organ weights
- 1.5.8. Histopathology
- 1.5.8.1. Parental animals
- 1.5.8.2. Weanlings
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION OF RESULTS
- 2.3. INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.36. TOXICOKINETICS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Preparations
- 1.6.2. Test conditions
- 1.6.2.1. Experimental animals
- 1.6.2.2. Number and sex
- 1.6.3. Dose levels
- 1.6.4. Route of administration
- 1.6.5. Observation period
- 1.6.6. Procedure
- Absorption
- Distribution
- Excretion
- Metabolism
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. EVALUATION AND INTERPRETATION
- 4. REFERENCES
- B.37. DELAYED NEUROTOXICITY OF ORGANOPHOSPHORUS SUBSTANCES FOLLOWING ACUTE EXPOSURE
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Preparations
- 1.5.2. Test conditions
- 1.5.2.1. Test animals
- 1.5.2.2. Number and sex
- 1.5.2.3. Dose levels
- 1.5.2.4. Limit test
- 1.5.3. Observation period
- 1.5.4. Procedure
- General observation
- Body weight
- Biochemistry
- Gross necropsy
- Histopathological examination
- 2. DATA
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.38. DELAYED NEUROTOXICITY OF ORGANOPHOSPHORUS SUBSTANCES 28-DAY REPEATED DOSE STUDY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.2. Test conditions
- 1.4.2.1. Test animals
- 1.4.2.2. Number and sex
- 1.4.2.3. Dose levels
- 1.4.2.4. Limit test
- 1.4.2.5. Observation period
- 1.4.3. Procedure
- General observations
- Body weight
- Biochemistry
- Gross necropsy
- Histopathological examination
- 2. DATA
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.39. UNSCHEDULED DNA SYNTHESIS (UDS) TEST WITH MAMMALIAN LIVER CELLS IN VIVO
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE METHOD
- 1.4.1. Preparations
- 1.4.1.1. Selection of animal species
- 1.4.1.2. Housing and feeding conditions
- 1.4.1.3. Preparation of the animals
- 1.4.1.4. Test substance/Preparation
- 1.4.2. Test conditions
- 1.4.2.1. Solvent/Vehicle
- 1.4.2.2. Controls
- 1.5. PROCEDURE
- 1.5.1. Number and sex of animals
- 1.5.2. Treatment schedule
- 1.5.3. Dose levels
- 1.5.4. Limit test
- 1.5.5. Administration of doses
- 1.5.6. Preparation of liver cells
- 1.5.7. Determination of UDS
- 1.5.8. Analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- B.40.
- IN VITRO
- SKIN CORROSION: TRANSCUTANEOUS ELECTRICAL RESISTANCE TEST (TER)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Animals
- 1.5.2. Preparation of the skin discs
- 1.5.3. Application of the test and control substances
- 1.5.4. TER measurements
- 1.5.5. Dye binding methods
- 1.5.5.1. Sulforhodamine B dye application and removal
- 1.5.5.2. Calculation of dye content
- 2. DATA
- 2.1. INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Figure 1
- Apparatus for the rat skin TER assay
- Figure 2
- Dimensions of the polytetrafluoroethylene (PFTE) and receptor tubes and electrodes used
- B.40 BIS.
- IN VITRO
- SKIN CORROSION: HUMAN SKIN MODEL TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.4.1. Procedure
- 1.4.1.1. Human skin models
- 1.4.1.1.1.
- General model conditions:
- 1.4.1.1.2.
- Functional model conditions:
- 1.4.1.2. Application of the test and control substances
- 1.4.1.3. Cell viability measurements
- 2. DATA
- 2.1. INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.41.
- IN VITRO
- 3T3 NRU PHOTOTOXICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Preparations
- 1.4.1.1. Cells
- 1.4.1.2. Media and culture conditions
- 1.4.1.3. Preparation of cultures
- 1.4.1.4. Preparation of test substance
- 1.4.1.5. Irradiation conditions
- 1.4.1.5.1.
- Light source
- 1.4.1.5.2.
- Dosimetry
- 1.4.2. Test conditions
- 1.4.2.1. Test substance concentrations
- 1.4.2.2. Controls
- 1.4.2.2.1.
- Radiation sensitivity of the cells, establishing of historical data:
- 1.4.2.2.2.
- Radiation sensitivity, check of current test:
- 1.4.2.2.3.
- Viability of solvent controls:
- 1.4.2.2.4.
- Positive control:
- 1.4.3. Test procedure (6)(7)(8)(16)(17):
- 1.4.3.1. 1st day:
- 1.4.3.2. 2nd day:
- 1.4.3.3. 3rd day:
- 1.4.3.3.1.
- Microscopic evaluation
- 1.4.3.3.2.
- Neutral Red uptake test
- 2. DATA
- 2.1. QUALITY AND QUANTITY OF DATA
- 2.2. EVALUATION OF RESULTS
- 2.3. INTERPRETATION OF RESULTS
- 2.4. INTERPRETATION OF DATA
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- Appendix 1
- Role of the 3T3 NRU PT in a sequential approach to the phototoxicity testing of chemicals
- Appendix 2
- Figure 1
- Spectral power distribution of a filtered solar simulator
- Figure 2
- Irradiation sensivity of Balb/c 3T3 cells (as measured in the UVA range)
- B.42. SKIN SENSITISATION: LOCAL LYMPH NODE ASSAY
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. PRINCIPLE OF THE TEST METHOD
- 1.3. DESCRIPTION OF THE TEST METHOD
- 1.3.1. Preparations
- 1.3.1.1. Housing and feeding conditions
- 1.3.1.2. Preparation of animals
- 1.3.2. Test conditions
- 1.3.2.1. Experimental animals
- 1.3.2.2. Reliability check
- 1.3.2.3. Number of animals, dose levels and vehicle selection.
- 1.3.3. Test procedure
- 1.3.3.1. Experimental schedule
- 1.3.3.2. Preparation of cell suspensions
- 1.3.3.3. Determination of cell proliferation (incorporated radioactivity)
- 1.3.3.4. Observations
- 1.3.3.4.1.
- Clinical observations
- 1.3.3.4.2.
- Body weights
- 1.3.4. Calculation of results
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.43. NEUROTOXICITY STUDY IN RODENTS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Selection of animal species
- 1.4.2. Housing and feeding conditions
- 1.4.3. Preparation of animals
- 1.4.4. Route of administration and preparation of doses
- 1.5. PROCEDURES
- 1.5.1. Number and sex animals
- 1.5.2. Treatment and control group
- 1.5.3. Reliability check
- 1.5.4. Dose selection
- 1.5.5. Limit test
- 1.5.6. Administration of doses
- 1.6. OBSERVATION
- 1.6.1. Frequency of observations and tests
- 1.6.1.1. Observations of general health condition and mortality/morbidity
- 1.6.1.2. Detailed clinical observations
- 1.6.1.3. Functional tests
- 1.6.2. Body weight and food/water consumption
- 1.6.3. Ophthalmology
- 1.6.4. Haematology and clinical biochemistry
- 1.6.5. Histopathology
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- B.44. SKIN ABSORPTION:
- IN VIVO
- METHOD
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE METHOD
- 1.4.1. Selection of animal species
- 1.4.2. Number and sex of animals
- 1.4.3. Housing and feeding conditions
- 1.4.4. Preparation of animals
- 1.4.5. Test substance
- 1.4.6. Test preparation
- 1.4.7. Application to the skin
- 1.4.8. Duration of exposure and sampling
- 1.4.9. Terminal procedures
- 1.4.10. Analysis
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Figure 1
- An example of a design of a typical device used to define and protect dermal application site during
- in vivo
- percutaneous absorption studies
- B.45. SKIN ABSORPTION:
- IN VITRO
- METHOD
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. DESCRIPTION OF THE TEST METHOD
- 1.4.1. Diffusion cell
- 1.4.2. Receptor fluid
- 1.4.3. Skin preparations
- 1.4.4. Skin preparation integrity
- 1.4.5. Test substance
- 1.4.6. Test preparation
- 1.4.7. Test substances concentrations and formulations
- 1.4.8. Application to the skin
- 1.4.9. Temperature
- 1.4.10. Duration of exposure and sampling
- 1.4.11. Terminal procedures
- 1.4.12. Analysis
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Figure 1
- An example of a typical design of a static diffusion cell for
- in vitro
- percutaneous absorption studies
- PART C: METHODS FOR THE DETERMINATION OF ECOTOXICITY
- C.1. ACUTE TOXICITY FOR FISH
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Reagents
- 1.6.1.1. Solutions of test substances
- 1.6.1.2. Holding and dilution water
- 1.6.2. Apparatus
- 1.6.3. Test fish
- 1.6.3.1. Holding
- 1.6.3.2. Mortality
- 1.6.4. Adaptation
- 1.6.5. Test procedure
- Limit test
- 2. DATA AND EVALUATION
- 3. REPORTING
- 4. REFERENCES
- Appendix 1
- Reconstituted water
- Example of a suitable dilution water
- Stock solutions
- Reconstituted dilution water
- Appendix 2
- Fish species recommended for testing
- Collection
- Appendix 3
- Example of concentration: percentage mortality
- C.2.
- DAPHNIA
- SP. ACUTE IMMOBILISATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. REFERENCE SUBSTANCES
- 1.6. QUALITY CRITERIA
- 1.7. DESCRIPTION OF THE TEST METHOD
- 1.7.1. Apparatus
- 1.7.2. Test organism
- 1.7.3. Holding and dilution water
- 1.7.4. Test solutions
- 1.8. PROCEDURE
- 1.8.1. Conditions of exposure
- 1.8.1.1. Test groups and controls
- 1.8.1.2. Test concentrations
- 1.8.1.3. Incubation conditions
- 1.8.1.4. Duration
- 1.8.2. Observations
- 1.8.3. Analytical measurements
- 1.9. LIMIT TEST
- 2. DATA
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Appendix 1
- SOME CHEMICAL CHARACTERISTICS OF AN ACCEPTABLE DILUTION WATER
- Appendix 2
- EXAMPLES OF SUITABLE RECONSTITUTED TEST WATER
- ISO Test water (1)
- Elendt M7 and M4 medium
- Acclimation to Elendt M4 and M7 medium
- Preparation
- Trace element
- M4 and M7 media
- C.3. ALGAL INHIBITION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST PROCEDURE
- 1.6.1. Reagents
- 1.6.1.1. Solutions of test substances
- 1.6.1.2. Test medium
- 1.6.2. Apparatus
- 1.6.3. Test organisms
- 1.6.4. Test procedure
- Initial cell density
- Concentrations of test substance
- Replicates and controls
- Performance of the test
- Testing volatile substances
- Limit test
- 2. DATA AND EVALUATION
- 2.1. COMPARISON OF AREAS UNDER THE GROWTH CURVES
- 2.2. COMPARISON OF GROWTH RATES
- 2.3. CALCULATION OF THE NOEC
- 3. REPORTING
- 4. REFERENCES
- Appendix 1
- Example of a procedure for the culturing of algae
- General observations
- Procedures for obtaining algal cultures
- Preparation of nutrient solutions (media):
- Stock culture:
- Pre-culture:
- Appendix 2
- C.4. DETERMINATION OF ‘READY’ BIODEGRADABILITY
- PART I. GENERAL CONSIDERATIONS
- I.1. INTRODUCTION
- I.2. SELECTION OF THE APPROPRIATE METHOD
- I.3. REFERENCE SUBSTANCES
- I.4. PRINCIPLE OF THE TEST METHODS
- I.5. QUALITY CRITERIA
- I.5.1. Reproducibility
- I.5.2. Validity of the test
- I.6. GENERAL PROCEDURES AND PREPARATIONS
- I.6.1. Dilution water
- I.6.2. Stock solutions of mineral components
- Stock solutions:
- I.6.3. Stock solutions of chemicals
- I.6.4. Inocula
- I.6.4.1. Inoculum from activated sludges
- I.6.4.2. Other sources of inoculum
- I.6.5. Pre-conditioning of inocula
- I.6.6. Abiotic controls
- I.6.7. Number of flasks
- I.7. DATA AND EVALUATION
- I.7.1. Degradation measured by means of DOC determination
- I.7.2. Degradation measured by means of specific analysis
- 1.7.3. Abiotic degradation
- I.8. REPORTING
- PART II. DOC DIE-AWAY TEST
- (Method C.4-A)
- II.1. PRINCIPLE OF THE METHOD
- II.2. DESCRIPTION OF THE METHOD
- II.2.1. Apparatus
- II.2.2. Preparation of mineral medium
- II.2.3. Preparation and pre-conditioning of inoculum
- II.2.4. Preparation of flasks
- II.2.5. Number of flasks in typical run
- II.2.6. Performance of the test
- II.3. DATA AND REPORTING
- II.3.1. Treatment of results
- II.3.2. Validity of results
- II.3.3. Reporting
- II.4. DATA SHEET
- DOC DIE-AWAY TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 5. CARBON DETERMINATIONS
- 6. EVALUATION OF RAW DATA
- 7. ABIOTIC CONTROL
- (optional)
- 8. SPECIFIC CHEMICAL ANALYSIS
- (optional)
- PART III. MODIFIED OECD SCREENING TEST
- (Method C.4-B)
- III.1. PRINCIPLE OF THE METHOD
- III.2. DESCRIPTION OF THE METHOD
- III.2.1.
- Apparatus
- III.2.2. Preparation of mineral medium
- III.2.3. Preparation and pre-conditioning of inoculum
- III.2.4. Preparation of flasks
- III.2.5. Number of flasks in typical run
- III.2.6. Performance of the test
- III.3. DATA AND REPORTING
- III.3.1. Treatment of results
- III.3.2.
- Validity of results
- III.3.3.
- Reporting
- III.4. DATA SHEET
- MODIFIED OECD SCREENING TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 5.
- CARBON DETERMINATIONS
- 6. EVALUATION OF RAW DATA
- 7. ABIOTIC CONTROL
- (optional)
- 8. SPECIFIC CHEMICAL ANALYSIS
- (optional)
- PART IV.
- CO
- 2
- EVOLUTION TEST (Method C.4-C)
- IV.1. PRINCIPLE OF THE METHOD
- IV.2. DESCRIPTION OF THE METHOD
- IV.2.1. Apparatus
- IV.2.2. Preparation of mineral medium
- IV.2.3. Preparation and pre-conditioning of inoculum
- IV.2.4. Preparation of flasks
- IV.2.5. Number of flasks in a typical run
- IV.2.6. Performance of the test
- IV.3. DATA AND REPORTING
- IV.3.1. Treatment of results
- IV.3.2. Validity of results
- IV.3.3. Reporting
- IV.4. DATA SHEET
- CARBON DIOXIDE EVOLUTION TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 6. CARBON ANALYSIS
- (optional)
- 7. ABIOTIC DEGRADATION
- (optional)
- PART V. MANOMETRIC RESPIROMETRY TEST
- (Method C.4-D)
- V.1. PRINCIPLE OF THE METHOD
- V.2. DESCRIPTION OF THE METHOD
- V.2.1. Apparatus
- V.2.2. Preparation of mineral medium
- V.2.3. Preparation and pre-conditioning of inoculum
- V.2.4. Preparation of flasks
- V.2.5. Number of flasks in a typical run
- V.2.6. Performance of the test
- V.3. DATA AND REPORTING
- V.3.1. Treatment of results
- V.3.2. Validity of results
- V.3.3. Reporting
- V.4.
- DATA SHEET
- MANOMETRIC RESPIROMETRY TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 5. OXYGEN UPTAKE: BIODEGRADABILITY
- 6. CORRECTION FOR NITRIFICATION
- (see Annex V)
- 7. CARBON ANALYSIS
- (optional)
- 8. SPECIFIC CHEMICAL
- (optional)
- 9. ABIOTIC DEGRADATION
- (optional)
- PART VI. CLOSED BOTTLE TEST
- (Method C.4-E)
- VI.1 PRINCIPLE OF THE TEST METHOD
- VI.2 DESCRIPTION OF THE METHOD
- VI.2.1. Apparatus
- VI.2.2. Preparation of mineral medium
- VI.2.3. Preparation of the inoculum
- VI.2.4. Preparation of flasks
- VI.2.5. Number of flasks in a typical run
- VI.2.6.
- Performance of the test
- VI.3. DATA AND REPORTING
- VI.3.1. Treatment of results
- VI.3.2. Validity of results
- VI.3.3. Reporting
- VI.4. DATA SHEET
- CLOSED BOTTLE TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 5. DO DETERMINATION
- 6. CORRECTION FOR NITRIFICATION
- (see Annex V)
- 7. DO DEPLETION: % DEGRADATION
- 8. BLANK DO DEPLETIONS
- PART VII. M.I.T.I. TEST
- (Method C.4-F)
- VII.1. PRINCIPLE OF THE METHOD
- VII.2. DESCRIPTION OF THE METHOD
- VII.2.1. Apparatus
- VII.2.2. Preparation of mineral medium
- VII.2.3. Preparation of inoculum
- VII.2.4. Preparation of flasks
- VII.2.5. Performance of the test
- VII.3. DATA AND REPORTING
- VII.3.1. Treatment of results
- VII.3.2. Validity of results
- VII.3.3. Reporting
- VII.4. DATA SHEET
- MITI (I) TEST
- 3. TEST SUBSTANCE
- 4. INOCULUM
- 5. OXYGEN UPTAKE: BIODEGRADABILITY
- 6. CARBON ANALYSIS
- (optional)
- 7. SPECIFIC CHEMICAL ANALYTICAL DATA
- 8. REMARKS
- Appendix 1
- ABBREVIATIONS AND DEFINITIONS
- Primary biodegradation:
- Ultimate biodegradation (aerobic):
- Readily biodegradable:
- Inherently biodegradable:
- Treatability:
- Lag time
- Degradation time
- 10-day window
- Appendix 2
- CALCULATION AND DETERMINATION OF SUITABLE SUMMARY PARAMETERS
- 1. Carbon content
- 2. Theoretical oxygen demand (ThOD)
- 3. Chemical Oxygen Demand (COD)
- 4. Dissolved organic carbon (DOC)
- Remarks:
- BIBLIOGRAPHY
- Appendix 3
- EVALUATION OF THE BIODEGRADABILITY OF POORLY SOLUBLE SUBSTANCES
- BIBLIOGRAPHY
- Appendix 4
- EVALUATION OF THE BIODEGRADABILITY OF CHEMICALS SUSPECTED TO BE TOXIC TO THE INOCULUM
- BIBLIOGRAPHY
- Appendix 5
- CORRECTION FOR OXYGEN UPTAKE FOR INTERFERENCE BY NITRIFICATION
- C.5 DEGRADATION — BIOCHEMICAL OXYGEN DEMAND
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 2. DATA AND EVALUATION
- 3. REPORTING
- 4.
- REFERENCES
- C.6. DEGRADATION — CHEMICAL OXYGEN DEMAND
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 2. DATA AND EVALUATION
- 3. REPORTING
- 4. REFERENCES
- C.7. DEGRADATION — ABIOTIC DEGRADATION: HYDROLYSIS AS A FUNCTION OF PH
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. APPLICABILITY OF THE METHOD
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. INFORMATION ON THE TEST SUBSTANCE
- 1.6. REFERENCE SUBSTANCES
- 1.7. QUALITY CRITERIA
- 1.7.1. Recovery
- 1.7.2. Repeatability and sensitivity of analytical method
- 1.7.3. Confidence intervals for hydrolysis kinetic data
- 1.8. DESCRIPTION OF THE TEST METHOD
- 1.8.1. Equipment and apparatus
- 1.8.2. Test substance application
- 1.8.3. Buffer solutions
- 1.8.4. Test conditions
- 1.8.4.1. Test temperature
- 1.8.4.2. Light and oxygen
- 1.8.4.3. Test duration
- 1.8.5. Performance of the test
- 1.8.5.1. Preliminary test (Tier 1)
- 1.8.5.2. Hydrolysis of unstable substances (Tier 2)
- 1.8.5.3. Identification of hydrolysis products (Tier 3)
- 1.8.5.4. Optional tests
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Appendix 1
- Tiered hydrolysis test scheme
- Appendix 2
- Definitions and units
- Appendix 3
- Buffer Systems
- A. CLARK AND LUBS:
- Buffer mixtures of CLARK and LUBS
- (14)
- Buffer mixtures of CLARK and LUBS (Continued)
- B. KOLTHOFF AND VLEESCHHOUWER:
- Citrate buffers of KOLTHOFF and VLEESCHHOUWER
- C. SÖRENSEN:
- Borate mixtures of SÖRENSEN
- Phosphate mixtures of SÖRENSEN
- C.8. TOXICITY FOR EARTHWORMS
- ARTIFICIAL SOIL TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITION AND UNIT
- 1.3. REFERENCE SUBSTANCE
- 1.4. PRINCIPLE OF THE TEST
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Materials
- 1.6.1.1.
- Test substrate
- 1.6.1.2.
- Test containers
- 1.6.2.
- Test conditions
- 1.6.3.
- Test procedure
- Test concentrations
- Range finding test
- Definitive test
- Mixture of the basic test substrate and the test substance
- 1.6.4.
- Test organisms
- 2. DATA
- 2.1. TREATMENT AND EVALUATION OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Appendix
- Breeding and keeping of the worms before testing
- Keeping and breeding conditions
- C.9. BIODEGRADATION
- ZAHN-WELLENS TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST PROCEDURE
- 1.6.1.
- Preparations
- 1.6.1.1.
- Reagents
- 1.6.1.2.
- Apparatus
- 1.6.1.3.
- Preparation of the inoculum
- 1.6.1.4.
- Preparation of the test solutions
- 1.6.2.
- Performance of the test
- Functional control of activated sludge
- Adaptation
- Analytical means
- 2. DATA AND EVALUATION
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF RESULTS
- 4. REFERENCES
- Appendix
- EVALUATION EXAMPLE
- Figure 1
- Examples of biodegradation curves
- Figure 2
- Examples of sludge adaptation
- C.10. BIODEGRADATION
- ACTIVATED SLUDGE SIMULATION TESTS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.1.1.
- General remarks
- 1.1.2.
- Determination of ultimate biodegradability (DOC/COD analysis)
- 1.1.3.
- Determination of primary biodegradability (specific analysis)
- 1.2. DEFINITIONS AND UNITS
- 1.2.1.
- DOC/COD analysis
- 1.2.2.
- Specific analysis
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHODS
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Preparation
- 1.6.1.1.
- Apparatus
- 1.6.1.2.
- Filtration
- 1.6.1.3.
- Sewage
- 1.6.1.4.
- Stock solution of test material
- 1.6.1.5.
- Inoculum
- 1.6.2.
- Procedure
- 1.6.2.1.
- Running-in period: Sludge formation/stabilisation of the units
- 1.6.2.2.
- Test procedure
- 1.6.2.3.
- Analysis
- 2. DATA AND EVALUATION
- 2.1. COUPLED UNITS MODE
- 2.2. NON-COUPLED UNITS MODE
- 2.2.1.
- Using COD/DOC determinations
- 2.2.2.
- Using specific analysis
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF RESULTS
- 4. REFERENCES
- Appendix 1
- Figure 1
- Figure 2
- Appendix 2
- Figure 1
- Equipment used for assessing biodegradability
- Figure 2
- Details of three-litre porous-pot aeration vessel
- Appendix 3
- Operating conditions for the Activated Sludge Simulation Test
- Check in each group
- Apparatus
- Mode of operation
- Transinoculation
- Mean retention time
- Base nutrient
- Inoculum
- Test material addition
- Analysis
- C.11. BIODEGRADATION
- ACTIVATED SLUDGE RESPIRATION INHIBITION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- Recommendation
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1.
- Reagents
- 1.6.1.1.
- Solutions of the test substance
- 1.6.1.2.
- Solution of control substance
- 1.6.1.3.
- Synthetic sewage
- 1.6.2.
- Apparatus
- 1.6.3.
- Preparation of the inoculum
- 1.6.4.
- Performance of the test
- 2. DATA AND EVALUATION
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF DATA
- 4. REFERENCES
- C.12. BIODEGRADATION
- MODIFIED SCAS TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. QUALITY CRITERIA
- 1.6. DESCRIPTION OF THE TEST PROCEDURE
- 1.6.1.
- Preparations
- 1.6.2.
- Test conditions
- 1.6.3.
- Performance of the test
- 2. DATA AND EVALUATION
- 3. REPORTING
- 3.1. TEST REPORT
- 3.2. INTERPRETATION OF RESULTS
- 4. REFERENCES
- Appendix 1
- SCAS test: example of results
- Appendix 2
- Example of test apparatus
- Figure 1
- C.13 BIOCONCENTRATION: FLOW-THROUGH FISH TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. VALIDITY OF THE TEST
- 1.6. REFERENCE COMPOUNDS
- 1.7. DESCRIPTION OF THE TEST METHOD
- 1.7.1.
- Apparatus
- 1.7.2.
- Water
- 1.7.3.
- Test Solutions
- 1.7.4.
- Selection of species
- 1.7.5.
- Holding of fish
- 1.8. PERFORMANCE OF THE TEST
- 1.8.1.
- Preliminary test
- 1.8.2.
- Conditions of exposure
- 1.8.2.1.
- Duration of uptake phase
- 1.8.2.2.
- Duration of the depuration phase
- 1.8.2.3.
- Numbers of test fish
- 1.8.2.4.
- Loading
- 1.8.2.5.
- Feeding
- 1.8.2.6.
- Light and temperature
- 1.8.2.7.
- Test concentrations
- 1.8.2.8.
- Controls
- 1.8.3.
- Frequency of water quality measurements
- 1.8.4.
- Sampling and analysis of fish and water
- 1.8.4.1.
- Fish and water sampling schedule
- 1.8.4.2.
- Sampling and Sample Preparation
- 1.8.4.3.
- Quality of Analytical method
- 1.8.4.4.
- Analysis of Fish Sample
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. INTERPRETATION OF RESULTS
- 3. REPORTING
- 3.1. TEST SUBSTANCE:
- 3.2. TEST SPECIES
- 3.3. TEST CONDITIONS:
- 3.4. RESULTS:
- 4. REFERENCES
- Appendix 1
- Chemical characteristics of an acceptable dilution water
- Appendix 2
- Fish species recommended for testing
- Collection
- Appendix 3
- Prediction of the duration of the uptake and depuration phases
- 1. Prediction of the duration of the uptake phase
- 2. Prediction of the duration of the depuration phase
- Literature
- (of Appendix 3)
- Appendix 4
- Theoretical example of sampling schedule for bioconcentration tests of substances with log P
- ow
- = 4
- Appendix 5
- Model discrimination
- Graphical method for determination of depuration (loss) rate constant k
- 2
- Graphical method for determination of uptake rate constant k
- 1
- Computer method for calculation of uptake and depuration (loss) rate constants
- C.14. FISH JUVENILE GROWTH TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. VALIDITY OF THE TEST
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Apparatus
- 1.6.2. Water
- 1.6.3. Test solutions
- 1.6.4. Selection of species
- 1.6.5. Holding of fish
- 1.7. TEST DESIGN
- 1.7.1. Design for analysis by regression
- 1.7.2. Design for estimation of a NOEC/LOEC using Analysis of Variance (ANOVA)
- 1.8. PROCEDURE
- 1.8.1. Selection and weighing of test fish
- 1.8.2. Conditions of exposure
- 1.8.2.1. Duration
- 1.8.2.2. Loading rates and stocking densities
- 1.8.2.3. Feeding
- 1.8.2.4. Light and temperature
- 1.8.3. Test concentrations
- 1.8.4. Controls
- 1.8.5. Frequency of analytical determinations and measurements
- 1.8.6. Observations
- 2. DATA AND REPORTING
- 2.1. TREATMENT OF RESULTS
- 2.1.1. Analysis of results by regression (concentration-response modelling)
- 2.1.2. Analysis of results for the estimation of the LOEC
- 2.2. INTERPRETATION OF RESULTS
- 2.3. TEST REPORT
- 2.3.1. Test substance:
- 2.3.2. Test species:
- 2.3.3. Test conditions:
- 2.3.4. Results:
- 3.
- REFERENCES
- Appendix 1
- FISH SPECIES RECOMMENDED FOR TESTING AND SUITABLE TEST CONDITIONS
- Appendix 2
- SOME CHEMICAL CHARACTERISTICS OF AN ACCEPTABLE DILUTION WATER
- Appendix 3
- Logarithmic series of concentrations suitable for toxicity test (9)
- C.15. FISH, SHORT-TERM TOXICITY TEST ON
- EMBRYO
- AND SAC-FRY STAGES
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. VALIDITY OF THE TEST
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Test chambers
- 1.6.2. Selection of fish species
- 1.6.3. Holding of the brood fish
- 1.6.4. Handling of embryos and larvae
- 1.6.5. Water
- 1.6.6. Test solutions
- 1.7. PROCEDURE
- 1.7.1. Conditions of exposure
- 1.7.1.1. Duration
- 1.7.1.2. Loading
- 1.7.1.3. Light and temperature
- 1.7.2. Test concentrations
- 1.7.3. Controls
- 1.7.4. Frequency of analytical determinations and measurements
- 1.7.5. Observations
- 1.7.5.1. Stage of embryonic development
- 1.7.5.2. Hatching and survival
- 1.7.5.3. Abnormal appearance
- 1.7.5.4. Abnormal behaviour
- 1.7.5.5. Length
- 1.7.5.6. Weight
- 2. DATA AND REPORTING
- 2.1. TREATMENT OF RESULTS
- 2.2. INTERPRETATION OF RESULTS
- 2.3. THE TEST REPORT
- 2.3.1. Test substance:
- 2.3.2. Test species:
- 2.3.3. Test conditions:
- 2.3.4. Results:
- 3. REFERENCES
- Appendix 1
- GUIDANCE ON PERFORMANCE OF A TOXICITY TEST ON EMBRYOS AND SAC-FRY OF ZEBRAFISH
- (BRACHYDANIO RERIO)
- INTRODUCTION
- CONDITIONS OF PARENTAL FISH, REPRODUCTION AND EARLY-LIFE STAGES
- CALCULATIONS AND STATISTICS
- LC
- 50
- AND EC
- 50
- DETERMINATIONS
- ESTIMATION OF LOEC AND NOEC
- REFERENCES
- Appendix 2
- TEST CONDITIONS, DURATION AND SURVIVAL CRITERIA FOR RECOMMENDED SPECIES
- Appendix 3
- Test conditions, duration and survival criteria for other well documented species
- Appendix 4
- SOME CHEMICAL CHARACTERISTICS OF AN ACCEPTABLE DILUTION WATER
- C.16. HONEYBEES — ACUTE ORAL TOXICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. VALIDITY OF THE TEST
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Collection of bees
- 1.5.2. Housing and feeding conditions
- 1.5.3. Preparation of bees
- 1.5.4. Preparation of doses
- 1.6. PROCEDURE
- 1.6.1. Test and control groups
- 1.6.2. Toxic standard
- 1.6.3. Exposure
- 1.6.3.1. Administration of doses
- 1.6.3.2. Duration
- 1.6.4. Observations
- 1.6.5. Limit test
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Test species:
- 2.2.3. Test conditions:
- 2.2.4. Results:
- 3. REFERENCES
- C.17. HONEYBEES — ACUTE CONTACT TOXICITY TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. VALIDITY OF THE TEST
- 1.5. DESCRIPTION OF THE TEST METHOD
- 1.5.1. Collection of bees
- 1.5.2. Housing and feeding conditions
- 1.5.3. Preparation of bees
- 1.5.4. Preparation of doses
- 1.6. PROCEDURE
- 1.6.1. Test and control groups
- 1.6.2. Toxic standard
- 1.6.3. Exposure
- 1.6.3.1. Administration of doses
- 1.6.3.2. Duration
- 1.6.4. Observations
- 1.6.5. Limit test
- 2. DATA AND REPORTING
- 2.1. DATA
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Test species:
- 2.2.3. Test conditions:
- 2.2.4. Results:
- 3. REFERENCES
- C.18. ADSORPTION/DESORPTION USING A BATCH EQUILIBRIUM METHOD
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. SCOPE
- 1.3. DEFINITIONS AND UNITS
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. INFORMATION ON THE TEST SUBSTANCE
- 1.6. APPLICABILITY OF THE TEST
- 1.7. DESCRIPTION OF THE METHOD
- 1.7.1. Apparatus and chemical reagents
- 1.7.2. Characterisation and selection of soils
- 1.7.3. Collection and storage of soil samples
- 1.7.3.1. Collection
- 1.7.3.2. Storage
- 1.7.3.3. Handling and preparation of soil samples for the test
- 1.7.4. Preparation of the test substance for application to soil
- 1.8. PREREQUISITES FOR PERFORMING THE ADSORPTION/DESORPTION TEST
- 1.8.1. The analytical method
- 1.8.2. The selection of optimal soil/solution ratios
- 1.9. PERFORMANCE OF THE TEST
- 1.9.1. Test conditions
- 1.9.2. Tier 1 — Preliminary study
- 1.9.2.1. Selection of optimal soil/solution ratios
- 1.9.2.2. Determination of adsorption equilibration time and of the amount of test substance adsorbed at equilibrium
- 1.9.2.3. Adsorption on the surface of the test vessel and stability of the test substance
- 1.9.3. Tier 2 — Adsorption kinetics at one concentration of the test substance
- 1.9.4. Tier 3 — Adsorption isotherms and desorption kinetics/desorption isotherms
- 1.9.4.1.
- Adsorption isotherms
- 1.9.4.2.
- Desorption kinetics
- 1.9.4.3.
- Desorption isotherms
- 2. DATA AND REPORTING
- 2.1. ADSORPTION
- 2.1.1. Adsorption isotherms
- 2.1.2. Mass balance
- 2.2. DESORPTION
- 2.2.1. Desorption isotherms
- 2.2.2. Test report
- 3. REFERENCES
- Appendix 1
- Testing scheme
- Appendix 2
- INFLUENCE OF ACCURACY OF ANALYTICAL METHOD AND CONCENTRATION CHANGE ON ACCURACY OF ADSORPTION RESULTS
- Appendix 3
- ESTIMATION TECHNIQUES FOR K
- D
- Appendix 4
- CALCULATIONS FOR DEFINING THE CENTRIFUGATION CONDITIONS
- Fig. 1a.
- Fig. 1b.
- Appendix 5
- CALCULATION OF ADSORPTION A (%) AND DESORPTION D (%)
- ADSORPTION A (A%)
- a) Parallel method
- b) Serial method
- DESORPTION D (A %)
- (a) Parallel method
- (b) Serial method
- Appendix 6
- ADSORPTION-DESORPTION IN SOILS: DATA REPORTING SHEETS
- Suitability of the analytical method
- Adsorption test: test samples
- Adsorption test: blanks and control
- Mass balance
- Adsorption isotherms
- Desorption test
- C.19. ESTIMATION OF THE ADSORPTION COEFFICIENT (
- K
- OC
- ) ON SOIL AND ON SEWAGE SLUDGE USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. APPLICABILITY OF THE TEST
- 1.6. QUALITY CRITERIA
- 1.6.1. Accuracy
- 1.6.2. Repeatability
- 1.6.3. Reproducibility
- 1.7. DESCRIPTION OF THE TEST METHOD
- 1.7.1. Preliminary Estimation of the Adsorption Coefficient
- 1.7.2. Apparatus
- 1.7.3. Mobile phases
- 1.7.4. Solutes
- 1.8. PERFORMANCE OF THE TEST
- 1.8.1. Test condition
- 1.8.2. Determination of dead time to
- 1.8.2.1. Determination of the dead time to by means of a homologous series
- 1.8.2.2. Determination of the dead time to by inert substances which are not retained by the column
- 1.8.3. Determination of the retention times tR
- 1.8.4. Evaluation
- 2. DATA AND REPORTING
- 3. REFERENCES
- Appendix
- C.20
- DAPHNIA MAGNA
- REPRODUCTION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS AND UNITS
- 1.3. PRINCIPLE OF THE TEST METHOD
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. VALIDITY OF THE TEST
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Apparatus
- 1.6.2. Test organism
- 1.6.3. Test medium
- 1.6.4. Test solutions
- 1.7. TEST DESIGN
- 1.8. PROCEDURE
- 1.8.1. Conditions of exposure
- 1.8.1.1. Duration
- 1.8.1.2. Loading
- 1.8.1.3. Number of animals
- 1.8.1.4. Feeding
- 1.8.1.5. Light
- 1.8.1.6. Temperature
- 1.8.1.7. Aeration
- 1.8.2. Test concentration
- 1.8.3. Controls
- 1.8.4. Test medium renewal
- 1.8.5. Observations
- 1.8.6. Offspring
- 1.8.7. Mortality
- 1.8.8. Other parameters
- 1.8.9. Frequency of analytical determinations and measurements
- 2. DATA AND REPORTING
- 2.1. TREATMENT OF RESULTS
- 2.2. TEST REPORT
- 2.2.1. Test substance:
- 2.2.2. Test species:
- 2.2.3. Test conditions:
- 2.2.4. Results:
- 3. REFERENCES
- Appendix 1
- PREPARATION OF FULLY DEFINED ELENDT M7 AND M4 MEDIA
- Acclimation to Elendt M7 and M4 media
- PREPARATION
- Trace elements
- M4 and M7 media
- Appendix 2
- TOTAL ORGANIC CARBON (TOC) ANALYSIS AND PRODUCTION OF A NOMOGRAPH FOR TOC CONTENT OF ALGAL FEED
- Appendix 3
- EXAMPLE DATA SHEET RECORDING MEDIUM RENEWAL, PHISICAL/CHEMICAL MONITORING DATA, FEEDING,
- DAPHNIA
- REPRODUCTION AND ADULT MORTALITY
- Appendix 4
- EXAMPLE DATA SHEET FOR RECORDING RESULTS OF CHEMICAL ANALYSIS
- (a) Measured concentrations
- (b) Measured concentrations as a percentage of nominal
- Appendix 5
- CALCULATION OF A TIME-WEIGHTED MEAN
- Time-weighted mean
- Figure 1: Example of time-weighted mean
- C.21. SOIL MICROORGANISMS: NITROGEN TRANSFORMATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. VALIDITY OF THE TEST
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Apparatus
- 1.6.2. Selection and number of soils
- 1.6.3. Collection and storage of soil samples
- 1.6.3.1. Collection
- 1.6.3.2. Storage
- 1.6.4. Handling and preparation of soil for the test
- 1.6.4.1. Pre-incubation
- 1.6.4.2. Physical-chemical characteristics
- 1.6.4.3. Amendment with organic substrate
- 1.6.5. Preparation of the test substance for the application to soil
- 1.6.6. Test concentrations
- 1.7. PERFORMANCE OF THE TEST
- 1.7.1. Conditions of exposure
- 1.7.1.1. Treatment and control
- 1.7.1.2. Incubation of soil samples
- 1.7.1.3. Test conditions and duration
- 1.7.2. Sampling and analysis of soils
- 1.7.2.1. Soil sampling schedule
- 1.7.2.2. Analysis of soil samples
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. INTERPRETATION OF RESULTS
- 3. REPORTING
- 4. REFERENCES
- C.22. SOIL MICROORGANISMS: CARBON TRANSFORMATION TEST
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. PRINCIPLE OF THE TEST METHOD
- 1.5. VALIDITY OF THE TEST
- 1.6. DESCRIPTION OF THE TEST METHOD
- 1.6.1. Apparatus
- 1.6.2. Selection and number of soils
- 1.6.3. Collection and storage of soil samples
- 1.6.3.1. Collection
- 1.6.3.2. Storage
- 1.6.4. Handling and preparation of soil for the test
- 1.6.4.1. Pre-incubation
- 1.6.4.2. Physical-chemical characteristics
- 1.6.5. Preparation of the test substance for the application to soil
- 1.6.6. Test concentrations
- 1.7. PERFORMANCE OF THE TEST
- 1.7.1. Conditions of exposure
- 1.7.1.1. Treatment and control
- 1.7.1.2. Incubation of soil samples
- 1.7.1.3. Test conditions and duration
- 1.7.2. Sampling and analysis of soils
- 1.7.2.1. Soil sampling schedule
- 1.7.2.2. Measurement of glucose-induced respiration rates
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- C.23. AEROBIC AND ANAEROBIC TRANSFORMATION IN SOIL
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. APPLICABILITY OF THE TEST
- 1.5. INFORMATION ON THE SUBSTANCE
- 1.6. PRINCIPLE OF THE TEST METHOD
- 1.7. QUALITY CRITERIA
- 1.7.1. Recovery
- 1.7.2. Repeatability and sensitivity of analytical method
- 1.7.3. Accuracy of transformation data
- 1.8. DESCRIPTION OF THE METHOD
- 1.8.1. Equipment and chemical reagents
- 1.8.2. Test substance application
- 1.8.3. Soils
- 1.8.3.1. Soil selection
- 1.8.3.2. Collection, handling, and storage of soils
- 1.9. PERFORMANCE OF THE TEST
- 1.9.1. Test conditions
- 1.9.1.1. Test temperature
- 1.9.1.2. Moisture content
- 1.9.1.3. Aerobic incubation conditions
- 1.9.1.4. Sterile aerobic conditions
- 1.9.1.5. Anaerobic incubation conditions
- 1.9.1.6. Paddy incubation conditions
- 1.9.1.7. Test duration
- 1.9.2. Performance of the test
- 1.9.3. Sampling and measurement
- 1.9.4. Optional tests
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 2.2. EVALUATION AND INTERPRETATION OF RESULTS
- 3. REPORTING
- TEST REPORT
- 4. REFERENCES
- Appendix 1
- WATER TENSION, FIELD CAPACITY (FC) AND WATER HOLDING CAPACITY (WHC)(47)
- Appendix 2
- SOIL MOISTURE CONTENTS (g water per 100 g dry soil) OF VARIOUS SOIL TYPES FROM VARIOUS COUNTRIES
- Appendix 3
- Figure 1
- Example of a flow-through apparatus to study transformation of chemicals in soil
- (53) (54)
- Figure 2
- Example of a biometer-type flask for studying the transformation of chemicals in soil
- (55)
- C.24. AEROBIC AND ANAEROBIC TRANSFORMATION IN AQUATIC SEDIMENT SYSTEMS
- 1. METHOD
- 1.1. INTRODUCTION
- 1.2. DEFINITIONS
- 1.3. REFERENCE SUBSTANCES
- 1.4. INFORMATION ON THE TEST SUBSTANCE
- 1.5. PRINCIPLE OF THE TEST METHOD
- 1.6. APPLICABILITY OF THE TEST
- 1.7. QUALITY CRITERIA
- 1.7.1. Recovery
- 1.7.2. Repeatability and sensitivity of analytical method
- 1.7.3. Accuracy of transformation data
- 1.8. DESCRIPTION OF THE METHOD
- 1.8.1. Test system and apparatus
- 1.8.2. Selection and number of aquatic sediments
- 1.8.2.1. Sediment selection
- 1.8.2.2. Characterisation of water-sediment samples
- 1.8.3. Collection, handling and storage
- 1.8.3.1. Collection
- 1.8.3.2. Handling
- 1.8.3.3. Storage
- 1.8.4. Preparation of the sediment/water samples for the test
- 1.9. PERFORMANCE OF THE TEST
- 1.9.1. Test conditions
- 1.9.2. Treatment and application of test substance
- 1.9.3. Test duration and sampling
- 1.9.4. Optional preliminary test
- 1.9.5. Measurements and analysis
- 2. DATA
- 2.1. TREATMENT OF RESULTS
- 3. REPORTING
- 3.1. TEST REPORT
- 4. REFERENCES
- Appendix 1
- GUIDANCE ON THE AEROBIC AND THE ANAEROBIC TEST SYSTEMS
- Aerobic test system
- Anaerobic test system
- Appendix 2
- EXAMPLE OF A GAS FLOW-THROUGH APPARATUS
- Appendix 3
- EXAMPLE OF A BIOMETER APPARATUS
- Appendix 4
- EXAMPLE CALCULATION FOR APPLICATION DOSE TO TEST VESSELS
Feedback