1,6-Bis(methoxybenzoyloxy)hexane

Endpoint summary

Description of key information

Based on the results of this study and in the absence of functional or morphological disturbances supporting the higher motor activity recordings for individual females at 150 mg/kg/day, a No Observed Adverse Effect Level (NOAEL) for Setafix X 11342 of 150 mg/kg/day was established.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20-02-2004 to 18-03-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study includes data generated according to generally valid and internationally accepted testing guidelines and performed according to GLP. The study was based on the following guideliens: -EC Directive 96/54/EEC, B.7 Repeated Dose (28 days) Toxicity (oral), 1996 -OECD 407, Repeated Dose 28-day Oral Toxicity Study in Rodents, 1995 -OPPTS 870.3050, Repeated dose 28-day Oral Toxicity Study in Rodents. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712-C-00-366,200.

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OPPTS 870.3050, Repeated dose 28-day oral toxicity study in rodents.

Deviations:

no

GLP compliance:

yes

Remarks:

22 July 2004 and 26 July 2004

Limit test:

no

Species:

rat

Strain:

other: Wistar Crl:(WI) BR (outbred, SPF-Quality)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 6 weeks
- Weight at study initiation: approx. 213 g (Male) and approx. 170 g (Female)
- Housing: Group housing of 5 animals per sex per cage in stainless steel suspended cages with wire mesh floors (55 x 34 x 21.5 cm height). During activity monitoring, animals were individually housed overnight in Macrolon plastic cages (type III, height 15 cm) with sterilised sawduct (woody-Clean type 3/4, Tecnilab-BMI BV, Someren, The Netherlands) provided as bedding.
- Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (from Altromin (code VRF-1, Lage, Germany). Each batch is analysed for nutrients and contaminants are analysed on regular basis. Results of analysis for ingredients and/or contaminants of diet, sawdust were assessed and did not reveal any findings that were considered to have affected study integrity.
- Water (e.g. ad libitum): Free access to tap water. Results of analysis for water were assessed and did not reveal any findings that were considered to have affected study integrity.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 (actual range 19.0 - 25.3)
- Relative Humidity (%): 30 - 70 (actual range: 32 - 58%)
- Air changes (per hr): approx. 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day

IN-LIFE DATES: From: 20 February 2004 To: 18 March 2004

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Method of formulation: Formulations (w/w) were prepared daily within 4 hours prior to dosing (within approximately 5 hours on day 1) and were homogenised to a visually acceptable level. Adjustment was made for specific gravity of vehicle. The solutions were stored at ambient temperature.
Stability of test substance in vehicle was determined to be at least 4 hours (determined at NOTOX)

VEHICLE
- Vehicle: propylene glycol, specific gravity 1.03628 days
- Justification for use and choice of vehicle (if other than water): Rationale for vehicle was ased on trial formulations performed at NOTOX
- Concentration in vehicle: Analysis of the accuracy of dose preparations revealed values within the range of 96 - 102 % of nominal, which was considered to represent an acceptable level of accurancy for formulations of this type.
- Amount of vehicle (if gavage): 5 ml/kg body weight. Actual dose volumes were calculated weekly according to the latest body weight.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of formulations prepared on day 14 were analysed to check homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 4 hours was also determined (highest and lowest concentration). The nalytical method used was based on the results of a separate project for the development and validation of the analytical method (NOTOX project 331008).
Test substance formulations in propylene glycol were noted as stable for at least 4 hours and formed a homogeneous suspension at the concentrations tested. Analysis of the accuracy of dose preparations revealed values within the range of 96 - 102 % of nominal, which was considered to reresent an acceptable level of accurancy for formulations of this type.

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily for at least 28 days , 7 days per week, approximately the same time as each day with a maximum of 4 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to necropsy.

Remarks:

Doses / Concentrations:
0 (vehicle)
Basis:
other: Vehicle

Remarks:

Doses / Concentrations:
50 mg/kg/day
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
150 mg/kg/day
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
1000 mg/kg/day
Basis:
nominal in diet

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
The dose levels were selected on the basis of a 5-day dose range finding study (NOTOX Project 399061). based on the results of this range finding study, dose levels selected for the main study (28 days toxicicty study) were: 50, 150 and 1000 mg/kg body weight/day

- Rationale for animal assignment:
20 males, 20 females (females were nulliparous and non-pregnant)
Animals were randmly assigned prior to comencement of treatment, by computer-generated random algorithm according to body weight, with all animals within ± 20 % of sex mean.

Positive control:

Not applicable

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once prior to start of treatment and on a weekly basis thereafter.

MORTALITY/VIABILITY
At least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily, detailed clinical observations were made in all animals. Once prior to start of treatment and on a weekly basis thereafter, this was also performed outside the home cage in a standard arena. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 1: grade 0 = absent, grade 1 = present
Maximum grade 3 or 4: grade 1 = slight, grade 2 = moderate, grade 3 = severe, grade 4 = very severe

NEUROBEHAVIOURAL EXAMINATION: Yes
- During week 4 of treatment, the following tests were performed on all animals:
- Hearing ability, Pupillary reflex, Static righting reflex and Grip strength (Score 0 = normal/present, score 1 = abnormal/absent)
- Motor Activity Test (recording period: 12 hours during overnight for individual animals). Motor activity measurements for males commenced on the last day of week 3.

BODY WEIGHT: Yes
- Time schedule for examinations: On days 1, 8, 15, 22 and 28

FOOD CONSUMPTION: Weekly

WATER CONSUMPTION: No
- Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled post mortem examination, between 7.30 and 10.30 a.m
- Anaesthetic used for blood collection: Yes, under iso-flurane anaesthesia
- Animals fasted: Yes (overnight, with a maximum of 20 hours)
- How many animals: 40
- Parameters examined: Erythrocytes count, Haemoglobin, Haematocrit, Mean corpuscular volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, Platelet count, Red cell distribution width, Differential leucocyte count (neutrophils, lymphocytes, monocytes, eosinophils, basophils).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled post mortem examination, between 7.30 and 10.30 a.m
- Anaesthetic used for blood collection: Yes, under iso-flurane anaesthesia
- Animals fasted: Yes (overnight, with a maximum of 20 hours)
- How many animals: 40
- Parameters examined: Alanine aminotransferase, alkaline phosphatase, Aspartate aminotransferase, total Bilirubin, Chloride, total Cholesterol, Creatine, Glucose, Phosphorus, total protein, Protein albumin, Urea, Calcium, Potassium and Sodium.

URINALYSIS: No data

CLOTTING POTENTIAL: Yes.
Parameters examined: Prothrombine, Partial throboplastin time

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

Necropsy:
All animals survivinf to the end of the observation period were deeply anaesthetised using iso-flurane and subsequently exsanguinated. All animals assigned to the study were necropsied and descriptions of all macroscopic abnormalities recorded.

Organs weights:
The following organ weights (and terminal body weight) were recorded from the animals on the scheduled day of necropsy: Adrenal glands, Brain, Liver, Heart, Kidneys, Spleen, Testes, Thymus and Epididymides

HISTOPATHOLOGY: Yes
The following slides were examined by a pathologist:
- all tissues and organs collected at the scheduled sacrifice from all animals of the control and the highest dose group
- all gross lesions of all animals
All abnormalities were described and documented.

Statistics:

The following statistical methods were used to analyse the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one test) based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex. The Student's t-test was applied for motor activity data.
- The Steel-test (many-to-one rank test) was applied when the data could not be assumed to follow a normal distribution.
- The exact Fisher-test was applied to frequency data.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before before printing.. Therefore, two groups may display the same printed means for a given parameter, yet dispaly different test statistics values.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Weight gains of high dose males were significantly lower than those of male controls on days 8 and 22 (p< 0.05). Body weights and body weight gain of other groups of treated animals remained in the same range as controls over the 4-week study period.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consymption before or after allowance for body weight was similar between treated and control animals.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Description (incidence and severity):

Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

slight chnages observed were considered to be of no toxicological significance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No dose-related trend was apparent and any deviations were considered to be of no toxicological relevance.

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No toxicologically significant changes were noted

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No toxicologically significant changes were noted

Gross pathological findings:

no effects observed

Description (incidence and severity):

No toxicologically relevant changes were noted

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No toxicologically significant changes were noted

Histopathological findings: neoplastic:

not specified

Details on results:

MORTALITY
No mortality occurred during the study.

CLINICAL SIGNS
There were no clinical signs over the 28-day observation period that were considered to be related to treatment.

Incidental findings that were noted included watery discharge from and/or dullness of the eys, alopecia on the neck, broken tail apex, diarrhoea and chromodacryorrhoea. These findings are occasionally noted in rats of this age and strain which are housed and treated under the conditions in this study. At incidence observed, these were considered signs of no toxicological significance. No clinical signs were shown by males at 150 mg/kg/day, females at 1000 mg/kg/day and control females.

NEUROBEHAVIOUR
Increased motor activity recordings were obtained for three high dose males (nos. 17, 18 and 20) and two high dose females treated with Setafix X 11342, when compared to control animals. Motor activity data between other treated and control animals were similar.

BODY WEIGHT AND WEIGHT GAIN
Slightly lower total body weight gains were noted in both genders at the high dose. Weight gains of high dose males were significantly lower than those of male controls on days 8 and 22 (p < 0.05).
Body weights and body weight gain of other groups of treated animals remined in the same range as conctrols over the 4-week study period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Food consumption before or after allowance for body weight was similar between treated and control animals.

WATER CONSUMPTION
Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

HAEMATOLOGY
A higher prothrombin time (PT) was measured in high dose males (p < 0.05). However, the control mean was slightly low and the mean was within the normal range of values for this type of study. The statistically lower partial thromboplastin time (PTT) in males at 50 mg/kg/day occurred in the absence of a treatment-related distribution. Therefore, this change was consiered to be of no toxicological significance. No further differences were noted in haematology parameters between treated and control rats.

CLINICAL CHEMISTRY
The following deviations in clinical biochemistry parameters distinguished high dose females from their controls:
- Increased alanine aminotransferase activity (ALAT; p < 0.05);
- Increased alkaline phosphatase activity (ALP; non-significant);
- Increased urea levels in female nos. 37 and 40 (p < 0.05);
- Increased glucose level in female no. 37 ( mean non-significant);
- Slightly increased inorganic phospate level (non-significant);
- Lower total protein level (non-significant).

The statistically significant lower total protein and albumin levels among males at 50, 150 and/or 1000 mg/kg/day were considered to be related to the slightly high control levels of these parameters. Also, no dose-related trend was apparent. In one high dose male (no. 19) a very low platelet count was measured. Since other animals of this dose group displayed normal values for this parameter, this was considered to have occurred incidentally. Therefore, these deviations were considered to be of no toxicological relevance.

ORGAN WEIGHTS
No toxicologically significant changes were noted regarding organ weights and organ:body weight ratios.
Slight changes in organ weights of high dose males (e.g. liver weights) were considered to be related to slightly lower terminal body weights. These changes were therefore considered not to be a sign of toxicity.

GROSS PATHOLOGY

- Macroscopic Examination:
Necropsy did not reveal any toxicologically relevant alterations.
Incidental findings among control and/or treated animals included enlargement of mandibular lymp nodes, pelvic dilation of the kidneys, a fractured tip of the tail, diaphragamatic hernia of the left lateral lobe of the liver, fluid in the uterus and reddish discolouration of the thymus. These findings are occasionally seen among rats used in these types of studies. In the absence of a treatment-related distribution they were considered changes of no toxicological significance.
No necropsy findings were noted in females at 50 mg/kg/day and above.

- Microscopic Examination:
There were no microscopic findings recorded which could be attributed to treatment with the test substance.
All microscopic findings were within the range of background pathology encountered in Wistar rats of this age and strain and occurred at similar incidencies and severity in both control and treated rats.

Dose descriptor:

NOAEL

Effect level:

150 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Critical effects observed:

not specified

RESULTS:

Accuracy, homogeneity and stability over 4 hours of formulation of test substance in propylene glycol were demonstrated by analyses.

Findings that distinguished treated from control animals included:

50 mg/kg/day:

No treatment-related findings noted

150 mg/kg/day:

Increased individual motor activity recordings

1000 mg/kg/day:

- Increased individual motor activity recordings

- Slightly low weight gain

- Increased prothrombin time

- Deviations in clinical biochemistry parameters in females included:

- Increased alanine aminotransferase activity;

- Increased alkaline phosphatase activity;

- Increased urea levels in individual females;

- Increased glucose level in one female;

- Slightly increased inorganic phosphate level;

- Lower total protein level.

Conclusions:

Based on the results of this study and in the absence of functional or morphological disturbances supporting the higher motor activity recordings for individual females at 150 mg/kg/day, a No Observed Adverse Effect Level (NOAEL) for Setafix X 11342 of 150 mg/kg/day was established.

Executive summary:

Subacute 28 -day oral toxicity with Setafix X 11342 by daily gavage in the rat.

The nature and purpose of this sub-acute toxicity study was to assess the toxic potential of the test substance when administered to rats by oral gavage for a period of 28 days. A No Observed Adverse Effect Level (NOAEL) was evaluated. The study should provide a part of a rational basis for toxicological risk assessment in man. The oral route was selected as it is a possible route of human exposure during manufacture, handling or use of the test substance.

Based on the results of a 5 -day range finding study, the dose levels for the 28 -day toxicity study were selected to be 0, 50, 150 and 1000 mg/kg/day.

The study was based on the following guidelines:

- EC Directive 96/54/EEC, B.7 Repeated Dose (28 days) Toxicity (oral), 1996.

- OECD 407, Repeated Dose 28 -day Oral Toxicity Study in Rodents, 1995

- OPPTS 870.3050, Repeated Dose 28 -day Oral Toxicity Study in Rodents. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712 -C-00 -366, 2000.

The test substance was administered daily for 28 days by oral gavage to SPF-bred Winstar rats.

One control group and three treated groups were tested, each consisting of 5 males and 5 females.

The following parameters were evaluated: clinical signs daily; functional observation tests; body weight and food consumption weekly; clinical pathology and mcroscopy at termination; organ weights and histopathology on a selection of tissues.

No in-life observations indicative of toxicity were obtained except for a slightly low weight gain for high dose animals during treatment. This change was, however, not accompanied by concurrent changes in food intake. Also, increased motor activity recordings were obtained for individual high dose males, and females at 150 and 1000 mg/kg/day. However, concurrent and similar changes of the low or high sensors were not always apparent and there was no supportive clinical signs of hyperactivity. A relationship to treatment was considered to be uncertain.

There were no changes at determination of clinical appearance, performance of functional observations, food consumption measurements, macroscopic examination, organ weight determination and microscopic examination that were considered to be an effect of treatment.

Based on the results of this study and in the absence of functional or morphological disturbances supporting the higher motor activity recordings for individual females at 150 mg/kg/day, a No Observed Adverse Effect Level (NOAEL) for Setafix X 11342 of 150 mg/kg/day was established.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

150 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The study includes data generated according to generally valid and internationally accepted testing guidelines and performed according to GLP.
The study was based on the following guidelines:
- EC Directive 96/54/EEC, B.7 Repeated Dose (28 days) Toxicity (oral), 1996.
- OECD 407, Repeated Dose 28 -day Oral Toxicity Study in Rodents, 1995
- OPPTS 870.3050, Repeated Dose 28 -day Oral Toxicity Study in Rodents. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712 -C-00 -366, 2000.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Study not available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Study not availble

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

exposure considerations

Justification for data waiving:

a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Study not available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Study not available

Additional information

Subacute 28 -day oral toxicity with Setafix X 11342 by daily gavage in the rat.

The nature and purpose of this sub-acute toxicity study was to assess the toxic potential of the test substance when administered to rats by oral gavage for a period of 28 days. A No Observed Adverse Effect Level (NOAEL) was evaluated. The study should provide a part of a rational basis for toxicological risk assessment in man. The oral route was selected as it is a possible route of human exposure during manufacture, handling or use of the test substance.

Based on the results of a 5 -day range finding study, the dose levels for the 28 -day toxicity study were selected to be 0, 50, 150 and 1000 mg/kg/day.

The study was based on the following guidelines:

- EC Directive 96/54/EEC, B.7 Repeated Dose (28 days) Toxicity (oral), 1996.

- OECD 407, Repeated Dose 28 -day Oral Toxicity Study in Rodents, 1995

- OPPTS 870.3050, Repeated Dose 28 -day Oral Toxicity Study in Rodents. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712 -C-00 -366, 2000.

The test substance was administered daily for 28 days by oral gavage to SPF-bred Winstar rats.

One control group and three treated groups were tested, each consisting of 5 males and 5 females.

The following parameters were evaluated: clinical signs daily; functional observation tests; body weight and food consumption weekly; clinical pathology and microscopy at termination; organ weights and histopathology on a selection of tissues.

RESULTS:

Accuracy, homogeneity and stability over 4 hours of formulation of test substance in propylene glycol were demonstrated by analyses.

Findings that distinguished treated from control animals included:

50 mg/kg/day:

No treatment-related findings noted

150 mg/kg/day:

Increased individual motor activity recordings

1000 mg/kg/day:

- Increased individual motor activity recordings

- Slightly low weight gain

- Increased prothrombin time

- Deviations in clinical biochemistry parameters in females included:

- Increased alanine aminotransferase activity;

- Increased alkaline phosphatase activity;

- Increased urea levels in individual females;

- Increased glucose level in one female;

- Slightly increased inorganic phosphate level;

- Lower total protein level.

CONCLUSION

No in-life observations indicative of toxicity were obtained except for a slightly low weight gain for high dose animals during treatment. This change was, however, not accompanied by concurrent changes in food intake. Also, increased motor activity recordings were obtained for individual high dose males, and females at 150 and 1000 mg/kg/day. However, concurrent and similar changes of the low or high sensors were not always apparent and there was no supportive clinical signs of hyperactivity. A relationship to treatment was considered to be uncertain.

Blood analyses revealed slight deviations in clinical biochemistry parameters in high dose females (alanine aminotransferase activity and urea levels were significantly higher). There were no supportive microscopic lesions, and a relationship of the deviations to treatment with the best test substance was considered to be uncertain.

The higher prothrombin time in high dose males was within the normal range of values for this type of study and the control mean was slightly low. Also, there was no histopathologic evidence of liver dysfunction. Therefore, this change was considered to be of no toxicological significance.

There were no changes at determination of clinical appearance, performance of functional observations, food consumption measurements, macroscopic examination, organ weight determination and microscopic examination that were considered to be an effect of treatment.

Based on the results of this study and in the absence of functional or morphological disturbances supporting the higher motor activity recordings for individual females at 150 mg/kg/day, a No Observed Adverse Effect Level (NOAEL) for Setafix X 11342 of 150 mg/kg/day was established.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The nature and purpose of this sub-acute toxicity study was to assess the toxic potential of the test substance when administered to rats by oral gavage for a period of 28 days. A No Observed Adverse Effect Level (NOAEL) was evaluated. The study should provide a part of a rational basis for toxicological risk assessment in man. The oral route was selected as it is a possible route of human exposure during manufacture, handling or use of the test substance.
The study was based on the following guidelines:
- EC Directive 96/54/EEC, B.7 Repeated Dose (28 days) Toxicity (oral), 1996.
- OECD 407, Repeated Dose 28 -day Oral Toxicity Study in Rodents, 1995
- OPPTS 870.3050, Repeated Dose 28 -day Oral Toxicity Study in Rodents. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712 -C-00 -366, 2000.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
According to COLUMN 2 Specific Rules For Adaptation From COLUMN 1 set out in Section 8.6.1 Annex VIII of REACH the short-term toxicity study (28 days): INHALATION does not need to be conducted if relevant human exposure can be excluded in accordance with Annex XI section 3.

For the test substance SETAFIX X 11342 exposure of humans via inhalation is not the most relevant route of human exposure especially considering that the test substance SETAFIX X 11342 is a solid and its approximate vapour pressure 0.019 Pa at 20 °C is relatively low and the possibility of exposure to aerosols, particles or droplets of an inhalable size is likely to vary from minimal to negligible.

Therefore, testing in accordance with sections 8.6 and 8.7 of Annex VIII and Annex IX may be omitted

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
According to COLUMN 2 Specific Rules For Adaptation From COLUMN 1 set out in Section 8.6.1 Annex VIII of REACH the short-term toxicity study (28 days): INHALATION does not need to be conducted if relevant human exposure can be excluded in accordance with Annex XI section 3.

For the test substance SETAFIX X 11342 exposure of humans via inhalation is not the most relevant route of human exposure especially considering that the test substance SETAFIX X 11342 is a solid and its approximate vapour pressure 0.019 Pa at 20 °C is relatively low and the possibility of exposure to aerosols, particles or droplets of an inhalable size is likely to vary from minimal to negligible.

Therefore, testing in accordance with sections 8.6 and 8.7 of Annex VIII and Annex IX may be omitted

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
According to COLUMN 2 Specific Rules For Adaptation From COLUMN 1 set out in Section 8.6.1 Annex VIII of REACH the short-term toxicity study (28 days):DERMAL does not need to be conducted if relevant human exposure can be excluded in accordance with Annex XI section 3.

Based on all the available information, for the test substance SETAFIX X 11342, exposure of humans via dermal route is not the most relevant route of human exposure especially considering that the physico-chemical properties of the test substance SETAFIX X 11342, a solid, do not suggest potential for a significant rate of absorption through the skin and hence the possibility of dermal absorption is likely to be negligible.

Therefore, testing in accordance with sections 8.6 and 8.7 of Annex VIII and Annex IX may be omitted

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
According to COLUMN 2 Specific Rules For Adaptation From COLUMN 1 set out in Section 8.6.1 Annex VIII of REACH the short-term toxicity study (28 days):DERMAL does not need to be conducted if relevant human exposure can be excluded in accordance with Annex XI section 3.

Based on all the available information, for the test substance SETAFIX X 11342, exposure of humans via dermal route is not the most relevant route of human exposure especially considering that the physico-chemical properties of the test substance SETAFIX X 11342, a solid, do not suggest potential for a significant rate of absorption through the skin and hence the possibility of dermal absorption is likely to be negligible.

Therefore, testing in accordance with sections 8.6 and 8.7 of Annex VIII and Annex IX may be omitted

Justification for classification or non-classification

No in-life observations indicative of toxicity were obtained except for a slightly low weight gain for high dose animals during treatment. This change was, however, not accompanied by concurrent changes in food intake. Also, increased motor activity recordings were obtained for individual high dose males, and females at 150 and 1000 mg/kg/day. However, concurrent and similar changes of the low or high sensors were not always apparent and there was no supportive clinical signs of hyperactivity. A relationship to treatment was considered to be uncertain.

There were no changes at determination of clinical appearance, performance of functional observations, food consumption measurements, macroscopic examination, organ weight determination and microscopic examination that were considered to be an effect of treatment.

Based on the results of this study and in the absence of functional or morphological disturbances supporting the higher motor activity recordings for individual females at 150 mg/kg/day, a No Observed Adverse Effect Level (NOAEL) for Setafix X 11342 of 150 mg/kg/day was established.

Therefore no 'significant' toxic effects were seen and therefore classification under 1272/2008 is not warranted based on this study.