N-butyl-2,2,6,6-tetramethylpiperidin-4-amine, oligomeric reaction products with N,N'-bis(3-aminopropyl)ethylenediamine and 2,4,6-trichloro-1,3,5-triazine

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

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:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: FDA guideline for testing food addtives (Toxicological Principles for the Safety Assessment of Direct Food Additives and Colour additives used in food; US Food and Drug Administration Bureau of Foods, 1982)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-butyl-2,2,6,6-tetramethylpiperidin-4-amine, oligomeric reaction products with N,N'-bis(3-aminopropyl)ethylenediamine and 2,4,6-trichloro-1,3,5-triazine

EC Number:

500-311-6

EC Name:

N-butyl-2,2,6,6-tetramethylpiperidin-4-amine, oligomeric reaction products with N,N'-bis(3-aminopropyl)ethylenediamine and 2,4,6-trichloro-1,3,5-triazine

Cas Number:

120498-03-5

IUPAC Name:

1,3-Propanediamine,N-N’’-1,2-ethanediylbis- reaction product with 1,3,5-triazine-2,4-diamine,N,N’-dibutyl-6-chloro-N,N’-bis(2,2,6,6-tetramethyl-4-piperidinyl), polymer with N-butyl-4,6-dichloro-N-(2,2,6,6-tetramethyl-4-piperidinyl)-1,3,5-triazin-2-amine

Details on test material:

Purity 98% min.
Appearance: white ivory powder
Storage conditions: room temperature
Expiry date: July 1998

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Young make and female Sprgue Dawley rats obtained from a colony maintained under SPF- conditions at Charles River, Germany. The animals, 90 males and 90 females, arrived on the 5 February 1997 when they were about 3 weeks old. Upon arrival the rats were checked for signs of ill health and anomalies and kept in quarantine. During the quarantine period, the microbiological staus of the rats was assessed, these results were satisfactory. The body weights on the experimental start date ranged from 125.8 g to 172.4 g for males and 105.7 g to 138.0 g for females.
Environmental conditions: The air in room was changed approx. 10 times per hour. They received 12 hours o flight and 12 hours of dark. The temperature in teh room was maintained between 20 and 24 °C (day 88 tp day 91 the temperature went up to a maximum of 25.5°C). The relative humidity was between 30 and 55%. The rats were housed in groups of 5 according to their sex.

Administration / exposure

Route of administration:

oral: feed

Details on oral exposure:

The test substance was incorporated into the feed. In order to obtain constant dose levels in terms of the animals body weight, the dietary levels of the test substance were adapted weekly. For this purpose fresh batches of test diets were prepared weekly and given to animals on the day of preparation. Halfway each week the feed in the feeders was replaced by a fresh protion from the freezer (<-10°C).

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability of the test substance under simulated experimental conditions was investigated by analyzing diet samples in teh concentration range used in the study on the day of preparation, after storage for 2 weeks in a freezer. From teh first batch of diets prepared for the 13-week study, five samples of each of the three concentration levels, taken at different locations in teh feed container, were analysed to confirm the homogeneity and content of the test substance in teh diets. In addition, the content of the test substance was checked by analysis in two other batches of diets (those prepared for weeks 5 and 10).

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily- assessed weekly

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
Group 1879 A
Basis:
other: 0 mg/ kg body weight/ day

Doses / concentrations 2

Remarks:

Doses / Concentrations:
Group 1879 B
Basis:
other: 5 mg/ kg body weight/ day

Doses / concentrations 3

Remarks:

Doses / Concentrations:
Group 1879 C
Basis:
other: 13 mg/ kg body weight/ day

Doses / concentrations 4

Remarks:

Doses / Concentrations:
Group 1879 D
Basis:
other: 24 mg/ kg body weight/ day

No. of animals per sex per dose:

20 females and 20 males per dose group

Control animals:

yes, plain diet

Details on study design:

The study comprised four groups of 20 rats/ sex each, viz. one control group (A) and three treatment groups (B, C and D) receiving different levels of the test substance. Treatment was started on 18 February 1997 (nominal day 0). The study was terminated with the autopsy of the male rats on 20 and 21 May 1997 (nominal days 91 and 92) and of th efemale rats on 22 and 23 May 1997 (nominal days 93 and 94).

Examinations

Observations and examinations performed and frequency:

Clinical signs: Each animal was removed from its cage and examined physically twice prior to initiation and once a week during the treatment period (this examination coincided with the recording of body weights). In addition they were observed daily in th emorning by cage side observations and if necessary handled to detect signs of toxicity. On working days all cages were checked in the afternoon for dead or moribund animals to minimise loss. All abnormalities were recorded.
Body weights: The body weight of each animal was recorded during the pretreatment period (day -7); when starting the administration of the test substance (day 0), and once weekly thereafter. In addition the animals were weighed on the scheduled sacrifice day in order to calculate their organ to body weight ratios. Body weights were also recorded two days after arrival to monitor adequate growth during acclimatisation period.
Food conversion and consumption: Food consumption was measuered starting on teh day of allocation (ie. one week before initiation of treatment), by weighing th efeeders. The results were expressed as gram per rat per day. The consumption was measured over periods of 7 days or 6 days. The efficiency of the food utilisation was calculated and expressed in gram weight gain per gram food consumed.

Sacrifice and pathology:

In week 14 all animals were killed on a number of successive working days (males on day 91 and 92; females on 93 and 94) in such a sequence that the average time of killing was approximately the same for each group. The animals were killed after overnight fasting by exsanguination from the abdominal aorta under ether anaesthesia. Subsequently they were examined macroscopically for pathological changes.
The following organs were weighed (paired organs together) as soon as possible after dissection to avoid drying: adrenals, brain, epididymides, kidneys, liver, lungs, testes.
Samples of the following tissues of all animals were preserved in a neutral aqueous phosphate-buffered 4 per cent solution of formaldehyde (the eyes were fixed in glutaraldehyde/paraformaldehyde and later preserved in formalin: the lungs and urinary bladder were infused with formalin to insure fixation): adrenals, axillary lymph nodes, brain (brain stem, cerebaum, cerebellum), caecum, colon, epididymides, eyes (with optic nerve), heart, kidney, liver, lungs, mandibular lymph nodes, mediastinal lymph nodes, mesenteric lymph nodes, nasopharynx, oesphagus, ovaries, pancreas, pituitary, rectum, small intestine, spinal cord, spleen, sternum, stomach, salivary glands, testes, thymus, thyroid with parathyroids, trachea, urinary bladder, uterus, all gross lesions. In addition as much fat tissue as possible was collected from the abdominal cativity of all animals at autopsy. the collected fat tissue was weight, frozen in liquid nitrogen and stored in the freezer <-10°C to enable possible future chemical analysis of the test substance in fat tissue.

Other examinations:

Haematology: At autopsy blood was collected from 10 rats/ sex/ group whilst under ether anaesthesia. The animals were fasted overnight prior to blood colelction. Blood was collected from teh abdominal aorta in tubes containing K2-EDTA as anticoagulant. The following measures were conducted: haemoglobin, packed cell volume, redblood cell count, reticulocytes, total white blood cell count, differential white blood cell count, prothrombin time, activated partial thromboplastin time, thrombocyte count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpusculr haemoglobin concentration.

Statistics:

The statistical procedures used to evaluate the results are as follows:
Body weights: one-way analysis of convariance using pre-exposure (day 0) weights as the covariate. When group means were significantly different (p<0.05), individual pairwise comparisons were made using Dunnett's multiple comparison method;
Food consumption, food conversion efficiency, red blood cell and coagulation variables, total white blood cell counts, absolute differntial white blood cell counts, climical chemistry and organ weights: analysis of variance (Anova) followed by Dunnett's multiple comparison tests;
Precentages of diffential white blood cell counts: Krushkal-Wallis nonparametric one-way analysis of variance. When teh analysis yielded a significant difference, pairwise comparisons between teh control- and th etreatment groups were made by means of Mann-Whitney U-tests:
Histopathological changes: Fisher's exact probabiltiy test.
All analyses were two-sided. Group mean differences with an associated probability of less than 0.05 were considered to be statistically significant.

Results and discussion

Results of examinations

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):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

The dietary administraion of UVASORB HA88 for 13 weeks to rats did not cause any changes nasopharynx, lungs, mediastinal lumph nodes, liver and meenteric lymph nodes. No test substance particles were encountered in any organ examined. Also, in none of these organs foreign body reaction was seen.

Effect levels

1

Dose descriptor:

NOEL

Remarks:

Mortality

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

mortality

2

Dose descriptor:

NOEL

Remarks:

Clinical signs

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

3

Dose descriptor:

NOEL

Remarks:

Body weights

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

4

Dose descriptor:

NOEL

Remarks:

Haematology

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

haematology

5

Dose descriptor:

NOEL

Remarks:

Clinical chemistry

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical biochemistry

6

Dose descriptor:

NOEL

Remarks:

Organ weights

Effect level:

24 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

organ weights and organ / body weight ratios

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The results obtained in the present sub-chronic study did not reveal any adverse affects of UVASORB HA88 in rats.
Therefore it is concluded that the feeding of UVASORB HA88 in the diet at levels providing an intake up to 24mg/kg body weight/ day for 13 weeks did not result in any signs of toxicity