Reaction mass of Ethanaminium, 2-hydroxy-N,N-dimethyl-N-[2-[(1-oxooctadecyl)oxy]ethyl]-, chloride and Ethanaminium, N,N-dimethyl-2-[(1-oxohexadecyl)oxy]-N-[2-[(1-oxooctadecyl)oxy]ethyl]-, chloride

Toxicity to reproduction

Administrative data

Endpoint:

toxicity to reproduction

Remarks:

other: subacute repeated dose toxicity study in rodents

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-02-09 to 2009-03-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral)) 30 May 2008)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 407, adopted 03 October 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

MDEA Esterquat C18 satd.

IUPAC Name:

MDEA Esterquat C18 satd.

Details on test material:

- Name of test material (as cited in study report): Dimethylbis[2-[(1-oxooctadecyl)oxy]ethyl]ammonium chloride
- Physical state: solid
-Analytical purity: 98.5%

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 8 weeks
- Weight at study initiation: Females: 187 - 223 g (mean: 201 g); Males: 241 - 290 g (mean: 266 g)
- Housing: Full-barrier in an air conditioned room. The animals were kept individually in IVC cages.
- Diet (e.g. ad libitum): ad libitum (Altromin 1324 maintenance diet)
- Water (e.g. ad libitum): ad libitum (sulphur acidified tap water)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2009-02-09 To: 2009-03-30

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

PREPARATION
The test item formulations were prepared freshly on each administration day, before the administration procedure.
The test item was dissolved while stirring and heated up to approx. 70° C in Aqua ad injectionem (sterile water). As guaranteed by the sponsor this procedure did no affect the chemical stability of the test compound.

VEHICLE
- Concentration in vehicle: 10% w/w
- Amount of vehicle (if gavage): 10 mL/kg bw
- Lot/batch no. (if required): Lot 7494A191, expiry date: Nov. 2010
- Purity: aqua ad injectionem

Details on mating procedure:

animals were not mated

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis of the dosing preparations of the test item in the vehicle (nominal concentration) was performed once a week by two-phase titration.

Duration of treatment / exposure:

28 days

Frequency of treatment:

once daily, 7 days per week

Doses / concentrations

Remarks:

Doses / Concentrations: 62.5, 250, 1000 mg/kg bw/d; Basis: actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control:

No

Examinations

Oestrous cyclicity (parental animals):

not examined

Sperm parameters (parental animals):

not examined

Litter observations:

n.a.

Postmortem examinations (offspring):

n.a.

Reproductive indices:

n.a.

Offspring viability indices:

n.a.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Effect levels (P0)

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: No significant treatment related effects on reproductive organs (weights and histopathology) were seen at the highest dose administered

Results: P1 (second parental generation)

Effect levels (P1)

Remarks on result:

other: No P1 generation was used in this 28 day sub acute study.

Results: F1 generation

Effect levels (F1)

Remarks on result:

other: No F1/F2 offspring generations were used in this subacute 28 day study.

Results: F2 generation

Effect levels (F2)

Remarks on result:

other: No F1/F2 offspring generations were used in this subacute 28 day study.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

ORGAN WEIGHTS

There were no significant differences in relative and absolute organ weights for both sexes and any of the groups except prostata weight which was absolutely and relatively higher in the HD recovery group as compared to the control group values. In absence of finding in the main study animals the toxicological relevance of this finding is difficult to assess and it may be consider as incidental, moreover as the variation is below 20%.

GROSS PATHOLOGY

There were no test substance-related macroscopic observations in either male or female rats.

HISTOPATHOLOGY

Histopathological changes seen at terminal sacrifice were few and all of them were considered to be incidental in origin and/or within the range of expected changes for rats of this age and strain kept under laboratory conditions. There was no indication of any test item-related effect on organs and tissues evaluated in this study.

Applicant's summary and conclusion

Conclusions:

The results from the evaluation of reproductive organs (organ weights of ovary, uterus with cervix, testis, epididymides, prostate with seminal vesicles and coagulating glands and the gross pathology and the histopathology of the reproductive organs (gonads, prostate and seminal vesicle, epididymides, uterus and vagina)) revealed no test substance related findings in the main study up to and including the highest tested dose of 1000 mg/kg bw/day.

Executive summary:

In this Repeated Dose, 28-day Oral Toxicity Study according to OECD Guideline 407 (adopted 03 October 2008) and EU method B.7 (30 May 2008), the test item MDEA Esterquat C18 satd. suspended in Aqua ad injectionem (10 % w/w) as vehicle was orally administered in graduated doses to three groups of male and female rats (HsdRccHan: WIST) by gavage.

The main study included 4 groups (control, 62.5, 250, 1000 mg/kg bw) with each 5 male and 5 female animals and the recovery study included 2 groups (control, 1000 mg/kg bw) each with 5 male and 5 female animals.

All animals but one (male; 250 mg/kg bw group) treated with the test item MDEA Esterquat C18 satd. survived throughout the test period and were sacrificed on day 29 or day 43 for the recovery animals. The death of animal No. 25 was not test-item related and a gavage accident may have been the cause of death of this animal.

No test substance-related findings were detected or observed in clinical examinations, body weights, food consumption values, haematology, urinalysis, neurobehavior, clinical biochemistry or gross pathology evaluations, including organ weights of ovary, uterus with cervix, testis, epididymides, prostate with seminal vesicles and coagulating glands and the gross pathology and the histopathology of the reproductive organs (gonads, prostate and seminal vesicle, epididymides, uterus and vagina).

  

The NOEL is 1000 mg/kg bw/day in this study.