2,3-epoxypropyltrimethylammonium chloride

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: OECD 407

Type of information:

other: Assessment report

Adequacy of study:

other information

Study period:

2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Although the EU risk assessment report is secondary literature, all data and risk assessment for the human, health and the environment have been evaluated and reviewed by Finland prior to publication. The risk assessment report has been submitted to final approval and published in the Official Journal of the European Union C157/10 dated on 21.06.2008. Thus, it is considered the information reported are reliable with the restrictions that reliability of the data presented has not been assessed again.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

European Union Risk Assessment 2,3-epoxypropyltrimethylammonium chloride CAS RN 3033-77-0 Einecs No: 221-221-0

Author:

EC

Year:

2008

Bibliographic source:

Risk Assessment. Final approved version. Rapporteur: Finland (FIN). European communities. Printed in Italy. 147pp

Materials and methods

Principles of method if other than guideline:

There are no studies conducted on toxicity to fertility. Some information of the effect on the morphology of the gonads can be obtained from the 28-day test conducted with Wistar rats (see 3033-77-Repeated dose toxicity: oral/2008/EURA,ss).

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,3-epoxypropyltrimethylammonium chloride

EC Number:

221-221-0

EC Name:

2,3-epoxypropyltrimethylammonium chloride

Cas Number:

3033-77-0

Molecular formula:

C6H14NO.Cl

IUPAC Name:

N,N,N-trimethyl(oxiran-2-yl)methanaminium chloride

Details on test material:

- Physical state: aqueous solution
- Analytical purity: 72.6 % test substance in aqueous solution
- Impurities (identity and concentrations): The original solution had about 10% impurities

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

No data

Details on mating procedure:

Not applicable.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

28-days.

Frequency of treatment:

1 time daily/7days/week

Details on study schedule:

No data

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 3.16, 10.0, 31.6 or 100 mg/kg EPTAC (72.6%) bw/day
Basis:
actual ingested

No. of animals per sex per dose:

5 animals/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

No data

Examinations

Parental animals: Observations and examinations:

Yes. See "3033 -77 -0 Repeated dose toxicity: oral/2008/EURA" for a more complete description of the study.

Oestrous cyclicity (parental animals):

Not applicable (OECD 407)

Sperm parameters (parental animals):

Not applicable (OECD 407)

Litter observations:

Not applicable (OECD 407)

Postmortem examinations (parental animals):

Yes. See "3033 -77 -0 Repeated dose toxicity: oral/2008/EURA" for a more complete description of the study.

Postmortem examinations (offspring):

Not applicable (OECD 407)

Statistics:

Yes but not detailed.

Reproductive indices:

Not applicable (OECD 407)

Offspring viability indices:

Not applicable (OECD 407)

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

effects observed, treatment-related

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Details on results (P0)

CLINICAL SIGNS AND MORTALITY
Most high dose animals exhibited in their clinical picture piloerection and sunken sides. Two females and one male showed disturbed general
condition before death or decreased muscle tone. In the high dose group, four females and one male died during the last week of administration. One female was sacrificed moribund at day 16 of treatment

BODY WEIGHT AND WEIGHT GAIN
In the 31.6 and 100 mg/kg groups, the bodyweight development slowed in parallel with the changes in food consumption. The body weight gain in
the 31.6 and 100 mg/kg dose groups was significantly reduced during weeks 2-4 of the treatment period. The body weight in the 100 mg/kg group males at day 28 was about 45% lower than the control group and in the 31.6 mg/kg group at that time the mean weight was about 18% lower and in
the 10 mg/kg group a 12 % reduction was noted. The 100 mg/kg dose females had about 38 % lower mean body weight than the control.
No significant difference was noted in other groups or in the recovery group. After the recovery period, the male 100 mg/kg recovery group had
still about 29% lower mean body weight, when compared to control.

FOOD CONSUMPTION AND COMPOUND INTAKE
The food consumption decreased in the mid- and high-dose groups.
The food consumption dropped to less than 50 % of the controls within two weeks in high dose females and within four weeks in high dose males.
Unlike the female rats, who had an almost complete recovery in their food consumption one week after the end of administration, the male recovery
returned to normal after seven weeks.

ORGAN WEIGHTS
In organ weight measurements, male testis (-35-40%) and brain weight (-10 %) were reduced in the high dose group and remained so in the recovery group at week 9. Also the brain to body weight ratio and brain to testis weight ratio were bigger in that group. As stated above, the body weight in
the high dose group was about 50% less than that of the control animals. The male absolute liver weight was slightly reduced in the 10 (19 %) mg/kg and 100 mg/kg (50 %) group with no change in the ratio to body weight. The only significant organ weight change that correlated with the
administrated dose was found in the heart (Males from low to high dose: -21 %, -26 %, 43 %). In females, the absolute weighs of liver (41 %) and heart (-38%) were decreased in the high dose group without a change in the ratio to body. Ovary weights were reduced in the 31.6 (~40 %) and 100 mg/kg (max 60 %) groups.

GROSS PATHOLOGY
Gross macroscopical examination revealed small spleen and thymus in both sexes of the high dose group than in other groups, which in microscopy was seen as a reduction of lymphatic tissue. The effect on thymus could not be adequately assessed in the intermediate dose groups,
as this organ was not taken at necropsy. The uterus of the two highest doses was reduced in size. However, the spleen, thymus or uterus weights
were not reported.

HISTOPATHOLOGY:
In the microscopic observations, the proximal convoluted tubule (PCT) of the kidneys had dose related necrosis and vacuolisation (lipid or fat deposits). In the 3.16 mg/kg dose group 3/5 males had minimal and 2/5 slight vacuolisation. All female rats of the 3.16 mg/kg group had minimal
vacuolisation. In the 10 mg/kg dose group, slight vacuolisation was present also in females (3/5). There was one case of minimal necrosis and hyperplasia, both in male rats of the 3.16 mg/kg group. One control male also showed tubular hyperplasia. In the 10 mg/kg group, proximal tubular
vacuolisation was minimal in 5/10 and slight in 5/10 animals, minimal necrosis and hyperplasia were present in 7/10 animals. Nuclear polyploidy
was present in 7/10 rats and 6/10 of those also had hyperplasia. The 31.6 mg/kg 7/10 animals of both sexes showed slight to 3/10 moderate
vacuolisation of the kidney proximal tubular cells, proximal tubular hyperplasia (minimal in 8/10, slight in 2/10), and polyploid nuclei in the proximal tubules and one case of an abnormal mitotic figure and reduced cellularity of the bone marrow. Minimal PCT-necrosis was present 9/10 animals
of this group. Minimal testicular atrophy was observed in 1/5 males and follicular atrophy of the ovaries was observed in 5/5 females (1/5 slight, 2/5 moderate, 4/5 marked). Persistent corpora lutea were reported in 4/5 females (2/5 moderate, 2/5 marked).
In the high dose group, vacuolisation was seen the proximal tubular cells of the kidney of inall animals and graded slight (1/10), moderate (5/10), marked (3/10) or massive (1/10). In the recovery group, the changes were slight in 5/10, moderate in 5/10 animals. Minimal PCT hyperplasia was
present in 6/10 animals of the dose group, two of the remaining animals had moderate to marked hyperplasia of the collective tubules and
transepithelial cells. In the recovery group PCT hyperplasia (minimal to moderate) was observed in 8/10 animals. Also minimal to moderate nuclear polyploidy was seen in 9/10 dose group and 9/10 recovery group animals. Minimal to moderate necrosis was present in the PCT epithelium of all
animals which was noted in the papilla of one male rat as marked.
Testicular atrophy was minimal in 1/5 males, moderate in 1/5 males and marked in 1/5 males of the dose group and minimal, slight or moderate in 1each of 5 recovery group males. Follicular atrophy of the ovaries was moderate in 2/5, marked in 1/5 and massive in 1/5 females of the dose group and slight in 1/5, marked in 3/5, and massive in 1/5 animals of the recovery group. Persistent corpora lutea were reported in 4/5 females of the
dose group (3/5 marked, 1/5 moderate) and 5/5 animals of the recovery group. Two of the recovery group females died after 14 and 16 days of
treatment.
In the spleen lymphoid atrophy (minimal 1/5, slight 1/5, moderate 1/5, marked 1/5) was observed in 5/5 males and 4/5 females (1/5 minimal, 1/5
slight, 2/5 marked) of the dose group and the 2/5 females that died premature in the recovery group. None of the animals that
survived the recovery period had lymphoid atrophy in the spleen.
The changes in the kidneys and the gonads persisted after the 4-week recovery period (100 mg/kg).
Again, only in the two highest doses, there was a reduction of all cell lines in the bone marrow, which correlated with the white blood cell reduction
seen in the haematology. The high dose animals showed occasional maturation disorders, vacuolisation and abnormal mitoses in the bone marrow
cells. The changes in the bone marrow, thymus or spleen were reversible after the recovery period.
Some animals of the high dose animals had mild focal hyper- and parakeratosis of the forestomach or mild erosions and haemorrhages in the
glandular stomach attributed to the local irritant property of EPTAC. The small intestine of these animals had villous atrophy and crypt necrosis. The changes in the stomach and small intestine were reversible.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

10 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: based on the severe morphological changes in the reproductive organs of both sexes (observed in repeated dose toxicity, OECD 407)

Results: F1 generation

Details on results (F1)

Not applicable.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

No other information was available.

Applicant's summary and conclusion

Conclusions:

The 10 mg/kg NOAEL obtained from the 28-day repeated dose toxicity study is selected for toxicity to reproduction. EPTAC is therefore classified as Repr.2, H361f (Suspected of damaging fertility) according to the criteria of the CLP regulation (No 1272/2008) and as Repr. Cat. 3; R62 (Possible risk of impaired fertility) according to the criteria Directive 67/548/EEC.

Executive summary:

There are no studies conducted on toxicity to fertility. Some information of the effect on the morphology of the gonads can be obtained from the 28-day test conducted with Wistar rats. The rats were administered by oral gavage 0, 3.16, 10.0, 31.6 or 100 mg/kg EPTAC (72.6 %) for 28 days (Degussa, 1990). The control and high dose group had 10 males and 10 female and the low- and mid-dose groups had five rats of each sex. Five high dose and five control group animals were submitted to a 4-week post-exposure observation period. The study was conducted under GLP regulations and OECD guideline 407 was followed as the study protocol. See section 7.5.1 "3033 -77 -0 Repeated dose toxicity: oral/2008/EURA" for a more complete description of the study.

Male rats of the highest dose groups had a significant decrease (-35-40%) of the testis weight at week 5, which remained significantly lower in the recovery group at week 9. The female ovaries showed a weight decrease trend at week 5, where the weight difference was significant in the dose groups 31.6 mg/kg and 100 mg/kg. However, at week 9 in the recovery group, there was significant ovary weight decrease (max 60%) in the right ovary only in the 100 mg/kg dose group. Microscopically, males of the 31.6 and 100 mg/dose groups showed a dose-related incidence and severity of focal atrophy of the testes. Females had a dose dependent severity of follicular atrophy and persistent corpora lutea in the 31.6 and 100 mg/kg dose groups. The uterus epithelium was atrophic and morphologically in anoestrus. The changes in the gonads persisted in both sexes through the recovery period. The body weight in the 100 mg/kg group males at day 28 was about 45% lower than the control group and in the 31.6 mg/kg group at that time the mean body weight was about 18% lower and in the 10 mg/kg group a 12 % reduction was noted. The 100 mg/kg dose females had about 38 % lower mean body weight than the control. No significant difference was noted in other groups or in the recovery group. After the recovery period, the male 100 mg/kg recovery group had still about 29% lower mean body weight, when compared to control.

Although these results tell little of the effect on the reproductive performance itself they can be used to set an indicative NOAEL based on the rather severe morphological changes in the reproductive organs of both sexes. The 10 mg/kg NOAEL obtained from the 28-day repeated dose toxicity study is selected for toxicity to reproduction. EPTAC is therefore classified as Repr.2, H361f (Suspected of damaging fertility) according to the criteria of the CLP regulation (No 1272/2008) and as Repr. Cat. 3; R62 (Possible risk of impaired fertility) according to the criteria of the Directive 67/548/EEC.