Fatty acids, tall-oil, reaction products with formaldehyde and (Z)-N-9-octadecenyl-1,3-propanediamine

Repeated dose toxicity: oral

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Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650

Deviations:

no

Principles of method if other than guideline:

In addition, the procedures described in the protocol were conform to:
- OECD Guideline 421
- EPA OPPTS 870.3550
- EU method B.7
- OECD Guideline 407
- EPA OPPTS 870.3050

Limit test:

no

Test material

Test material information

Constituent 1

Test animals

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY
At 1000 mg/kg, hunched posture, piloerection, rales and/or lean appearance were recorded for the females euthanized in extremis (nos. 75 and 79) in the days preceding their death. Piloerection was also noted incidentally for four of the surviving females at 1000 mg/kg between Day 20 post-coitum and lactation Day 4, and hunched posture was transiently observed for a single surviving female at this dose level. Salivation was noted for all animals at 1000 mg/kg (from Day 9 of the premating period onwards) and for a few males at 300 mg/kg. This was considered to be a physiological response rather than a sign
of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing). This sign may be related to the taste or possible irritancy of the test substance. For one male at 300 mg/kg a swelling (nodule) was noted on the chest from Day 2 of treatment onwards which increased in diameter from 5 mm to 31 mm (correlating to a carcinoma; see also section 7.2.8). It was not considered test substance related. Based on the observations performed, there was no sign for that this nodule impaired the normal functioning of this male. Other incidental findings noted included alopecia and scales. These findings occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At the incidence observed, these were considered signs of no toxicological significance.

BODY WEIGHT AND WEIGHT GAIN
Reduced body weights and body weight gains were recorded for males at 300 and 1000 mg/kg during the entire treatment period (not always statistically significant). Mean values for body weight and body weight gain were up to 8% and 67%, respectively, lower as compared to the concurrent control. For females at 1000 mg/kg, a trend towards slightly lower body weights and body weight gains from Day 14 of the post coitum period was noted. Since differences from controls were only slight and not statistically significant, this finding was not considered as toxicologically relevant.

FOOD CONSUMPTION
There were no toxicologically relevant effects on food consumption. An apparent dose related trend towards lower food intake was recorded for males during the first days of treatment. As the differences from controls were only slight and changes were transient, they were not considered to be toxicologically relevant.

HAEMATOLOGY
The following treatment related changes in haematology parameters were noted at 300 and/or 1000 mg/kg:
- Higher mean white blood cell counts for males and females at 1000 mg/kg (161% and 191% of controls, respectively). This was due to a higher mean absolute neutrophil count (392% and 391% of controls, respectively; not statistically significant for males).
- Higher mean absolute neutrophil counts for males and females at 300 mg/kg (283% and 209% of controls, respectively) without any concurrent increase in mean white blood cell count.
- Higher mean relative neutrophil counts for males and females at 300 mg/kg (201% and 157% of controls, respectively; not statistically significant for females) and 1000 mg/kg (216% and 189% of controls, respectively; not statistically significant for males).
- Increased mean prothrombin time (PT) for males at 1000 mg/kg (109% of control). Other (statistically significant) changes in haematological parameters were considered not to be of toxicological relevance, since values remained within the normal range of biological variation, changes occurred in the absence of a treatment related distribution and/or were due to a higher white blood cell count. These changes included lower mean relative lymphocyte counts at 300 mg/kg (males) and 1000 mg/kg (females), higher mean absolute monocyte counts at 300 mg/kg (males) and 1000 mg/kg (females), a lower mean relative eosinophil count at 1000 mg/kg (males), and a higher mean platelet count at 1000 mg/kg (females). Haematological parameters of treated rats were considered not to have been affected by treatment at 100 mg/kg.

CLINICAL CHEMISTRY
At 1000 mg/kg higher alanine aminotransferase activity (ALAT) was noted for males (184% of control). Any other statistically significant changes were considered to be of no toxicological significance as they occurred in the absence of a treatment-related distribution and remained within the range considered normal for rats of this age and strain. These included slightly higher levels of total protein, and total globulin values for males at 300 mg/kg, slightly higher levels of creatinine for males at 100 and 300 mg/kg, a slightly lower albumin/globulin (Alb/Gl) ratio for females at 1000 mg/kg, a slightly higher glucose level for females at 300 and 1000 mg/kg, slightly higher potassium and chloride levels for females at 100, 300 (not statistically significant) and 1000 mg/kg, and a slightly lower calcium concentration for females at 300 mg/kg.

NEUROBEHAVIOUR
Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all selected animals. The variation in motor activity did not indicate a relation with treatment. All groups showed a similar habituation profile with very high activity in the first interval that decreased over the duration of the test period.

ORGAN WEIGHTS
The following (statistically significant) changes in organ weights were noted:
− Higher liver weight for females at 300 mg/kg (absolute; 113% of control) and 1000 mg/kg (absolute and relative; 116% of control).
− Higher liver weight for males at 1000 mg/kg (relative to body weight only; 110% of control).
− Lower thymus weight (absolute: 66% of control and relative: 74% of control) for males at 1000 mg/kg.
− Higher adrenal weight for females at 1000 mg/kg (absolute: 122% of control and relative: 125% of control).
− Higher spleen weight for females at 1000 mg/kg (absolute: 127% of control and relative: 129% of control).
Any other statistically significant changes were considered to be of no toxicological significance as they were due to the lower body weight noted for males, occurred in the absence of a treatment related distribution, remained within the range considered normal for rats of this age and strain and/or no microscopic correlate was found. These findings included a higher relative brain weight and lower absolute weight of seminal vesicles for males at 1000 mg/kg, and higher absolute thymus weight for females at 300 mg/kg.

GROSS PATHOLOGY
There were treatment-related macroscopic findings in female rats treated at 1000 mg/kg consisting of irregular forestomach epithelium in 2/10 animals and thickened wall of the small intestines in 3/10 animals. A yellowish reddish soft nodule (size 31x26 mm) was noted in the skin/subcutis of the chest region of a male treated at 300 mg/kg. The finding consisted of and was microscopically correlated to a carcinoma of the subcutis. Because of the low incidence, early occurrence and absence of proliferative lesions in the skin/subcutis of other treated rats in this study, this was considered to be an incidental finding, unrelated to treatment. All remaining findings noted for both treated and control animals, occurred at an incidence that was within background levels and/or in the absence of any dose-related distribution. Therefore, they were not considered to be toxicologically relevant.

HISTOPATHOLOGY: NON-NEOPLASTIC
Adverse histopathological alterations were present in the mesenterial lymph nodes of males and females treated at 300 and 1000 mg/kg, and consisted of foamy macrophage foci and granulomatous inflammation (with or without necrosis). These foamy macrophage foci were considered to be related to intestinal uptake of the test article starting in the villi of the small intestines (recorded as foamy macrophage in the villi of the small intestines in rats treated at 300 mg/kg and 1000 mg/kg; macroscopic correlate: thickened wall of the small intestine) and from there transported to the mesenteric lymph node. When taking up more material at higher doses (300 and 1000 mg/kg), the foci in the mesenteric lymph node increased in incidence and/or severity. Additionally, development into granulomas (sometimes with central necrosis, up to marked degree) were recorded in rats treated at 300 and 1000 mg/kg. The presence of foamy macrophage foci up to slight degree in the mesenteric lymph node in the majority of rats treated at 100 mg/kg without development of granulomas was considered non-adverse in nature. Other treatment-related morphologic findings were recorded in the lung (alveolar macrophages) of males and females at 300 and 1000 mg/kg, liver (hepatocellular centrilobular hypertrophy; corresponding increased organ weight) of males and females at 1000 mg/kg, spleen (decrease in hematopoietic foci) of males at 1000 mg/kg, and adrenals (hypertrophy of the zona fasciculate with corresponding increased adrenal weights) of females at 1000 mg/kg. Also local effects on the forestomach (diffuse hyperplasia of the squamous epithelium; correlating macroscopic finding: irregular epithelium) were noted for males and females at 1000 mg/kg. Additional microscopic findings in one female (no. 79) at 1000 mg/kg killed in extremis on Day 8 of the mating period consisted of moderate lymphoid atrophy in the thymus and moderate bone marrow atrophy.

Effect levels

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

1. Females 58 and 59 were interchanged at start of mating. Consequently, the clinical signs, body weights and food consumption data and the mating date recorded for animal no. 58 during the mating period actually belongs to female 59, and vice versa. Upon discovery of the interchange, females 58 and 59 were reallocated as 81 and 82, respectively. All subsequent data were collected as animals 81 and 82. Additionally, these animals were not re-tattooed with the new animal number, though they had their original permanent identification.

Evaluation: Since both animals were within the same treatment group, summary data were unaffected by this interchange. The animals still had their original tattoo and the new numbers were clearly documented on the cage cards.

2. Animal no. 55 (300 mg/kg) was not weighed in the tox program as scheduled on 03-Jan-2013.

She was weighed a few days later, on 07-Jan-2013. Evaluation: The dosing volume (1.3 mL) remained the same for this animal, so there was no impact on dosing and the few days difference between body weight collection has no impact.

3. No body weight was collected for female no. 44 on lactation Day 4 and no food consumption data are available from lactation Days 1-4.

Evaluation: Sufficient data is available for a thorough evaluation.

4 On Day 4 no observations were entered online for pup 2 from litter 44 (control), and no observations were entered online on Day 6 for all pups from litter 66 (300 mg/kg).

Evaluation: Sufficient data is available for a thorough evaluation, and no clinical signs were

noted for pups in the treated groups during the study.

5. The organs and tissues from animal no. 52 were fixed in Modified Davidson’s solution instead of formalin.

Evaluation: Only a skin abnormality was required for microscopic examination; this was unaffected by the difference in fixatives.

The study integrity was not adversely affected by the deviations.