Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

- Subacute (28 day) study; oral (gavage), rat (Crl:WI(Han); 3/sex/dose), similar to OECD guideline 407, GLP, bridging study; NOAEL(fertility) = 1000 mg/kg bw/d; read-across from MDIPA Esterquat C18 unsatd.

- Subacute (28 day) study; oral (gavage), rat (Charles River CD; 25/sex/dose), OECD guideline 407, GLP; NOAEL = 500 mg/kg bw/d (highest dose level tested); read across from MDEA-Esterquat C16-18 and C18 unsatd.

- Subchronic (91 day) study; oral (gavage), rat (Crl CD BR; 15/sex/dose), OECD guideline 408, GLP; NOAEL = 500 mg/kg bw/d (highest dose level tested); read across from MDEA-Esterquat C16-18 and C18 unsatd.

- Subchronic (91 day) study; oral (drinking water), rat (Fischer 344/DuCrl; 10/sex/dose), equivalent to OECD guideline 408, GLP; NOAEL(reproduction) = 1000 mg/kg bw/d (as DIPA), corresponding to 5700 mg/kg bw/d (as MDIPA-Esterquat C16-18 and C18 unsatd.); read-across from Diisopropanolamine (DIPA)

No adverse test substance-related effects were found for any fertility-related parameter measured, especially regarding organ weights of ovary and testes and histopathology of gonads, uterus, mammary gland, prostate and seminal vesicle.

Link to relevant study records

Referenceopen allclose all

Endpoint:
toxicity to reproduction
Remarks:
other: Repeated Dose 28-Day Oral Toxicity in Rodents
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1992-07-29 to 1992-08-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents) (27 July 1995)
Deviations:
yes
Remarks:
highest recommended dose of 1000 mg/kg bw was not included in this study, not all necessary organ weights were taken e.g. heart, thymus, spleen were not evaluated
GLP compliance:
yes
Species:
rat
Strain:
other: Charles River CD
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
water
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
Daily, 7 days/week for 4 weeks
Remarks:
Doses / Concentrations:
1, 10, 500 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: No significant treatment-related effects on reproductive organs (organ weights of ovary and testis and histopathology of gonads) were seen at the highest dose administered.
Reproductive effects observed:
not specified

In this Repeated Dose, 28-day Oral Toxicity Study performed comparable to OECD guideline 407, 12 May 1981 reaction products of C16-18/C18 unsaturated fatty acid with methyl diethanolamine, MeCl quaternized (10 % a.i aqueous dispersion) was administered to groups of 25 Charles River CD rats male and female by oral gavage at dose levels of 1, 10 and 500 mg/kg bw/day for 28 days. Two other control groups received either deionised water or to pH 2.5 adjusted (HCL) deionised water (vehicle).

All animals survived to study termination. No test substance-related findings were detected or observed in clinical examinations, body weights, food consumption values, opthalmoscopic examinations, haematology, clinical biochemistry or clinical pathology evaluations including organ weights of ovary and testis and histopathology of reproductive organs (gonads, mammary gland (femals only), prostate and seminal vesicle, uterus and vagina).

The no effect level (NOEL) for this study is the high dose level of 500 mg/kg bw/day of the test article.

Conclusions:
The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads from this 28-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.
Executive summary:

In this Repeated Dose, 28-day Oral Toxicity Study performed comparable to OECD guideline 407, 12 May 1981 rMDEA-Esterquat C16-18 and C18 unsatd.  (10 % a.i aqueous dispersion) was administered to groups of 25 Charles River CD rats male and female by oral gavage at dose levels of 1, 10 and 500 mg/kg bw/day for 28 days. Two other control groups received either deionised water or to pH 2.5 adjusted (HCL) deionised water (vehicle).

All animals survived to study termination. No test substance-related findings were detected or observed in clinical examinations, body weights, food consumption values, opthalmoscopic examinations, haematology, clinical biochemistry or clinical pathology evaluations including organ weights of ovary and testis and histopathology of reproductive organs (gonads, mammary gland (femals only), prostate and seminal vesicle, uterus and vagina).

The no effect level (NOEL) for this study is the high dose level of 500 mg/kg bw/day of the test article.

Endpoint:
toxicity to reproduction
Remarks:
other: Repeated Dose 90-Day Oral Toxicity in Rodents
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1992-10-30 to 1993-02-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl CD BR
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
water
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily, 7 days/week, for 13 weeks
Remarks:
Doses / Concentrations:
1, 10, 500 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
15
Control animals:
yes, concurrent vehicle
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: There were no significant adverse test substance-related effects on reproductive organs (organ weights of ovary and testis and histopathology of gonads).
Reproductive effects observed:
not specified
Conclusions:
The results from the evaluation of reproductive organs, especially organ weights of ovary and testis and histopathology of gonads from this 91-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 500 mg/kg bw/day.
Executive summary:

In a subchronic toxicity study comparable to OECD guideline 408, MDEA-Esterquat C16-18 and C18 unsatd.  (10 % a.i.) was administered to 15 Charles River CD rats / sex/ dose by gavage at dose levels of 1, 10 and 500 mg/kg bw/ day for a period of 13- weeks. One control group received the vehicle, deionized water, and a second control group received pH-adjusted, deionized water (pH 2.5). The regimen for both control groups was identical to treatment groups.

The following parameters were monitored during the study: clinical observations (detailed, weekly; mortality, morbidity, and overt signs of toxicity, twice a day); body weights (weekly); food consumption (weekly); clinical pathology (haematology, blood chemistry, and urinalysis; at termination); opothalmoscopic examinations (once pre-test and prior to sacrifice); macroscopic pathologic examination; absolute and relative organ weights; microscopic pathology.

The following reproductive organs were included in the examinations: absolute and relative organ weigths of ovary and testis and histopathology of gonads, mammary gland (females only), prostate and seminal vesicle, uterus with cervix and vagina.

There were no changes in any of these parameters that were considered to be toxicologically significant or test-substance related.

The no effect level (NOEL) for this study is the high dose level of 500 mg/kg bw/day of the test article.

Endpoint:
toxicity to reproduction
Remarks:
other:
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
2012-09-28 to 2012-11-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Similar to guideline study, GLP
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
other: EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral)) (2008)
Deviations:
yes
Remarks:
only 3 animals per sex per dose (bridging study)
Qualifier:
equivalent or similar to guideline
Guideline:
other: OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents) (2008)
Deviations:
yes
Remarks:
only 3 animals per sex per dose (bridging study)
Principles of method if other than guideline:
As this study was mainly intended for justification of read-across to a structurally comparable substance, the guidelines are not fully applicable. Only 3 animals/sex/dose have been tested. Additional male reproductive parameters have been evaluated.
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
other: Crl:WI(Han)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Groups 1 and 2: Approximately 6 weeks. Groups 3 and 4: Approximately 8 weeks.
- Weight at study initiation: not given
- Fasting period before study: no
- Housing: Group housing of 3 animals per sex in Macrolon cages (MIV type, height 18 cm) with sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lilico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: At least 5 days before the start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 to 70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 28 September 2012 To: 05 November 2012
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on exposure:
VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed and on information from the sponsor.
- Amount of vehicle (if gavage): 5 mL/kg
Details on mating procedure:
no mating; only examination of re
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily, 7 days per week
Remarks:
Doses / Concentrations:
0, 150, 500, 1000 mg/kg bw/day
Basis:
actual ingested
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: end of treatment
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 3 / sex / dose group
- Parameters checked in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: end of treatment
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 3 / sex / dose group
- Parameters checked in table 1 were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
- Organ Weights
- Staging of spermatogenesis
Oestrous cyclicity (parental animals):
no
Sperm parameters (parental animals):
Parameters examined in male parental generations:
testis weight, staging of spermatogenesis
Litter observations:
n/a
Postmortem examinations (parental animals):
GROSS PATHOLOGY: Yes (see table 2)
HISTOPATHOLOGY: Yes (see table 2)
Postmortem examinations (offspring):
n/a
Reproductive indices:
n/a
Offspring viability indices:
n/a
Dose descriptor:
NOEL
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: clinical chemistry
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
act. ingr.
Sex:
male/female
Reproductive effects observed:
not specified
Conclusions:
The results from the evaluation of reproductive organs, especially organ weights of testis, histopathology of gonads, including spermatogenic staging profiles from this 28-day repeated dose study revealed no indications of any substance-related effects up to and including the highest test dose of 1000 mg/kg bw/day.
The NOEL in this study based on clinical chemistry was determined to be 500 mg/kg bw/d, the NOAEL was determined to be 1000 mg/kg bw/d.
Executive summary:

In a subacute toxicity study similar to OECD guideline 407 (2008) and EU method B.7 (2008)MDIPA Esterquat C18 unsatd. (100%)was administered to 3 Crl:WI(Han) rats/sex/dosein propylene glycol by gavageat dose levels of 0, 150, 500, 1000 mg/kg bw/day for 28 consecutive days. As this study was mainly intended for justification of read-across to a structurally comparable substance, the guidelines are not fully applicable. Additional male reproductive parameters have been evaluated.

No toxicologically relevant changes were noted in any of the following parameters investigated in this study: clinical appearance, body weight, food consumption, haematology parameters, macroscopic examination, organ weights, and microscopic examination. The spermatogenic staging profiles were normal in the high dose group males.

Changes considered to be treatment-related were confined to higher alanine aminotransferase activity in two males and one female at 1000 mg/kg bw/d and in one male at 500 mg/kg bw/d, and higher aspartate aminotransferase activity in one male each at 500 and 1000 mg/kg bw/d.

The toxicological relevance of these increases was unclear, since based on the parameters assessed in this study it could not be determined whether these changes were indicative of liver damage or reflected adaptive hepatic enzyme induction.

The NOEL in this study based on clinical chemistry was determined to be 500 mg/kg bw/d. No effects on the reproductive organs were noted up to and including 1000 mg/kg bw/d.

Endpoint:
toxicity to reproduction
Remarks:
other: Repeated Dose 90-Day Oral Toxicity in Rodents
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study, GLP
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Fischer 344/DuCrl
Sex:
male/female
Route of administration:
oral: drinking water
Vehicle:
water
Details on mating procedure:
animals were not mated
Duration of treatment / exposure:
13 weeks;
recovery groups (control + high dose group): 13 weeks treatment + 4 additional weeks without treatment
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 100, 500, or 1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
No changes in organ weights of ovaries, testes, epididymis, uterus and no histopathological findings in gonads and secondary sex organs were reported.
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Remarks:
DIPA
Sex:
male/female
Basis for effect level:
other: minor changes in clinical chemistry in males of the 1000 mg/kg bw/d dose group; no changes in the assessed reproductive parameters
Reproductive effects observed:
not specified
Conclusions:
In this 13 week drinking water study in Fischer 344/DuCrl rats, no changes in organ weights of ovaries, testes, epididymis, uterus and no histopathological findings in gonads and secondary sex organs were reported.
The NOAEL for reproduction in this study was therefore 1000 mg/kg bw/d This corresponds to a NOAEL of 5700 mg/kg bw/d in terms of MDIPA-Esterquat C16-18 and C18 unsatd.
Executive summary:

In a subchronic toxicity study similar to OECD guideline 408 Diisopropanolamine (DIPA) (98.8 to 99.6%) was administered to 10 Fischer 344/DuCrl rats/sex/dose in drinking water at dose levels of 0, 100, 500, or 1000 mg/kg bw/d daily for 13 weeks. Additional 10 animals in the control and highest dose groups were were maintained on untreated drinking water for 4 weeks as recovery group.

No mortality occurred in any group; no treatment related clinical signs were observed. No relevant effects on body weight, ophthalmoscopic examinations, haematological parameters, prothrombin times, gross or histopathology were detected. The water consumption in the high dose group males and females was slightly reduced compared to control animals. Consistent with this, rats of the 1000 mg/kg bw/day dose group had increased urine specific gravity and females had decreased urine volume, which were also considered secondary adaptive effects.

There were minimal, but statistically identified, differences noted for cholesterol (increased), albumin (decreased) and phosphorus (decreased) for males given 1000 mg/kg bw/day. There were no effects upon clinical chemistry parameters for males given 100 or 500 mg/kg bw/day or for females at any dose level.

Increased relative kidney weights for males and females of the 500 or 1000 mg/kg bw/day dose group were observed without any treatment-related gross or histopathologic correlate.

In the recovery group, water consumption in the treated group was below control by app. the same degree as at the end of the dosing period; all other affected endpoints showed reversibility. The absolute and relative kidney weights of the treated rats were still above the controls, but the differences were about one-half of those immediately following 13 weeks dosing. No treatment-related renal histopathological changes were observed.

No changes in organ weights of ovaries, testes, epididymis, uterus and no histopathological findings in gonads and secondary sex organs were reported.

The NOAEL for general toxicity in this study was 500 mg/kg bw/d based on minor changes in clinical chemistry in males of the 1000 mg/kg bw/d dose group. The NOAEL for reproduction was 1000 mg/kg bw/d.

Based on the molecular weights of DIPA (133.19 g/mol) and MDIPA-Esterquat C16-18 and C18 unsatd. (approx. 760 g/mol), the NOAEL of 1000 mg/kg bw/d in terms of DIPA has been extrapolated to 5700 mg/kg bw/d in terms of MDIPA-Esterquat C16-18 and C18 unsatd.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
OECD guideline studies, no deviations, RL1-2, GLP
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Data on reproductive toxicity is not available for the target substance MDIPA-Esterquat C16-18 and C18 unsatd.

For the assessment of effects of MDIPA-Esterquat C16-18 and C18 unsatd. to fertility, a 28 day bridging study with the source substance MDIPA Esterquat C18 unsatd. as well as a subacute 28 day oral toxicity study and a subchronic 91 day oral toxicity study conducted with the source substance MDEA-Esterquat C16-18 and C18 unsatd. (Reaction products of C16-18/C18 unsaturated fatty acid with methyl diethanolamine, MeCl quaternised), are available.

 

In the bridging study MDIPA Esterquat C18 unsatd. was administered to 3 Crl:WI(Han) rats/sex/dose in propylene glycol by gavage at dose levels of 0, 150, 500, 1000 mg/kg bw/day for 28 consecutive days (for detailed study descriptions see section “Repeated dose toxicity”). As this study was mainly intended for justification of read-across to a structurally comparable substance, the guidelines are not fully applicable. Additional male reproductive parameters have been evaluated. No toxicologically relevant changes were noted in any of the following parameters investigated in this study: clinical appearance, body weight, food consumption, haematology parameters, macroscopic examination, organ weights, and microscopic examination. The spermatogenic staging profiles were normal in the high dose group males. No effects on the reproductive organs were noted up to and including 1000 mg/kg bw/d.

 

In the repeated dose toxicity studies with the source substance MDEA-Esterquat C16-18 and C18 unsatd. , Charles River CD rats (Sprague-Dawley) were treated with the test substance by oral gavage for 28 days (25 animals/sex/dose) or for 91 days (15 animals/sex/dose) at dose levels of 0, 1, 10, or 500 mg/kg bw/day (for detailed study descriptions see section “Repeated dose toxicity”). In both studies, no adverse test substance-related effects were found for any parameter measured, especially regarding organ weights of ovary and testes and histopathology of gonads, uterus, mammary gland, prostate and seminal vesicle.

Further testing for effects concerning fertility is considered not to be necessary because up to 500 mg/kg bw/day there were no indications for substance-related effects in general and especially regarding organ weights (ovary and testis) and histopathology of gonads from the repeated dose toxicity studies. In addition this is substantiated by the absence of effects on maternal reproduction, embryo lethality or embryotoxicity in the developmental toxicity study in rats with doses of up to 1000 mg/kg bw/day.

 

In a subchronic toxicity study similar to OECD guideline 408 Diisopropanolamine (DIPA) (98.8 to 99.6%) was administered to 10 Fischer 344/DuCrl rats/sex/dose in drinking water at dose levels of 0, 100, 500, or 1000 mg/kg bw/d daily for 13 weeks. Additional 10 animals in the control and highest dose groups were were maintained on untreated drinking water for 4 weeks as recovery group.

No mortality occurred in any group; no treatment related clinical signs were observed. No relevant effects on body weight, ophthalmoscopic examinations, haematological parameters, prothrombin times, gross or histopathology were detected. The water consumption in the high dose group males and females was slightly reduced compared to control animals. Consistent with this, rats of the 1000 mg/kg bw/day dose group had increased urine specific gravity and females had decreased urine volume, which were also considered secondary adaptive effects.

There were minimal, but statistically identified, differences noted for cholesterol (increased), albumin (decreased) and phosphorus (decreased) for males given 1000 mg/kg bw/day. There were no effects upon clinical chemistry parameters for males given 100 or 500 mg/kg bw/day or for females at any dose level.

Increased relative kidney weights for males and females of the 500 or 1000 mg/kg bw/day dose group were observed without any treatment-related gross or histopathologic correlate.

In the recovery group, water consumption in the treated group was below control by app. the same degree as at the end of the dosing period; all other affected endpoints showed reversibility. The absolute and relative kidney weights of the treated rats were still above the controls, but the differences were about one-half of those immediately following 13 weeks dosing. No treatment-related renal histopathological changes were observed.

No changes in organ weights of ovaries, testes, epididymis and uterus and no histopathological findings in gonads and secondary sex organs were reported.

The NOAEL for general toxicity in this study was 500 mg/kg bw/d based on minor changes in clinical chemistry in males of the 1000 mg/kg bw/d dose group. The NOAEL for reproduction was 1000 mg/kg bw/d.

Based on the molecular weights of DIPA (133.19 g/mol) and MDIPA-Esterquat C16-18 and C18 unsatd. (approx. 760 g/mol), the NOAEL of 1000 mg/kg bw/d in terms of DIPA has been extrapolated to 5700 mg/kg bw/d in terms of MDIPA-Esterquat C16-18 and C18 unsatd.

 

The performance of a two-generation reproductive toxicity study is not necessary in accordance with Annex XI (3.2 (a)) of REACH Regulation (EC) No 1907/2006 based on a quantitative assessment. The results of exposure and risk assessments covering all relevant exposures throughout the life cycle of the substance demonstrate a low exposure and a RCR value below 1 in all scenarios of manufacture, formulation, professional use, consumer use and indirect exposure of humans via the environment (see Chapter 9 and 10 of the CSR). 

Strictly conservative DNELs have been derived from results of the available test data which include a pre-natal developmental study and sub acute and sub-chronic studies with evaluation of reproductive endpoints. No substance-related adverse effects were found in any of the tests conducted and the NOAELs used to derive the DNELs correspond to the maximum doses tested.  

The DNELs fertility have been derived from results of the sub-chronic repeated dose toxicity study, taking full account of the potential increased uncertainty resulting from the omission of the information requirement by applying an additional assessment factor. These derived DNELs are relevant and appropriate both for the information requirement to be omitted and for risk assessment purposes.

The exposure and risk assessments are included in chapters 9 and 10 of this CSR. The final conclusion is based on the risk characterisation ratio (RCR). Comparison of all the derived DNELs with the results of the exposure assessment shows that exposures in all life cycle stages of the substance are well below the derived DNELs even under the precautionary assumptions applied.

 

There are no data gaps for effects on fertility. No human data are available. However, there is no reason to believe that these results from rat would not be applicable to humans.

Endpoint specific justification for read-across

For details on substance identity, toxicokinetics and detailed toxicological profiles, please refer also to the general justification for read-across given in chapter 5 of the CSR and attached as pdf document to section 7 of the IUCLID file.

Structural similarity

a. Structural similarity and functional groups

The target substance, MDIPA-Esterquat C16-18 and C18 unsatd., consists of an amine backbone (MDIPA = Methyldiisopropanol amine) esterified with long chain fatty acids C16, C18 and C18 unsaturated. The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Methosulfate.

The first source substance, MDEA-Esterquat C16-18 and C18 unsatd. , consists of an amine backbone (MDEA = Methyldiethanol amine) esterified with long chain fatty acids C16, C18 and C18 unsaturated. The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The second source substance, MDIPA-Esterquat C18 unsatd., consists of an amine backbone (MDIPA = Methyldiisopropanol amine) esterified with long chain fatty acids C18 unsaturated. The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Methosulfate.

The source and the target substances share structural similarities with common functional groups (quaternary amines), esters, and fatty acid chains with comparable length and degree of saturation. The amine backbones based on MDEA and MDIPA, respectively, differ only by one methyl group, all functional groups are identical.

 

b. Common breakdown products

The metabolism expected to occur is hydrolysis of the ester-bond by esterases. However, the rate of hydrolysis is assumed to be low. The fraction of metabolised molecules would result in free fatty acids and Dimethyl-DEA (DEA = Diethanolamine) and Dimethyl-DIPA (DIPA = Diisopropanolamine), respectively. The carboxylic acids are further degraded by the mitochondrial beta-oxidation process (for details see common text books on biochemistry). The fatty acids enter normal metabolic pathways and are therefore indistinguishable from fatty acids from other sources including diet. The quaternary ammonium ions are not expected to be further metabolised, but excreted unchanged via the urine.

 

c. Differences

The differences in fatty acid chain length (higher percentage of C16 in the source substance MDEA-Esterquat C16-18 and C18 unsatd. ) and degree of saturation (higher degree of unsaturation in the source substance MDIPA-Esterquat C18 unsatd.) may be relevant for local effects (e.g. irritation) but are not considered to be of relevance for reproductive toxicity.

Chloride is an essential nutrient and present in all organisms; excess chloride is renally excreted (see common textbooks on biology / biochemistry). Methyl sulphate is metabolised to Sulphate and Carbon    dioxide, and these are excreted via the urine and released by the lungs, respectively. The anions Chloride and Methyl sulphate are not expected to have any influence on toxicity or reactivity.

The methyl side chain of Dimethyl-DIPA which is not present in Dimethyl-DEA is not expected to enhance reactivity, which is supported by a similar toxicological profile for the source and target substance as well as toxicokinetic data for DEA (Diethanolamine), MDEA, DIPA (Diisopropanolamine) and MDIPA (for details see general justification for read-across).

 

Description of interpolation based on structures and supporting amine data (see attachment)

 

 

Source

substance 1

Target substance

Source

substance 2

Supporting data

MDEA-Esterquat C16-18 and C18 unsatd.

MDIPA-Esterquat C16-18 and C18 unsatd.

 

MDIPA-Esterquat C18 unsatd.

 

Diisopropanolamine

(DIPA)

 

 

OECD guideline 407, rat, oral; NO(A)EL 500 mg/kg bw/day (highest tested dose)


OECD guideline 408, rat, oral; NO(A)EL 500 mg/kg bw/day (highest tested dose)

 

OECD guideline 407, rat, oral
bridging study with 3 animals/sex/dose
NOEL 500 mg/kg bw/day; NOAEL 1000 mg/kg bw/day

OECD guideline 408, rat, drinking water;

 

NOAEL(reproduction) = 1000 mg/kg bw/d as DIPA, corresponding to 5700 mg/kg bw/d asMDIPA-Esterquat C16-18 and C18 unsatd.

 

 

 

 

Fatty Acid Chain length distribution

<C16 <7%

C16, 16‘, 17 26-35%

C18 42-52%

C18‘ 15-20%

C18‘‘, 18‘‘‘ </= 1.5%

>C18 </= 2%

< C16: <= 6%

C16 - < C18: 20-65%

C18: 4-60%

C18 unsatd.: 10-43%

> C18: <= 5%

<C16 -

C16 </=7%

C18 </= 4%

C18‘ 55-65%

C18‘‘ 18-25%

C18‘‘‘ 6-12%

  >C18 </= 5%

Not applicable

Amine

MDEA

MDIPA

MDIPA

DIPA

Anion

Chloride

Methyl sulphate

Methyl sulphate

Not applicable

 

 

 

 

 

 

 

The interpolation of the available data on repeated dose toxicity to fulfil the information requirement for the target substance is based on:

-      identical fatty acid chain in the source substance MDEA-Esterquat C16-18 and C18 unsatd. and the target substance MDIPA-Esterquat C16-18 and C18 unsatd.

-      identical amine headgroup and counter ion in the source substance MDIPA-Esterquat C18 unsatd. and the target substance MDIPA-Esterquat C16-18 and C18 unsatd.

 

In the repeated dose toxicity studies with the source substance MDEA-Esterquat C16-18 and C18 unsatd. no adverse test substance-related effects were found for any parameter measured, especially regarding organ weights of ovary and testes and histopathology of gonads, uterus, mammary gland, prostate and seminal vesicle. The 28-d bridging study with MDIPA Esterquat C18 unsatd. also included histopathological examinations of the reproductive organs and staging of spermatogenesis. The spermatogenic staging profiles were normal in the high dose group males. No effects on the reproductive organs were noted up to and including 1000 mg/kg bw/d.

The read-across approach is further substantiated by the data on the repeated dose toxicity of DIPA. No data is available for the potential metabolite Dimethyl-DIPA itself. To support the absence of toxicity, data on DIPA , which is a structural part of the target substance MDIPA-Esterquat C16-18 and C18 unsatd. and the source substance MDIPA Esterquat C18 unsatd., is provided. No treatment related effects on reproductive parameters were reported up to the limit dose of 1000 mg/kg bw/d in terms of DIPA.

 

Quality of the experimental data of the analogues:

Fertility:

The source substance MDEA-Esterquat C16-18 and C18 unsatd. has been tested in reliable GLP-compliant OECD TG 407 and OECD TG 408 studies up to 500 mg/kg bw/d.Fertility-related parameters were assessed: organ weights of ovary and testes and histopathology of gonads, uterus, mammary gland, prostate and seminal vesicle.

 

The source MDIPA Esterquat C18 unsatd. has been tested in a GLP compliant 28 d bridging study similar to OECD TG 407 up to 1000 mg/kg bw/d. As this study was mainly intended for justification of read-across to a structurally comparable substance, only 3 animals/sex/dose have been tested. Additional male reproductive parameters have been evaluated.

 

DIPA has been tested in a GLP compliant study (well-documented publication) equivalent to OECD guideline 408 up to the limit dose of 1000 mg/kg bw/d.

 

The available data from the source substances are sufficiently reliable to justify the read-across approach.

 

Implications of differences in classification and labelling for read-across

The source substance MDEA-Esterquat C16-18 and C18 unsatd. is not classified for any human health hazard. The second source substance MDIPA Esterquat C18 unsatd. is classified for local effects (irreversible effects on the eye Category 1, irritating to the skin Category 2) and the target substance MDIPA-Esterquat C16-18 and C18 unsatd. is also classified for local effects (irritating to the eye Category 2, irritating to the skin Category 2). These differences are not considered relevant forreproductive toxicity.

 

Conclusion for read-across

The structural similarities between the source and the target substances and the similarities in their breakdown products presented above support the read-across hypothesis. Adequate and reliable scientific information indicates that the source and target substances and their subsequent degradation products have similar toxicity profiles.

The dose descriptor obtained from the existing sub-chronic repeated dose toxicity study performed on the source substance MDEA-Esterquat C16-18 and C18 unsatd. is considered as an appropriate starting point for deriving a DNEL for the target substanceMDIPA Esterquat C16-18 and C18 unsatd.The remaining uncertainty associated with this read-across approach is accounted for by using the appropriate assessment factors, as recommended in ECHA Guidance R.8.

Effects on developmental toxicity

Description of key information

Prenatal developmental toxicity study; oral (gavage); rat (Wistar, 25/group, dosed from day 6 through 15 post coitum); OECD Guideline 414; GLP; NOEL = 1000 mg/kg bw/d, read across from MDEA-Esterquat C16-18 and C18 unsatd.

Prenatal developmental toxicity study; oral (gavage); rat (CRL:CD(SD), 25/group, dosed from day 6 through 20 post coitum); equivalent to OECD Guideline 414; GLP; NOAEL = 1000 mg/kg bw/d, supporting information from Diisopropanolamine (DIPA)

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1992-10-09 to 1992-11-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
other: Rat WIST HanIbm: WIST (SPF)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd.
- Age at study initiation: No data. (Age at pairing: 12 weeks, minimum).
- Weight at study initiation: No data. (189-228 g at day 0 post coitum).
- Fasting period before study: N/A.
- Housing: individually in Makrolon cages (type-3) with wire mesh tops and standarized granulated softwood bedding (Lignocel, Schill AG, CH 4132 Muttenz/Switzerland).
- Diet (e.g. ad libitum): pelleted standard Kilba 343 rat/mouse maintenance diet ("Kilba", Klingentalmuehle AG, CH 4303 Kaiseraugust/Switzerland) ad libitum. (Batch no. 65-92).
- Water (e.g. ad libitum): tap water in bottles ad libitum.
- Acclimation period: 10 days (minimum).


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 (air-conditioned).
- Humidity (%): 40-70.
- Air changes (per hr): 10-15.
- Photoperiod (hrs dark / hrs light): 12-hrs artificial fluorescent light/12 hours dark.


IN-LIFE DATES: From: 1992-10-09 To: 1992-11-21
Route of administration:
oral: gavage
Vehicle:
other: NOAEL emBe-distelled water, previously adjusted with hydrochloric acid to pH 2.5-2.6
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The mixtures of the test substance and vehicle were prepared daily before administration. The test substance was weighed into a glass beaker on a tared precision balance (Mettler PE 360) and the vehicle added (w/v). The mixtures were prepared using a homogenizer. During the daily administration period, homogeneity was maintained using a magnetic stirrer. All animals received a dose volume of 10 ml/kg bw with a daily adjustment of the individual volume to the actual body weight.


DIET PREPARATION
- Rate of preparation of diet (frequency): N/A.
- Mixing appropriate amounts with (Type of food): N/A.
- Storage temperature of food: N/A.


VEHICLE: Bi-distilled water (previously adjusted with hydrochloric acid to pH 2.5-2.6).
- Justification for use and choice of vehicle (if other than water): N/A.
- Concentration in vehicle: N/A.
- Amount of vehicle (if gavage): 10 ml/kg.
- Lot/batch no. (if required): N/A.
- Purity: N/A.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of the test substance/vehicle dilutions were taken on two occasions during the dosing period of the study and sent to the Sponsor for analysis (concentration, homogeneity and stability over 2-hrs).
Details on mating procedure:
- Impregnation procedure: cohabitation.
- If cohoused:
- M/F ratio per cage: 1:1.
- Length of cohabitation: overnight
- Further matings after two unsuccessful attempts: no data.
- Verification of same strain and source of both sexes: no data. (The male rats used for mating were in the possession of the testing laboratory. The fertility of these males was proved and was continuously controlled).
- Proof of pregnancy: sperm in vaginal smear/vaginal plug referred to as day 0 post coitum.
- Any other deviations from standard protocol: After mating, female rats were removed and allocated to the test groups.
Duration of treatment / exposure:
Day 6 through 15 post coitum.
Frequency of treatment:
Once daily in the morning.
Duration of test:
Females were sacrificed on day 21 post coitum.
Remarks:
Doses / Concentrations:
0 mg/kg bw/day (vehicle control)
Basis:
actual ingested
Remarks:
Doses / Concentrations:
50 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
250 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
25/group.
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dosages were based on the results of the dose range-finding sudy (RCC Project 326158--see cross reference section).
- Rationale for animal assignment (if not random): Mated rats were assigned to the different groups using a computer-generated random algorithm.
- Rationale for method of administration: International guidelines recognize the efficacy of oral administration.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes.
- Time schedule: The animals were checked at least twice daily for any mortalities. The animals were observed at least twice daily for signs of reaction to treatment and/or symptoms of ill health.


DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recordered daily, from day 0 until day 21 post coitum.
- Body weight gains from days 0-6 p.c., 6-9 p.c., 9-12 p.c., 12-16 p.c., 6-16 p.c., 16-21 p.c., and 6-21 p.c. were calculated.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes. Food consumption was recorded for the following periods: days 0-6, 6-9, 9-12, 12-16, and 16-21 post coitum.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food consumed per period/days per period: Yes.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


POST-MORTEM EXAMINATIONS: Yes. Any female sacrificed or found dead during the study was subjected to gross macroscopic examination of all internal organs.
- Sacrifice on day # 21 post coitum by CO2 asphyxiation. Fetuses were removed by Caesarean section.
- Organs examined: Emphasis on the uterus and its contents, position of the fetuses in the uterus, and number of corpora lutea.
- Specimens of abnormal tissue were fixed in neutral phosphate buffered 4% formaldehyde solution.


Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes.
- Gravid uterus weight: Yes. (The uteri (and contents) of all females with live fetuses were weighed at necropsy on day 21 post coitum to enable the calculation of the corrected body weight gain).
- Number of corpora lutea: Yes.
- Number of implantations: Yes. If no implantation sites were evident, the uterus was placed in an aqueous solution of ammonium sulfide to accentuate possible haemorrhagic areas of implantation sites.
- Number of early resorptions: Yes.
- Number of late resorptions: Yes.
- Other: position of the fetuses was recordered.
Fetal examinations:
The fetuses were removed from the uterus, sexed, weighed individually, examnined for gross external abnormalities and allocated to one of the following procedures: Wilson's slicing technique or skeletal examination.

- External examinations: Yes: all per litter.
- Soft tissue examinations: Yes: half per litter. Wilson's slicing technique for examination of the viscera and brain. One half of the live fetuses from each litter were fixed in a mixture of ethyl alcohol, formol and acetic acid. After examination, the sections were preserved in a solution of ethyl alcohol and glycerine (one fetus/container).
- Skeletal examinations: Yes: half per litter. Fetuses were placed in a solution of potassium hydroxide for clearing and stained with alizarin red S (modified technique). The skeletons were examined and all abnormalities were recordered. The specimens were preserved individually in plastic bags.
- Head examinations: No data.
Statistics:
The following statistical methods were used to analyze body weights, food consumption, reproduction and skeletal examination data: means and standard deviations, univariate one-way analysis of variance, Dunnett-test, Steel-test, Fisher's Exact test for 2x2 tables. Individual values, means, standard deviations and t-statistics were rounded off before printing.
Indices:
N/A.
Historical control data:
N/A.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
MORTALITY: No test substance related deaths were noted. One female in group 2 (50 mg/kg), was found dead in the evening of day 10 post coitum. At first daily inspection in the morning, this female had ruffled fur, ataxia, tremor and bleeding from the vagina. This single finding in the low dose
group was considered to be incidental.

CLINICAL SIGNS OF REACTION TO TREATMENT: In group 4 (1000 mg/kg), one female had bleeding from the vagina on days 15-16 post coitum. At scheduled Caesarean section, this female had one empty implantation site, only. No abnormal findings were noted in the dams of groups 1 (0 mg/kg), 2 (50 mg/kg), or 3 (250 mg/kg) with the exception of the dam in group 2 (50 mg/kg) mentioned above.

FOOD CONSUMPTION: Up to and including the highest dose level of 1000 mg/kg, food consumption of the dams was similar in all groups. No test substance related effects were noted.

BODY WEIGHTS: The differences in mean body weights between the control group and any dose group did not reveal any test substance related effect. In all groups the body weight gain and the corrected body weight gain (corrected for uterus weight) were similar. The dams of group 4 (1000 mg/kg), had statistically significantly increased mean body weights on day 5, between days 7-13 and on day 15 post coitum. These findings were considered to be a consequence of the slightly increased initial mean body weight: 213 g compared to 207 g in the control group, on day 0 post coitum.

REPRODUCTION DATA: Values for post-implantation loss were: 4.8 and 8.7 in groups 1 (0 mg/kg) and 4 (1000 mg/kg), respectively. The latter value attained statistical significance. Although the post-implantation losses were within the normal range of the historical control data of this strain of animals, two additional females (which were not included in the statistical analysis) had total post-implantation losses (i.e. implantation sites, only). Therefore, the increased post-implantation loss in group 4 (1000 mg/kg) was considered to be a slight effect of treatment with the test substance. In groups 2 (50 mg/kg) and 3 (250 mg/kg), none of the reproduction parameters as assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and mean number of fetuses per dam were adversely affected by treatment with the test substance.

NECROPSY FINDINGS: No test substance related abnormal findings were noted at terminal necropsy of the dams in any group. One female in group 2 (50 mg/kg) which was found dead on day 10 post coitum had reddish lungs with dark red foci, reduced right kidney and enlarged left kidney with nodules and calcus in pelvis (both kidneys with pelvic dilation) and in the urinary badder dark red discoloration of the mucosa and urine with hemorrhagic contents were noted. No abnormal findings were noted in the dams of groups 1 (0 mg/kg), 3 (250 mg/kg), or 4 (1000 mg/kg).
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
EXTERNAL EXAMINATION: No test substance related abnormal findings were noted. Neither the type nor the incidences of the abnormal findings listed below indicated test substance related effects: In each group 1 (0 mg/kg) and 3 (250 mg/kg), one fetus with reduced mouth and shortened and tapering lower jaw was noted. In group 2 (50 mg/kg), one runt (fetal weight < 2.5 g) was noted. In group 4 (1000 mg/kg), one fetus with several abnormal findings--generally oedematous, cranioschisis with excencephaly, protusion of the tongue, eventration and both eyes partially missing--was noted.

SEX RATIOS: In all groups, the sex ratios of the fetuses were not affected by the treatment with the test substance.

BODY WEIGHTS: The mean fetal body weights were similar in all groups. No test substance related effects were evident.

VISCERAL EXAMINATION BY WILSON TECHNIQUE: No test substance related abnormal findings were noted at visceral examination of the fetuses. In group 3 (250 mg/kg), in the fetus which had reduced mouth, shortened and tapering lower jaw at external examination a small palatoschisis was additionally observed at visceral examination. No abnormal findings were noted in groups 1 (0 mg/kg), 2 (50 mg/kg), or 4 (1000 mg/kg).

SKELETAL EXAMINATION: The incidences of abnormal findings at skeletal examination were as following:
2/145 (=1%) fetuses in 2/25 litters in group 1 (0 mg/kg);
10/141 (=7%) fetuses in 6/24 litters in group 2 (50 mg/kg);
4/136 (=3%) fetuses in 4/25 litters in group 3 (250 mg/kg) and
4/116 (=3%) fetuses in 4/21 litters in group 4 (1000 mg/kg).
Mainly: wavy or fused ribs, abnormally or incompletely ossified sternebrae, dumbbell shaped or hemicentric thoracic vertebral body and in two fetuses in group 2 (50 mg/kg) fused os interparietale and occipitale were noted. Neither the type nor the incidences of these abnormal findings indicated test substance related effects. In all groups, the stage of the skeletal development of the fetuses was similar. The few statistically significant differences (on individual basis, only) between the control group and the dose group 4 (1000 mg/kg) in the ossification grade of the phalangeal nuclei did not indicate any test substance related effect. All values evaluated in this study were in the normal range of the historical control data of this strain of animals.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: fetotoxicity
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Basis for effect level:
other: teratogenicity
Abnormalities:
no effects observed
Developmental effects observed:
no

N/A.

Conclusions:
NOAEL = 1000 mg/kg bw/day (general tolerability in the females and for the fetal organism).
NOAEL = 1000 mg/kg bw/day (maternal reproduction).
NOAEL = 1000 mg/kg/day (teratologic effects)
Executive summary:

In the developmental toxicity study (OECD guideline 414), groups of 25 mated female Wistar rats were treated with the test substance MDEA-Esterquat C16-18 and C18 unsatd. orally by gavage once daily from day 6 through 15 post coitum, at dose levels of 0, 50, 250 and 1000 mg/kg bw/day. Females were sacrificed on day 21 post coitum and the fetuses were removed by Caesarean section.

At 50, 250, and 1000 mg/kg, for the dams no test substance-related deaths or clinical signs as were noted as reaction to treatment. Up to and including the highest dose level of 1000 mg/kg, food consumption and body weight development of the dams were not affected by treatment with the test substance. At necropsy, no test substance-related abnormal findings in the dams were noted in any group.

A slight (statistically non significant) decrease in the number of corpora lutea and a slight (statistically non significant) increase in pre-implantation losses were observed in the high dose group (17.4 %) as compared to the controls (12.8 %); however, this is not a substance-related effect, since exposure started at gestation day 6, the day of implantation.

The slightly but statistically significant increased post-implantation losses at the high dose of (8.7 %) as compared to the controls (4.8 %) resulted in a slightly reduced portion of total fetuses per implantation site (91.3%) and in a reduced mean litter size (10.5 fetuses/litter) as compared to the controls (95.2 %; 11.2 fetuses/litter). The values were within the range of historical control values (3.9 % to 11.6 % for post-implantation losses and 10.2 to 12.2 fetuses/litter).

At 1000 mg/kg, two females with total post-implantation losses were noted. One of the two females had bleeding from the vagina on days 15-16 post coitum. However these two animals had only two and one corpora lutea, respectively, and were obviously not fit for reproduction. A small but significant increase in post-implantation losses was noted for the remaining females; however, the increase was within the historical data of the laboratory. These findings were considered by the authors to be a potentially effect of the test substance.

As the values were well within the range of the historical control values recorded at the same laboratory, it is likely that the effects observed are incidental and therefore not treatment related.

At 50 and 250 mg/kg, no test substance-related effects were noted on the maternal reproductive parameters, assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and by the mean number of fetuses per dam.

Up to and including the highest dose level of 1000 mg/kg, no adverse effects on the fetal parameters were recorded. No external, skeletal or soft tissue malformations and no external variations were found. Mean fetal body weights and the sex ratios of the fetuses were comparable in all groups.

The maternal NOAEL is 1000 mg/kg bw/day. 

The developmental NOAEL is 1000 mg/kg bw/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
OECD guideline study, no deviations, RL1, GLP
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Prenatal developmental toxicity data on the target substance MDIPA-Esterquat C16-18 and C18 unsatd. is not available.

For the assessment of prenatal developmental toxicity of the target substance MDIPA-Esterquat C16-18 and C18 unsatd. a prenatal developmental toxicity study with the source substance MDEA-Esterquat C16-18 and C18 unsatd. and supporting information from the amine DIPA are available.

In a study according to OECD guideline 414, groups of 25 mated female Wistar rats were treated with MDEA-Esterquat C16-18 and C18 unsatd. orally by gavage once daily from day 6 through 15 post coitum, at dose levels of 0, 50, 250 and 1000 mg/kg bw/day. Females were sacrificed on day 21 post coitum and the fetuses were removed by Caesarean section.

At 50, 250, and 1000 mg/kg bw/d, for the dams no test substance-related deaths or clinical signs as were noted as reaction to treatment. Up to and including the highest dose level of 1000 mg/kg bw/d, food consumption and body weight development of the dams were not affected by treatment with the test substance. At necropsy, no test substance-related abnormal findings in the dams were noted in any group.

A slight (statistically non significant) decrease in the number of corpora lutea and a slight (statistically non significant) increase in pre-implantation losses were observed in the high dose group (17.4 %) as compared to the controls (12.8 %); however, this is not a substance-related effect, since exposure started at gestation day 6, the day of implantation.

The slightly but statistically significant increased post-implantation losses at the high dose of (8.7 %) as compared to the controls (4.8 %) resulted in a slightly reduced portion of total fetuses per implantation site (91.3%) and in a reduced mean litter size (10.5 fetuses/litter) as compared to the controls (95.2 %; 11.2 fetuses/litter). The values were within the range of historical control values (3.9 % to 11.6 % for post-implantation losses and 10.2 to 12.2 fetuses/litter).

At 1000 mg/kg bw/d, two females with total post-implantation losses were noted. One of the two females had bleeding from the vagina on days 15-16 post coitum. However these two animals had only two and one corpora lutea, respectively, and were obviously not fit for reproduction. A small but significant increase in post-implantation losses was noted for the remaining females; however, the increase was within the historical data of the laboratory. These findings were considered by the authors to be a potentially effect of the test substance. At 50 and 250 mg/kg bw/d, no test substance-related effects were noted on the maternal reproductive parameters, assessed by the mean number per dam of corpora lutea and implantation sites, pre- or post-implantation losses, and by the mean number of fetuses per dam.

Up to and including the highest dose level of 1000 mg/kg bw/d, no adverse effects on the fetal parameters were recorded. No external, skeletal or soft tissue malformations and no external variations were found. Mean fetal body weights and the sex ratios of the fetuses were comparable in all groups.

There are indications that the slightly increased incidences of post-implantation losses at 1000 mg/kg bw/d are due to some maternal (toxic) effects, which could not be further evaluated because this dose level has not been tested in the repeated dose toxicity studies.

The slightly increased post-implantation losses at the high dose compared to the controls could be caused by some maternal toxicity, incidentally and therefore not treatment related or through a direct toxic effect on the fetus. Since two females of the high dose had total post-implantation losses (implantation-sites only) and one of these females showed vaginal bleeding, this may indicate that the post-implantation losses are due to (some) maternal toxicity effects. Other long-term studies with this high dose level are not available to further evaluate substance-related toxicity. Therefore, the impact of the maternal effects cannot be evaluated in depth. As the values were well within the range of the historical control values recorded at the same laboratory, it is likely that the effects observed are incidental and therefore not treatment related. Since there was no effect on fetal body weight and no increased incidences of abnormal fetuses, it is assumed that the post-implantation losses are not due to a direct effect on the fetus.

The results of the prenatal developmental toxicity study do not indicate a substance-related effect on the fetus up to the limit dose of 1000 mg/kg bw/d. Therefore, the aspect of prenatal developmental toxicity is sufficiently covered.

 

In a developmental toxicity study similar to OECD Guideline 414 Diisopropoanolamine (98.8 to 99.6%) was administered to 25 female time-mated CRL:CD(SD) rats/dose by gavage at dose levels of 0, 100, 300, or 1000 mg/kg bw/day from day 6 through 20 of gestation.

No effects were observed with respect to clinical signs, body weights, body weight gains, food consumption, organ weights or gross pathology at necropsy.

There were no treatment-related effects on pregnancy rates, resorption rates, litter size, number of corpora lutea or implantations, percent pre-implantation loss, percent post-implantation loss, fetal body weights or gravid uterine weights.

There were no statistically identified differences in the incidence of any fetal alteration in any of the treated groups compared to controls. The small number of alterations observed in fetuses from dams given DIPA either occurred at low frequencies and/or not in a dose-related manner.

The maternal and developmental NOAEL was 1000 mg/kg bw/d.

Based on the molecular weights of DIPA (133.19 g/mol) and MDIPA-Esterquat C16-18 and C18 unsatd. (approx. 760 g/mol), the NOAEL of 1000 mg/kg bw/d in terms of DIPA has been extrapolated to 5700 mg/kg bw/d in terms of MDIPA-Esterquat C16-18 and C18 unsatd.

 

Performance of a prenatal developmental toxicity study in a second species is not necessary in accordance with Annex XI (3.2 (a)) of REACH Regulation (EC) No 1907/2006 based on a quantitative assessment. The results of exposure and risk assessments covering all relevant exposures throughout the life cycle of the substance demonstrate a low exposure and a RCR value below 1 in all scenarios of manufacture, formulation, professional use, consumer use and indirect exposure of humans via the environment. The very small spectrum of identified uses as referred to in Annex VI section 3.5 is documented in chapters 9 and 10 in this CSR. The exposure assessment and risk characterisation according to Article 14(4) and Annex I, 5 of REACH Regulation (EC) No 1907/2006 have been conducted solely for the purpose of justifying this waiver, given that the substance is not classified as dangerous or as PBT/vPvB according to any of the Regulation’s criteria.

Strictly conservative DNELs have been derived from results of the available test data which include a pre-natal developmental study and sub acute and sub-chronic studies with evaluation of reproductive endpoints. No substance-related adverse effects were found in any of the tests conducted and the NOAELs used to derive the DNELs correspond to the maximum doses tested. The DNELs fertility have been derived from results of the sub-chronic repeated dose toxicity study, taking full account of the potential increased uncertainty resulting from the omission of the information requirement by applying an additional assessment factor.DNELs for developmental toxicity derived from the developmental toxicity study were higher than the DNELs for fertility. Thus, the fertility DNELs are also protective for development.These derived DNELs are relevant and appropriate both for the information requirement to be omitted and for risk assessment purposes.

The exposure and risk assessments are included in chapters 9 and 10 of this CSR. The final conclusion is based on the risk characterisation ratio (RCR). Comparison of all the derived DNELs with the results of the exposure assessment shows that exposures in all life cycle stages of the substance are well below the derived DNELs even under the precautionary assumptions applied.

 

There are no data gaps for effects on development. No human data are available. However, there is no reason to believe that these results from rat would not be applicable to humans.

Endpoint specific justification for read-across

For details on substance identity, toxicokinetics and detailed toxicological profiles, please refer also to the general justification for read-across given in chapter 5 of the CSR and attached as pdf document to section 7 of the IUCLID file.

Structural similarity

a. Structural similarity and functional groups

The target substance, MDIPA-Esterquat C16-18 and C18 unsatd., consists of an amine backbone (MDIPA = Methyldiisopropanol amine) esterified with long chain fatty acids C16, C18 and C18 unsaturated. The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Methosulfate.

The source substance, MDEA-Esterquat C16-18 and C18 unsatd. , consists of an amine backbone (MDEA = Methyldiethanol amine) esterified with long chain fatty acids C16, C18 and C18 unsaturated. The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source and the target substances share structural similarities with common functional groups (quaternary amines), esters, and fatty acid chains with comparable length and degree of saturation. The amine backbones based on MDEA and MDIPA, respectively, differ only by one methyl group, all functional groups are identical.

 

b. Common breakdown products

The metabolism expected to occur is hydrolysis of the ester-bond by esterases. However, the rate of hydrolysis is assumed to be low. The fraction of metabolised molecules would result in free fatty acids and Dimethyl-DEA (DEA = Diethanolamine) and Dimethyl-DIPA (DIPA = Diisopropanolamine), respectively. The carboxylic acids are further degraded by the mitochondrial beta-oxidation process (for details see common text books on biochemistry). The fatty acids enter normal metabolic pathways and are therefore indistinguishable from fatty acids from other sources including diet. The quaternary ammonium ions are not expected to be further metabolised, but excreted unchanged via the urine.

 

c. Differences

Chloride is an essential nutrient and present in all organisms; excess chloride is renally excreted (see common textbooks on biology / biochemistry). Methyl sulphate is metabolised to Sulphate and Carbon dioxide, and these are excreted via the urine and released by the lungs, respectively. The anions Chloride and Methyl sulphate are not expected to have any influence on toxicity or reactivity.

The methyl side chain of Dimethyl-DIPA which is not present in Dimethyl-DEA is not expected to enhance reactivity, which is supported by a similar toxicological profile for the source and target substance as well as toxicokinetic data for DEA (Diethanolamine), MDEA, DIPA (Diisopropanolamine) and MDIPA (for details see general justification for read-across).

 

Description of read-across based on structures and supporting amine data (see attachment)

 

 

Source substance

Target substance

Supporting data

MDEA-Esterquat C16-18 and C18 unsatd.

MDIPA-Esterquat C16-18 and C18 unsatd.

 

Diisopropanolamine

(DIPA)

 

 

OECD Guideline 414, rat, oral (gavage), day 6-15 p.c.; NOEL = 1000 mg/kg bw/d (highest tested dose)

 

OECD Guideline 414, rat, oral (gavage), day 6-20 p.c.; NOAEL = 1000 mg/kg bw/d as DIPA (highest tested dose),corresponding to 5700 mg/kg bw/d asMDIPA-Esterquat C16-18 and C18 unsatd.

 

 

Fatty Acid Chain length distribution

<C16 <7%

C16, 16‘, 17 26-35%

C18 42-52%

C18‘ 15-20%

C18‘‘, 18‘‘‘ </= 1.5%

>C18 </= 2%

< C16: <= 6%

C16 - < C18: 20-65%

C18: 4-60%

C18 unsatd.: 10-43%

> C18: <= 5%

Not applicable

Amine

MDEA

MDIPA

DIPA

Anion

Chloride

Methyl sulphate

Not applicable

 

The read-across of the available data on prenatal developmental toxicity to fulfil the information requirement for the target substance is based on:

-      identical fatty acid chain in the source substance MDEA-Esterquat C16-18 and C18 unsatd. and the target substance MDIPA-Esterquat C16-18 and C18 unsatd.

The read-across approach is further substantiated by data on prenatal developmental toxicity of DIPA. No data is available for the potential metabolite Dimethyl-DIPA itself. To support the absence of toxicity, data on DIPA, which is a structural part of the target substance MDIPA-Esterquat C16-18 and C18 unsatd., is provided.

In both prenatal developmental toxicity studies, withthe source substance MDEA-Esterquat C16-18 and C18 unsatd. as well as with the structural element DIPA, no adverse test substance related effects were noted up to the limit dose of 1000 mg/kg bw/d.

Moreover, as described in more detail in the general justification for read-across, source and target substance were neither genotoxic, nor sensitisers. Both outcomes further support the lack of reactivity towards cellular macromolecules.

 

Quality of the experimental data of the analogues

The source substance MDEA-Esterquat C16-18 and C18 unsatd. has been tested in reliable GLP-compliant OECD TG 414 study up to the limit dose of 1000 mg/kg bw/d. There are no uncertainties, thus the study can be used in an analogue approach.

DIPA has been tested in a GLP compliant study (well-documented publication) equivalent to OECD TG 414 up to the limit dose of 1000 mg/kg bw/d.

The available data from the source chemical are sufficiently reliable to justify the read-across approach.

 

Implications of differences in classification and labelling for read-across

The source substance MDEA-Esterquat C16-18 and C18 unsatd. is not classified for any human health hazard, whereas the target substance MDIPA-Esterquat C16-18 and C18 unsatd. is classified for local effects (irritating to the eye Category 2, irritating to the skin Category 2). This is not relevant for the endpoint that is read across (prenatal developmental toxicity).

 

Conclusion

The structural similarities between the source and the target substances and supporting data on the amines presented above support the read-across hypothesis. Adequate and reliable scientific information indicates that the source and target substances and their subsequent degradation products have similar toxicity profiles: low systemic toxicity, not mutagenic, no effects on fertility.

The absence of prenatal developmental toxicity for the source substance MDEA-Esterquat C16-18 and C18 unsatd. is also relevant for the target substance MDIPA-Esterquat C16-18 and C18 unsatd. based on the close similarities in the toxicological profiles.

As genotoxicity is one possible mechanism for developmental toxicity, the negative outcome of the genotoxicity tests for both, source and target substance, further supports the read-across. Genotoxicity is based on covalent binding of the substance itself or reactive metabolites to cellular macromolecules as rate determining step. A similar mechanism based on reactive metabolites is involved in sensitisation; both substances were not sensitisers. Thus, the consistency across these endpoints increases the confidence in the conclusion that there is no concern for reactive metabolites. 

Overall it can be concluded that the read-across approach is justified and no classification of MDIPA-Esterquat C16-18 and C18 unsatd. for developmental toxicity is required.

Justification for classification or non-classification

Results of the prenatal developmental toxicity study conducted with the source substance MDEA-Esterquat C16-18 and C18 unsatd. do not indicate a substance-related effect on the fetus up to the limit dose of 1000 mg/kg bw/day. The slightly increased incidences of post-implantation losses at 1000 mg/kg bw/day are within the range of historical control data and might therefore be accidental and not treatment related. Vaginal bleeding observed in one animal with total post-implantation losses might be due to some maternal (toxic) effects which cannot be further evaluated. Therefore, the aspect of prenatal developmental toxicity is sufficiently covered and the substance does not require to be labelled for developmental toxicity.

Results of the repeated dose toxicity tests conducted with the source substance MDIPA Esterquat C18 unsatd. as well as with the source substance MDEA-Esterquat C16-18 and C18 unsatd. do not indicate a substance-related effect on reproduction (fertility) up to the dose of 500 mg/kg bw/day, which was the highest dose tested in the full subacute and subchronic toxicity tests.

In conclusion, results of existing studies indicate that MDIPA-Esterquat C16-18 and C18 unsatd. does not need to be classified for effects on toxicity to reproduction according to Directive 67/548/EEC as well as on toxicity to reproduction (fertility and development) according to GHS Regulation EC No 1272/2008 and therefore labelling is not necessary.

Additional information