Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: LD50 > 2000 mg/kg bw, rat, OECD Guideline 423, GLP compliant

Acute dermal toxicity: LD50 > 2000 mg/kg bw, rat/rabbit, OECD Guideline 402, GLP compliant

Acute inhalation toxicity: inhalation is no relevant route of exposure

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, GLP
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
(30 May 2008)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
(adopted 17 December 2001)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: Wistar Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: approx. 10 weeks
- Weight at study initiation:
- Fasting period before study: yes, overnight prior to dosing and until approximately 1 hour after the second administration
- Housing: in groups of 3
- Diet (e.g. ad libitum): pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany); ad libitum
- Water (e.g. ad libitum): tap water; ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40-70%
- Air changes (per hr): 15/h
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage): 2x10 mL/kg bw within 24 h
- Justification for choice of vehicle: selected based on trial formulations performed at the testing laboratory

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg bw

DOSAGE PREPARATION (if unusual):
Formulations (w/w) were prepared within 5 hours prior to dosing and were homogenized to visually acceptable levels.
The formulations were released from the formulation unit in a water bath containing water of 35±5°C

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines.
Doses:
2000 mg/kg bw , administered as two dosages of 1000 mg/kg bw within 24 hours (first at t=0 and second at t=4 hours)
No. of animals per sex per dose:
6 females (each dose group consisted of 3 animals)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: mortality: twice daily; Clinical signs: at periodic intervals on the day of dosing (day 1) and once daily thereafter; weighing: days 1 (pre-administration), 8 and 15
- Necropsy of survivors performed: yes
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: No mortality was observed
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture was noted for all animals (with piloerection for one animal) on days 1 and 2.
Body weight:
The body weight gain shown by all animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 of MDIPA-Esterquat C16-18 and C18 unsatd. in female rats was >2000 mg/kg bw.
Executive summary:

In an acute oral toxicity study according to OECD guideline 423 (17 December 2001) and EU method B.1 tris (30 May 2008), 6 fasted, approx. 10 weeks old female Wistar Crl:WI (Han) rats were given an oral dose of MDIPA-Esterquat C16-18 and C18 unsatd. (100% a.i.) at a limit dose of 2000 mg/kg bw administered as two dosages of 1000 mg/kg bw within 4 h. Animals were then observed for 14 days.

No animal died during the observation period, hunched posture was noted for all animals (with piloerection for one animal) on days 1 and 2.

The body weight development of the animals was within normal range for animals of the same age and strain. The necropsy at the end of the observation period showed no macroscopically visible test substance related pathologic organ findings.

Oral LD50 Females > 2000 mg/kg bw

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute oral toxicity

In an acute oral toxicity study according to OECD guideline 423 (17 December 2001) and EU method B.1 tris (30 May 2008), 6 fasted, approx. 10 weeks old female Wistar Crl:WI (Han) rats were given an oral dose of MDIPA-Esterquat C16-18 and C18 unsatd. (100% a.i.) at a limit dose of 2000 mg/kg bw administered as two dosages of 1000 mg/kg bw within 4 h. Animals were then observed for 14 days. No animal died during the observation period, hunched posture was noted for all animals (with piloerection for one animal) on days 1 and 2. The body weight development of the animals was within normal range for animals of the same age and strain. The necropsy at the end of the observation period showed no macroscopically visible test substance related pathologic organ findings. The oral LD50 for female rats was > 2000 mg/kg bw.

 

Also the two source substances used for read-across for the endpoints mutagenicity, repeated dose toxicity and toxicity to reproduction , MDIPA-Esterquat C18 unsatd. and MDEA-Esterquat C16-18 and C18 unsatd., were of equally low oral toxicity as demonstrated by the following studies on rats and mice:

 

In an acute oral toxicity study according to OECD guideline 423 (17 December 2001) and EU method B.1 tris (30 May 2008), 6 fasted, approx. 11 weeks old female White Wistar, HsdCpb: WU rats were given a single oral dose of undiluted MDIPA-Esterquat C18 unsatd. (100% a.i.) at a limit dose of 2000 mg/kg bw and observed for 14 days. No animal died during the observation period, no clinical signs were observed. The body weight development of the animals was positive and within normal range. The necropsy at the end of the observation period showed no macroscopically visible test substance related pathologic organ findings. The oral LD50 for female rats was > 2000 mg/kg bw.

 

This finding was supported by an acute oral toxicity study similar to OECD guideline 401, with mice. Fasted, 20±2 g mice of Tylers Original Strain (5/sex) were given a single oral dose of MDIPA Esterquat C18 unsatd. (75% a.i.) at a dose of 6.6 mL/kg bw (corresponding to 5.0 mL/kg bw adjusted to purity) and observed for 14 days. 1/10 animals died on day 4. Slight signs of toxicity were observed in the high dose group. However, all animals appeared asymptomatic 24 h after administration.

The oral LD50 for male and female mice was > 5000 mL/kg bw (as a.i.). Based on a density of 0.987 g/cm³ this corresponds to an LD50 of > 4935 mg a.i./kg bw.

 

Two studies for acute oral toxicity are available for MDEA-Esterquat C16-18 and C18 unsatd., one limit test in rats and an Irwin dose-range study in mice.

 

In an acute oral toxicity study comparable to OECD Guideline 401 (24. Feb. 1987, now deleted) rats were dosed once with 10 000 mg/kg bw MDEA-Esterquat C16-18 and C18 unsatd. and observed for 14 days. No mortality occurred in this limit test. There were no treatment related clinical signs, necropsy findings or changes in the body weight.

 

The second study with mice was performed as an Irwin dose-range finding study with oral application of 316, 1000, 3160 and 10 000 mg/kg bw of MDEA-Esterquat C16-18 and C18 unsatd. and with an observation period of 7 days. All mice dosed with 10 000 mg /kg bw showed slight apathy between 10 and 20 minutes after dosing. In addition, one animal in this group showed further indication of CNS depression when observed at 30 and 90 minutes after treatment. This animal appeared normal during the observations performed at 150 and 300 minutes; however it died overnight between day one and day two. No other treatment related clinical signs, were recorded in the remaining animals at any time during the study. Necropsy was not performed after study termination; changes in body weights were not recorded. MDEA-Esterquat C16-18 and C18 unsatd. is practically non-toxic based on the oral LD50 of > 10 000 mg/kg bw obtained in the two studies.

 

Acute dermal toxicity

Testing of acute dermal toxicity of MDIPA-Esterquat C16-18 and C18 unsatd. is scientifically not justified as reliable and relevant data from two structurally related substances are used to fulfil this information requirement in a weight of evidence approach.

 

In an acute dermal toxicity study according to OECD guideline 402 (24 February 1987) and EU method B.3 (30 May 2008), groups of young adult White Wistar, HsdCpb: WU rats (5/sex) were dermally exposed to the source substance MDIPA Esterquat C18 unsatd. (50% in sesame oil) for 24 hours to approx. 10% body surface area at a limit dose of 2000 mg/kg bw.  Animals then were observed for 14 days. No animal died during the observation period. No symptoms of systemic toxicity were observed. The body weight development of the animals was positive 7 and 14 days after application. However, body weight gain was slightly lower than expected. The necropsy of all animals 14 days after application showed no macroscopically visible test substance related pathologic organ findings. The dermal LD50 for male and female rats was > 2000 mg/kg bw.

 

Also the second source substance used for read-across MDEA-Esterquat C16-18 and C18 unsatd. was of low dermal toxicity:

In an acute dermal toxicity study (comparable to OECD guideline 402), groups of 3 male and female New Zealand White rabbits were dermally exposed to MDEA-Esterquat C16-18 and C18 unsatd. (40 % a.i) in oleum arachidis for 24 hours to10 x 15 cm of body surface area at doses of 2000 mg/kg bw.  Animals then were observed for 14 days. No mortality occurred in this limit test. There were no treatment related clinical signs, necropsy findings or changes in body weight. Slight to moderate erythema and mainly moderate edema with partly decreasing intensity were observed at the treatment sites up to day 3 and were fully reversible within 7 days. MDEA-Esterquat C16-18 and C18 unsatd. is practically non-toxic based on the dermal LD50 of > 2000 mg/kg bw determined in this study.

 

Weight of evidence

The acute oral toxicity for MDEA-Esterquat C16-18 and C18 unsatd., MDIPA-Esterquat C18 unsatd. and MDIPA-Esterquat C16-18 and C18 unsatd. is equally low for the three of them with an LD50 >> 2000 mg/kg bw. Reliable data is available for all three substances. Low dermal toxicity with an LD50 >> 2000 mg/kg bw is shown in vivo for both source substances, MDEA-Esterquat C16-18 and C18 unsatd. and MDIPA-Esterquat C18 unsatd. Together with the low dermal penetration of ≤ 1.4 % as shown for MDEA-Esterquat C16-18 and C18 unsatd. in vivo and ≤ 0.3% for MDIPA-Esterquat C16-18 and C18 unsatd. in vitro this supports the omission of testing for the target substance and permits the use of data from structurally closely related substances to fulfil the data requirements for the endpoint acute dermal toxicity.

Testing of acute dermal toxicity of MDIPA-Esterquat C16-18 and C18 unsatd. is scientifically not justified according to REACH Regulation Annex XI 1 as reliable and relevant data from two structurally related substances are used to fulfill this information requirement in a weight of evidence approach. In addition, testing via the dermal way is scientifically not necessary because reliable data on acute oral toxicity as well as oral and dermal bioavailability is available for the substance to be registered. Because the oral LD50 for female rats was > 2000 mg/kg bw and bioavailability is around 50 times higher than dermal bioavailability, acute toxicity after dermal administration is not to be expected and testing in animals is not warranted. Further support is given by retrospective data analyses undertaken by Creton et al. (2010) and Seidle et al. (2010) to ascertain the value of regulatory requirements prescribing multiroute testing for acute systemic toxicity. These analyses have examined the concordance among regulatory classifications for acute oral, dermal, and/ or inhalation toxicity for ~500 agrochemical and biocidal active substances and nearly 2000 industrial chemicals. The findings from these two independent reviews have revealed that acute dermal studies of pure substances do not add value above and beyond oral data for hazard classification of pesticides, biocides, or chemicals.

 

Acute inhalation toxicity

Conduct of an acute inhalation study is regarded unnecessary for the following reasons: MDEA and MDIPA based Esterquats share the property of having a very low vapour pressure. MDIPA-Esterquat C16-18 and C18 unsatd.has a vapour pressure of approx. 8.4E-7 Pa,excluding inhalation exposure by vapours. The generation of dusts and aerosols is prevented by appropriate RMMs, and the substance is not used in spray applications. Thus, there is no concern with respect to respiratory irritation.

 

Given that acute toxicity is unlikely to occur based on low acute toxicity via the oral and dermal route, as well as on low exposure levels, testing by the inhalation route is scientifically not necessary according to REACH Regulation Annex XI 1 and 3.

 

Based on the available information, the acute toxicity of MDIPA-Esterquat C16-18 and C18 unsatd. is low for all three routes of administration. There are no data gaps in acute toxicity. Even though there is no information on acute toxicity in humans, there is no reason to believe that the low acute toxicity observed in experimental animals would not be relevant for human health.

References

Seidle T. et al.: Cross-Sector Review of Drivers and Available 3Rs Approaches for Acute Systemic Toxicity Testing, TOXICOLOGICAL SCIENCES 116(2), 382–396 (2010).

Creton S. et al.: Acute toxicity testing of chemicals—Opportunities to avoid redundant testing and use alternative approaches, Critical Reviews in Toxicology, 2010; 40(1): 50–83

Justification for selection of acute toxicity – oral endpoint

OECD guideline study, no deviations, RL1, GLP

Justification for selection of acute toxicity – inhalation endpoint

inhalation is no relevant route of exposure

Justification for selection of acute toxicity – dermal endpoint

No single study has been selected as key study, since all available data (OECD guideline studies, no deviations, RL1, GLP) are considered together.

Justification for classification or non-classification

Based on the available data, MDIPA-Esterquat C16-18 and C18 unsatd. does not need to be classified for acute toxicity according to regulation (EC) 1272/2008 or the former European directive on classification and labelling 67/548/EEC. Thus, no labelling is required.