4,4'-Methylene-bis-(3-chloro-2,6-diethylaniline)

Endpoint summary

Description of key information

Subacute 28-day oral toxicity (feeding):  

 

The study was performed 1987 as GLP-test following EC-test method B.7 on Sprague-Dawley rats. Application of test material was via diet, the dose levels were 0, 100, 300 and 1000 ppm. No deaths and no clinical signs of toxicity were noted. Laboratory findings included increased platelet counts in both sexes in the top dose group and slightly increased serum calcium in top dose females. Absolute and relative liver and adrenal weights were increased in both sexes in the top dose group only. The only effect seen microscopically was hypertrophy of the centrilobular cells of the liver. In conclusion, the NOEL was found to be 300 ppm (= 38 mg/kg/day).  

  

Subchronic 90-day oral toxicity (feeding): 

 

The study was performed 1996 as GLP-test following OECD-test method 408 on Sprague-Dawley rats. Application of test material was via diet, the dose levels were 0, 100, 300 and 1600 ppm. There were two unscheduled deaths, but no clinical signs of toxicity observed. There were effects observed on body weights, food efficiency haematology, biochemistry, organ weights and in macroscopic and microscopic pathology. The findings were noted mainly in the mid and high dose groups. No effects were seen on food consumption, water consumption, ophthalmoscopy and urinalysis. The microscopical findings were still present after a 4-week recovery period in females previously given 1600 ppm but there the incidences were not statistically significant from those of control rats.  

  

Repeated dose inhalation toxicity: 

 

The use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Furthermore, the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated inhalation route administration. 

  

Repeated dose dermal toxicity: 

 

The substance is unlikely to be inhaled, skin contact is unlikely and the physicochemical and toxicological properties suggest low potential for significant rate of absorption through the skin. Furthermore, the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated dermal route administration.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September - December 1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: stable

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, F.R.G., Germany
- Weight at study initiation: body weights of the males ranged from 142 to 162 g and those of the females from 111 to 137 g
- Housing: gang housing (6 animals/sex/group/cage) in polycarbonate cages
- Diet: Standard laboratory diet (RMH-0. pellet diameter 9 mm, Hope Farms, Woerden,
- Water: ad libitum
- Acclimatisation: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 21°C
- Humidity: mean 60-70%
- Lightning: artificial light sequence was 12 hours light (7:00 to 19:00), 12 hours dark (19:00 to 7:00).

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Remarks:

mixed with diet

Details on oral exposure:

Method of administration: feeding test (test item mixed with diet); for diet preparation, see "any other information on materials and methods"

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Based on analysis of the stability of the test substance in the diets of the pilot study, it was decided to prepare the diets for the 28-day study only once, i.e. 1 day prior to study start. The sequence for diet mixing and pelleting was control group (no test substance), followed by low dose, medium dose and high dose group. The weighed out test substance was mixed with a fraction of the diet in a Kenwood mixer for 6 minutes resulting in a 3% premix. In a professional dough mixer the appropriate amount of diet was moistened with 7% (w/w) fresh tap-water and was subsequently supplemented by the premix.

DIET PREPARATIONS:
Dosage (ppm) Diet (kg) Test substance (g)
0 25 0
100 25 2.5
300 25 7.5
1000 25 25.0

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

0 ppm

Remarks:

= 0 mg/kg bw/day nominal in diet

Dose / conc.:

100 ppm

Remarks:

= 100 mg/kg bw/day nominal in diet

Dose / conc.:

300 ppm

Remarks:

= 300 mg/kg bw/day nominal in diet

Dose / conc.:

1 000 ppm

Remarks:

= 1000 mg/kg bw/day nominal in diet

No. of animals per sex per dose:

MALES:
- 6 animals at 0 ppm
- 6 animals at 100 ppm
- 6 animals at 300 ppm
- 6 animals at 1000 ppm

FEMALES:
- 6 animals at 0 ppm
- 6 animals at 100 ppm
- 6 animals at 300 ppm
- 6 animals at 1000 ppm

Control animals:

yes, plain diet

Positive control:

no positive control tested

Observations and examinations performed and frequency:

Cage-side observations:
With the exception of days 0, 7, 21 and 28, cage-side observations were performed once daily on working days (beginning on day 1) as well as weekends until terminal sacrifice on day 28. Any deviations from normal were recorded. In particular, attention was paid to changes of skin, fur, eyes, mucous membranes, respiratory system. faeces and general appearance. On working days a mortality check was performed in the eveni

Physical examinations:
Once a week, i.e. on days 0 (prior to dosing), 7, 14, 21 and 28 the animals underwent a physical examination. In addition to the parameters mentioned for cage-side observations, particular attention was paid to ears. mouth, urogenital region, anus, abnormal masses, gait, general state and behaviour.

Body weights and food consumption
Individual body weights were determined immediately prior to the first dosing (day 0), weekly thereafter (days 7, 14, 21 and 27), and on day 28 prior to sacrifice (fasted body weight). Individual food consumption was recorded weekly (days 7, 14, 21 and 27) and was expressed as grams food consumed per animal and as grams food consumed per 100 grams body weight per day.

Haematology and clinical chemistry
Prior to sacrifice on day 28 the animals were anaesthetized with ether and the abdominal cavity was opened. Blood was collected via the abdominal aorta by means of an infusion set provided with a winged needle (Mediwing, Argyle. Sherwood Medical Industries, Tullamore, Ireland). For haematology approximately 1.5 ml of blood were collected directly into vials containing 0.03 ml of di- and tripotassium EDTA (30% (W/V), pH 7.4). Immediately thereafter blood samples were collected in clot tubes for clinical chemistry. The vials intended for haematology were rotated at 90 rpm until transportation to 'Bergschot Centrum voor Onderzoek CBCO)' in Breda. The Netherlands for analysis. The maximum period between blood collection and transport to BCO was 5.5 hours. The approximately 3 ml blood samples intended for clinical chemistry were allowed to clot during 30-60 minutes. The serum was separated by centrifugation and transferred into another tube. Serum samples were kept on ice until analysis by BCO.
The red blood cells of the samples were counted using a "Coulter" blood cell counter, while their size distribution was determined using the "Haemalog 6000" (Technicon). Measurement of haemoglobin concentration, counting of white blood cells and subsets thereof, i.e. lymphocytes, neutrophils, large unstained cells, monocytes, eosinophils and basophils, and the counting of thrombocytes were also performed using the Haemalog 6000. The haematocrit values were determined using a microcentrifuge. All clinical chemical parameters were measured in serum samples using a "Parallel" clinical chemistry analyzer.

Sacrifice and pathology:

On completion of blood collection the animals were sacrificed by exsanguination. Subsequently a full gross necropsy was performed which included examination of the external surface and openings, and the cranial, thoracic and abdominal cavities with their contents. Liver, adrenals, kidneys, spleen and testes were removed and weighed.
The following tissues were removed and preserved in 10% formalin: Liver. spleen, kidneys, adrenals. heart and macroscopically abnormal tissues. Fixed organs and tissues from animals of the control and high dose group were embedded in paraffin, sectioned, stained with haematoxylin and azophloxine, and examined microscopically.

Statistics:

Means with standard deviation were calculated for all quantitative parameters. Statistical procedures were carried out for quantitative data suspected of changes and were based on a one-way analysis of variance supplemented with a t-test (Biodata Handling with Microcomputers, Barlow. 1983). One-way analysis of variance (F-test) was used to analyse the results for overall effects of dosage. Significant differences between control and individual dosages were also assessed by using the F-test where P < 0.05 was accepted as the lowest level of significance . The total food consumption represents the group mean of the sum of the individual food consumption over the entire period (day 7, 14. 21 and 27). whereas the body weight gain was obtained by calculating the group mean of the mean individual body weight gain (obtained from a linear regression analysis for the individual body weight over the entire period, i.e. day 0, 7, 14, 21 and 27).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

With respect to the group mean haematology data no evident test substance related effect was observed, except for a statistically significant (P<0.05) increase of thrombocytes in the high dose group (i.e. 18% and 20% increase for males and females, respectively). The remaining haemometric values were comparable for the control group and the treatment groups. In addition, the obtained values were in general agreement with those reported in the literature (Charles River Laboratories. Technical Bull~tin, 1984). Incidental abnormal values were obtained with the following animals. Animals A4 and C4 showed an increased haematocrit value: the latter animal also had an increased number of erythrocytes. Animal B8 showed an increase of erythrocytes and haemoglobin and decreased mean cellular volume (MCV) and MCV/RBC values. With respect to the group mean clinical chemistry data no
statistically significant test substance related effect was observed in sera of male or female animals, with the exception of the calcium concentration that was increased in female animals of the high dose group (6% increase; P=0.05). The obtained values were in general agreement with those reported in the literature. Serum aspartate aminotransferase and alanine aminotransferase were increased in animals A11 and C10.

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Absolute and relative liver and adrenal weights were increased in both sexes in the top dose group only.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Macroscopic examination of all animals at necropsy revealed no evident test substance related gross abnormalities. with exception of dilated renal pelvis (observed once in the low dose group and twice in the high dose group; considered as a congenital abnormality in this rat strain), petechiae of the stomach (observed once in the control group), enlarged adrenals (observed once in low dose group and twice in the high dose group) and petechiae of the thymus (observed once in the high dose group). With the exception of enlarged adrenals in animals of the high dose group, the above findings were considered incidental and not test substance related.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Microscopic examination of the liver of animals dosed at 1000 ppm revealed hypertrophy of centrilobular liver cells.

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

Details on results:

SUMMARY:

CLINICAL OBSERVATIONS:
No deaths and no clinical signs of toxicity.

LABORATORY FINDINGS:
Platelet counts were increased in both sexes in the top dose group and serum calcium was slightly increased in top dose females.

EFFECTS IN ORGANS:
Absolute and relative liver and adrenal weights were increased in both sexes in the top dose group only. The only effect seen microscopically was hypertrophy of the centrilobular cells of the liver.

Key result

Dose descriptor:

NOEL

Effect level:

300 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

haematology

Key result

Critical effects observed:

no

Lowest effective dose / conc.:

100 ppm

Organ:

bladder

Conclusions:

Based on the findings it was concluded that a dietary exposure level of 300 ppm (equivalent to 38 mglkglday for males and females combined) represents the No Effect Level for P5367 in the present study.

Executive summary:

The study was performed 1987 as GLP-test following EC-test method B.7 on Sprague-Dawley rats. Application of test material was via diet, the dose levels were 0, 100, 300 and 1000 ppm. No deaths and no clinical signs of toxicity were noted. Laboratory findings included increased platelet counts in both sexes in the top dose group and slightly increased serum calcium in top dose females. Absolute and relative liver and adrenal weights were increased in both sexes in the top dose group only. The only effect seen microscopically was hypertrophy of the centrilobular cells of the liver. In conclusion, the NOEL was found to be 300 ppm (= 38 mg/kg/day).

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Margate, Kent, UK
- Weight at study initiation: in the range 181 g to 234 g for males and 146 g to 185 g for females
- Housing: gang housing (5 animals/sex/group/cage) in polycarbonate cages
- Diet: pelleted SDS Rat and Mouse No. 1 modified maintenance diet
- Water: ad libitum
- Acclimatisation: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 -24°C
- Humidity: 32 - 71%
- Lightning: artificial light sequence was 12 hours light, 12 hours dark

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
Treated diets were prepared once every two weeks. A pre-mix was prepared by grinding the test substance directly into untreated basal diet and mixing in a Turbula mixer for a minimum period of 5 minutes. The required concentrations were then prepared by direct dilution of the pre-mix with further quantities of untreated diet, homogeneity being achieved in a turbula mixer for a period of 5 minutes.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

FORMULATION SAMPLING AND ANALYSIS:
Prior to the commencement of the study the proposed formulation procedure was checked by chemical analysis to confirm that the method was acceptable and that the homogeneity and stability of the formulation was satisfactory under the conditions of the study. Samples of the formulation(s) for Weeks 1 and 11 were also analysed to check the accuracy of preparation. In Week 13 samples were stored at room temperature for possible future analysis and were discarded after the Study Director was satisfied that the results from the Weeks 1 and 11 analyses were satisfactory.

HPLC-ANALYSIS:

1) Sample extraction:
A representative sub-sample (10 g) of test diet formulation, obtained by a standard riffling technique, was extracted (mechanical shaking, 30 minutes) using methanol (160 ml). The extract was filtered (Whatman GP/A) and the filtrate was diluted using methanol. A suitable volume (10 ml) of the diluted extract was evaporated (RFE, 40°C) to dryness and the residue re-dissolved in mobile phase (10 ml) to provide a solution containing P5367 in the expected concentration range 2 - 4 ug/ml. The final solution was filtered (Whatman® PURADISCTM 25PP, 0.2 um) and the concentration of P5367 was quantified by high performance liquid chromatography using ultraviolet detection as detailed in the following section.

2) Typical chromatographic conditions
- Pump: Perkin Elmer LC250
- Autosampler: Perkin Elmer ISS200
- Detector: Applied Biosystems 785A
- Data handling: Perkin-Elmer 1020LC Plus
- Analytical column: LiChrosorb RP-18, 7 um, 125 x 4 mm id, Merck Ltd.
- Guard column: Aquapore ODS, 7 um, 15 x 3.2 mm id, Applied Biosystems Inc.
- Column temperature: Ambient, nominally 21°C ± 1°C
- Mobile phase: Methanol/aqueous O.lM ammonium formate (85/15 v/v)
- Flow rate: 1.0 ml/minute
- Detector wavelength: UV, 246 nm
- Injection volume: 20 ul
- Sensitivity: 35 mY
- Retention volume: 4 ml

Duration of treatment / exposure:

Test duration: 90 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

0 ppm

Remarks:

= 0 mg/kg bw/day nominal in diet

Dose / conc.:

100 ppm

Remarks:

= 100 mg/kg bw/day nominal in diet

Dose / conc.:

300 ppm

Remarks:

= 300 mg/kg bw/day nominal in diet

Dose / conc.:

1 600 ppm

Remarks:

= 1600 mg/kg bw/day nominal in diet

No. of animals per sex per dose:

MALES:
- 10 animals at 0 ppm
- 5 animals at 100 ppm
- 5 animals at 300 ppm
- 10 animals at 1600 ppm

FEMALES:
- 10 animals at 0 ppm
- 5 animals at 100 ppm
- 5 animals at 300 ppm
- 10 animals at 1600 ppm

Control animals:

yes, plain diet

Details on study design:

Following a total acclimatisation period of 18 days treatment continued until completion of 13 weeks. Following this selected animals from the control and high dose levels were maintained for 4 weeks untreated to assess recovery.

Positive control:

no positive control tested

Observations and examinations performed and frequency:

See "any other information on materials and methods"

Sacrifice and pathology:

On completion of 13 weeks of treatment, all surviving Main group rats were killed. Recovery group rats were killed after a 4 week period without treatment. As the terminal procedures took 2 days to complete, the Main group animals continued to receive treated diet until the day on which they were killed. The duration of the dosing period, however, is reported as 13 weeks. All rats were killed by carbon dioxide asphyxiation and subjected to the necropsy procedure indicated in the report.

Statistics:

All statistical analyses were carried out separately for males and females. Data relating to food and water consumption were analysed on a cage basis. For all other parameters, the analyses were carried out using the individual animal as the basic experimental unit. Food consumption data were analysed using cumulative cage totals and water consumption data were analysed as the total recorded intake over selected time periods, expressed on a daily/weekly basis. Bodyweight data were analysed using weight gains.

The following sequence of statistical tests was used for food consumption, water consumption, bodyweight, clinical pathology and organ weight data:
- If the data consisted predominantly of one particular value (relative frequency of the mode exceeded 75 %), the proportion of animals with values different from the mode was analysed, (Fisher, 1950 and Mantel, 1963). Otherwise:
- A test was applied to test for heterogeneity of variance between treatments, (Bartlett, 1937). Where significant (at the 1 % level) heterogeneity was found, a logarithmic transformation was tried to see if a more stable variance structure could be obtained.
- If no significant heterogeneity was detected (or if a satisfactory transformation was found), a one-way analysis of variance was carried out. If significant heterogeneity of variance was present, and could not be removed by a transformation, an analysis of ranks was used, (Kruskal-Wallis, 1952/3).
- Except for pre-dose data, analyses of variance were followed by Students 't' test and WiIliams test (WiIliams 197112) for a dose related response, although only the one thought most appropriate for the response pattern observed was reported. The Kruskal-Wallis analyses were followed by the non-parametric equivalents of these tests (Shirley, 1977)

Clinical signs:

no effects observed

Mortality:

mortality observed, treatment-related

Description (incidence):

There were two unscheduled deaths on the study, both control rats. Both rats were found dead immediately following blood sampling at the scheduled laboratory investigations and these deaths were considered likely to be a result of trauma related to the blood sampling procedures.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

A significant reduction in bodyweight gain was noted during Week 1 for females and, to a lesser extent, for males treated with 1600 ppm in comparison with concurrent controls. Thereafter, bodyweight gain for these groups was similar to that of the controls such that overall gain was similar to concurrent control for the males and still slightly reduced for the females in comparison with the control. The overall mean bodyweight gain for previously treated groups was marginally higher than concurrent controls during the recovery period.

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

The efficiency of food utilisation for rats of both sexes treated with 1600 ppm was marginally inferior to control during Week 1. Overall, mean food conversion ratios for all treated groups were similar to those of the controls.
The mean food conversion ratios for the recovery period for previously treated groups were essentially similar to those of the controls. Achieved intakes of test item as expected from fixed dietary inclusion levels, the achieved dietary intakes in terms of 'mg/kg/day' generally decreased as the bodyweight of the animals increased. Intervals between group mean achieved intakes generally reflected the intervals between dietary inclusion levels. Mean achieved intakes over the study as a whole were 7.6, 23.4 and 129 mg/kg/day in males and 8.5, 28.2 and 143 mg/kg/day in females

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

no effects observed

Description (incidence and severity):

A slight, not statistically significant, increase in overall mean water intake was noted for females treated with 1600 ppm in comparison with the concurrent controls during the Week 12 measurements.

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

There were slight deviations in several haematological parameters at 1600 ppm which were of uncertain relationship to treatment.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Significant increases in mean cholesterol and triglyceride levels were evident during Weeks 4 and 13 for females receiving 1600 ppm in comparison with controls. Increases in mean cholesterol and triglyceride levels were again noted at the end of the recovery phase investigations for previously treated females in comparison with the control, although these increases generally were of a lesser degree than at Week 13.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Statistically significant increases in group mean adjusted liver and spleen weights were noted at the terminal necropsy for male and female rats receiving 1600 ppm in comparison with the concurrent controls. A slight, and statistically significant, increase in group mean adjusted kidney weights was noted for females at 300 ppm and males and females at 1600 ppm in comparison with the controls (not dose related in females). A statistically significantly lower group mean adjusted liver weight was noted at the recovery kill for males previously treated with 1600 ppm. Statistically significantly higher group mean adjusted liver weight was noted for females previously receiving 1600 ppm in comparison with concurrent controls.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Pale punctate foci in the lungs was noted at the terminal and recovery kills in a small proportion of rats given 1600 ppm and in one female given 300 ppm compared with none in the control rats.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The frequency and degree of centrilobular hepatocyte hypertrophy in the liver was statistically significantly increased in rats of both sexes given 1600 ppm and in males given 300 ppm when compared to controls. Foamy alveolar macrophage aggregation(s) with or without cholesterol clefts were statistically significantly increased in rats of both sexes given 1600 ppm when compared to controls. No changes were seen to account for observed changes in spleen and kidney weight. After a 4 week recovery period the above findings were still present in females previously given 1600 ppm but there the incidences were not statistically significant from those of control rats.

Other effects:

not examined

Key result

Dose descriptor:

NOAEL

Effect level:

300 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

food efficiency

gross pathology

haematology

histopathology: neoplastic

organ weights and organ / body weight ratios

Key result

Dose descriptor:

NOEL

Effect level:

100 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

food efficiency

gross pathology

haematology

histopathology: neoplastic

organ weights and organ / body weight ratios

Key result

Critical effects observed:

no

Lowest effective dose / conc.:

100 mg/kg bw/day (nominal)

Conclusions:

The dosage of 1600 ppm P5367 (equivalent to 129 mg/kg/day in males and 143 mg/kg/day in females) was a clear toxic effect level. 100 ppm (equivalent to 7.6 mg/kg/day in males and 8.5 mg/kg/day in females) produced no evidence of toxic effect and represents a no effect level. Some of the effects seen at 1600 ppm were also present at 300 ppm.

Executive summary:

The study was performed 1996 as GLP-test following OECD-test method 408 on Sprague-Dawley rats. Application of test material was via diet, the dose levels were 0, 100, 300 and 1600 ppm. There were two unscheduled deaths, but no clinical signs of toxicity observed. There were effects observed on body weights, food efficiency haematology, biochemistry, organ weights and in macroscopic and micrscopic pathology. The findings were noted mainly in the mid and high dose groups. No effects were seen on food consumption,, water consumption, ophtalmoscopy and urinalysis. The microscopical findings were still present after After a 4 week recovery period in females previously given 1600 ppm but there the incidences were not statistically significant from those of control rats.

Laboratory findings included increased platelet counts in both sexes in the top dose group and slightly increased serum calcium in top dose females. Absolute and relative liver and adrenal weights were increased in both sexes in the top dose group only. The only effect seen microscopically was hypertrophy of the centrilobular cells of the liver.

In conclusion, the NOEL was found to be 300 ppm (= 38 mg/kg/day).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

38 mg/kg bw/day

Study duration:

subacute

Experimental exposure time per week (hours/week):

24

Species:

rat

Quality of whole database:

Klimisch 1

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

a sub-chronic toxicity study (90 days) does not need to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28-day 'limit test' and human exposure is limited

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur. Furthermore the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated inhalation route administration.

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

a sub-chronic toxicity study (90 days) does not need to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28-day 'limit test' and human exposure is limited

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur. Furthermore the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated inhalation route administration.

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

a short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration

a sub-chronic toxicity study (90 days) does not need to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28-day 'limit test' and human exposure is limited

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The substance is unlikely to be inhaled, skin contact is unlikely and the physicochemical and toxicological properties suggest low potential for significant rate of absorption through the skin. Furthermore the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated dermal route administration.

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

a short-term toxicity study does not need to be conducted because a reliable sub-chronic (90 days) or chronic toxicity study is available, conducted with an appropriate species, dosage, solvent and route of administration

a sub-chronic toxicity study (90 days) does not need to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28-day 'limit test' and human exposure is limited

other:

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
The substance is unlikely to be inhaled, skin contact is unlikely and the physicochemical and toxicological properties suggest low potential for significant rate of absorption through the skin. Furthermore the results of laboratory animal studies show low acute dermal toxicity. In the 28-day and 90-day repeated dose study via oral administration does not exacerbate systemic toxicity effects which suggest bioavailability is low, thereby there is low toxicity potential. This intrinsic property/toxicity potential can be extrapolated to repeated dermal route administration.

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the data available the substance is not classified or labeled according to Regulation 1272/2008/EC (CLP).