Reaction mass of N-(2-hydroxyethyl)alkyl] alkylamide and glycerol

Endpoint summary

Description of key information

Tthe reaction mass of N-[2-(2hydroxyethoxy)ethylacetamide and Glycerol was evaluated for repeated dose toxicity by oral route in a Combined 28-Day Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test in rats. No test-item related toxicity was observed up to the highest dose tested (1000 mg/kg).  

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 Sept 2018 - 21 Nov 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

July 2016

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA, Health Effects Test Guidelines OPPTS 870.3650, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (July 2000)

Version / remarks:

July 2000

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: other guidance as listed under Principles of method if other than guideline

Principles of method if other than guideline:

- OECD Guidelines for Testing of Chemicals, Guideline 421, Reproduction/Developmental Toxicity Screening Test (July 2016)
- The United States EPA Health Effects Test Guidelines, OPPTS 870.3550, Reproduction/Developmental Toxicity Screening Test (July 2000)
- Commission regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B.7: "Repeated Dose (28 days) Toxicity (oral)". Official Journal of the European Union No. L142 (May 2008)
- OECD Guidelines for Testing of Chemicals, Guideline 407, Repeated Dose 28-day Oral Toxicity Study in Rodents (October 2008)
- The United States EPA Health Effects Test Guidelines, OPPTS 870.3050, Repeated dose 28-day oral toxicity study in rodents (July 2000)

GLP compliance:

yes

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: Approximately 10-11 weeks (males), 13-14 weeks (females).
- Weight at study initiation: 257-305 g (males) or 203 - 231 g (females)
- Fasting period before study: no
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon cages.
Mating: Females were caged together with males on a one-to-one-basis in Macrolon cages.
Post-mating: Males were housed in their home cage with a maximum of 5 animals/cage. Females were individually housed in Macrolon cages. During the lactation phase, females were housed in Macrolon plastic cages.
During locomotor activity monitoring, animals were housed individually in a Hi-temp polycarbonate cage.
The cages contained appropriate bedding (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany).
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 46 - 76
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 19 Sept 2018 to 21 Nov 2018

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Method of formulation: Test item dosing formulations (w/w) were homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements. The dosing formulations were prepared weekly as a solution, filled out in daily portions and stored in the refrigerator. The dosing formulations were removed from the refrigerator and stirred at room temperature for at least 30 minutes before dosing. Test item dosing formulations were kept at room temperature until dosing.
Adjustment was made for specific gravity of the test item. No correction was made for the purity/composition of the test item.
Dose volume: 5 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Duplicate sets of samples (approximately 500 mg accurately weighed) for each sampling time point were sent to the analytical laboratory. For concentration analysis, results were considered acceptable if mean sample concentration results were within or equal to ± 10% of target concentration. Duplicate middle samples (approximately 500 mg accurately weighed) for each sampling time point were sent to the analytical laboratory. For homogeneity anaylsis, results were considered acceptable if the coefficient of variation (CV) of concentrations was ≤10%. Stability analyses performed previously in conjunction with the method development and validation study demonstrated that the test item is stable in the vehicle when prepared and stored under the same conditions at concentrations bracketing those used in the present study.

Duration of treatment / exposure:

Males were treated for 29 days, i.e. 14 days prior to mating, during mating and up to and including the day before scheduled necropsy. Females that delivered were treated for 50-56 days (most females) or 63 days (one female of the mid dose group and one female of high dose group), i.e. 14 days prior to mating, the variable time to conception, the duration of pregnancy and at least 13 days after delivery, up to and including the day before scheduled necropsy.
The first day of dosing was designated as Day 1. One female from the control group, one in the mid dose group and one in the high dose group were not dosed on one occasion as these females were littering at the moment of dosing. The omission of one day of dosing over a period of several weeks was not considered to affect the toxicological evaluation.

Frequency of treatment:

Once daily, 7 d/w

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Remarks:

Low dose group

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Remarks:

Mid dose group

Dose / conc.:

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

Remarks:

High dose group

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

Dose selection rationale:The dose levels were selected based on the results of a 10-day dose range finder with oral administration of Reaction mass of N-[2-(2hydroxyethoxy)ethylacetamide and Glycerol in rats, and to determine the peak effect of occurrence of clinical signs after dosing. This dose range finding study was performed with staggered start, as no in vivo data was available for the reaction mass of N-[2-(2hydroxyethoxy)ethylacetamide and Glycerol. The first dose level was 500 mg/kg bw/day (low dose group), based on an acute toxicity end point summary that supports an oral LD50 of >2000mg/kg with QSAR data and the absence of cytotoxicity in the Neural Red 3t3 cell test. The second dose level was 1000 mg/kg bw/day (high dose group), it was selected based on the lack of toxicologically relevant changes in the low dose group up to Day 6 of treatment. Mortality was observed twice daily during the study. Clinical observations were performed at least daily from days 1-10 at 0-15 minutes, 1 hour (±15 minutes) and 3 hours (± 30 minutes) after dosing. Body weights were measured on day 1 prior to dosing and on days 5 and 10. Food consumption was recorded for the high dose group over days 1-5, and for the low dose group over days 3-5 and 5-10. All animals were subjected to an external, thoracic and abdominal examination on Day 11. Animals were not deprived of food prior to necropsy. Terminal body weight, kidney and liver weight were determined at scheduled necropsy and gross lesions were recorded. Gross lesions were not retained, no organs were fixed and histopathological examination was not performed.

Positive control:

No.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS:
Yes
- Time schedule: Twice daily, in the morning and at the end of the working day.

DETAILED CLINICAL OBSERVATIONS:
Yes
- Time schedule: once daily, beginning during the first administration of the test item and lasting throughout the dosing periods up to the day prior to necropsy.

BODY WEIGHT:
Yes
- Time schedule for examinations: Males and females were weighed on the first day of treatment (prior to dosing) and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum, and during lactation on postnatal Days 1, 4, 7 and 13. Terminal body weights were recorded on the day of necropsy (fasted for males, non-fasted for females)

FOOD CONSUMPTION:
Yes. Weekly, for males and females, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17, and 20 post-coitum and during lactation on PND 1, 4, 7, and 13.

FOOD EFFICIENCY:
No.

WATER CONSUMPTION:
- Yes, on regular basis throughout the study by visual inspection of the water bottles.

OPHTHALMOSCOPIC EXAMINATION:
No

HAEMATOLOGY:
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination.
- Anaesthetic used for blood collection: Yes (iso-flurane)
- Animals fasted: Yes, males overnight (with a maximum of 24 hours). Water was provided. Females not.
- How many animals: 5 animals/sex/group
- Parameters checked were: According to test guidelines.


CLINICAL CHEMISTRY:
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination.
- Animals fasted: Yes, males overnight (with a maximum of 24 hours). Water was provided. Females no.
- How many animals: 5 animals/sex/group
- Parameters checked were: According to test guidelines.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION:
Yes
- Time schedule for examinations: During Week 4 of treatment (males) and during the last week of lactation (females, i.e. PND 8-12). After dosing, after completion of clinical observations.
- Dose groups that were examined: all (5 animals/sex/group)
- Battery of functions tested: Hearing ability, pupillary reflex, static righting reflex, and fore- and hindlimb grip strength (recorded as the mean of three measurements per animal) and locomotor activity (recording period: 1-hour under normal laboratory light conditions, using a computerized monitoring system). Total movements and ambulations were reported.

Sacrifice and pathology:

SACRIFICE
Scheduled euthanasias as follows:
- Males: after completion of mating period, minimum 28 days of administration.
- Females: PND 14-16

GROSS PATHOLOGY: Yes
- All animals were subjected to a full post mortem examination, with special attention being paid to the reproductive organs.


ORGAN WEIGHTS
- On selected 5 animals/sex/group: according to guidelines.
- All remaining animals: epididymis, prostate, seminal vesicles including coagulation glands, testes, thyroid.

HISTOPATHOLOGY: Yes
- On selected 5 animals/sex/group in control and high-dose groups and all animals that were euthanized in extremis: according to guidelines.
- On all animals: all gross lesions.
- All remaining animals, including males that failed to sire and females that failed to deliver pups: Cervix, epididymis, glands (coagulation, mammary, parathyroid, pituitary, prostate, seminal vesicle,and thyroid), gross lesions/masses, ovaries, testes, uterus, vagina.
- For the testes of all selected males of the low and high dose groups, and one male of the control group, a detailed qualitative examination was made, taking into account the tubular stages of the spermatogenic cycle.

Other examinations:

Estrous cycles were evaluated by examining the vaginal cytology of samples obtained by vaginal lavage. Daily vaginal lavage was performed for all females beginning 13 days prior to treatment, the first 14 days of treatment and during mating until evidence of copulation was observed. Vaginal lavage was continued for those females with no evidence of copulation until termination of the mating period. On the day of necropsy, a vaginal lavage was also taken to determine the stage of estrous. This was done for all females, except for one female from the control group that had to be euthanized in extremis.

Statistics:

All statistical tests were conducted at the 5% significance level. All pairwise comparisons were conducted using two sided tests and were reported at the 1% or 5% levels. Numerical data collected on scheduled occasions for the listed variables were analyzed as indicated according to sex and occasion. Descriptive statistics number, mean and standard deviation (or %CV or SE when deemed appropriate) were reported whenever possible. Inferential statistics were performed according to the comparison matrix below when possible, but excluded semi-quantitative data, and any group with less than 3 observations. The following pairwise comparisons were made: Low dose group vs. control, mid dose group vs. control, and high dose group vs. control.
Parametric: Datasets with at least 3 groups (the designated control group and 2 other groups) were compared using Dunnett-test (many-to-one-t-test).
Non-Parametric: Datasets with at least 3 groups was compared using a Steel-test (many-to-one rank test).
The motor activity data set was compared using an overall Kruskal-Wallis.
Incidence: An overall Fisher’s exact test was used to compare all groups at the 5% significance level.
The above pairwise comparisons were conducted using Fisher’s exact test whenever the overall test is significant.

Clinical signs:

no effects observed

Description (incidence and severity):

No treatment-related clinical signs were noted during daily detailed clinical observations or during weekly arena observations.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

On Day 19 of post-coitum, one female from the control group was sacrificed as she expressed severe clinical symptoms including lethargy, a flat and hunched posture, slow breathing, piloerection, hypothermia, partially closed eyes (i.e. ptosis), and a dehydrated and pale appearance. At macroscopic examination, main findings were a pale appearance of the animal, an enlarged liver, gelatinous pancreas, enlarged iliac and renal lymph nodes, and a red discolouration of the mesenteric and renal lymph nodes The uterus contained in total 12 dead fetuses. Microscopic examination revealed marked necrosis of the liver (correlating with a macroscopically enlarged liver) and marked tubular degeneration of the kidneys. These alterations in liver and kidney were regarded the main cause of moribundity for this animal.
On the day of scheduled necropsy, one female from the high dose group died after the blood sampling procedure. There were no microscopic findings of note for this animal and this death was regarded to be related to the blood sampling procedure under anaesthesia and therefore unrelated to treatment with the test item.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Mean body weight of treated females at 1000 mg/kg bw/day was statistically significantly increased (4.7%) when compared with concurrent control on Day 1 of dosing and remained slightly higher throughout treatment. As the body weight gain of these females remained similar to control values, the higher mean body weight on Day 1 was attributed to biological variation (females were allocated prior to start pretest and thus at least two weeks prior to the first administration of test item).

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption before or after correction for body weight was similar to the control level over the treatment period.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

A decrease in mean corpuscular volume (MCV) of 4% (mean 52.1 ± 2.0, n=5; control mean 54.5± 1.3, n=5 ) and in mean corpuscular haemoglobin (MCH) of 6% (mean 1.7 ± 0.04, n=5; control mean 1.14 ± 0.05, n=5) compared with the concurrent control was noted in males at 300 mg/kg bw/day. These changes were considered unrelated to administration of the test item due to the minimal magnitude of the change and absence of a dose response.
In females at 1000 mg/kg bw/day, the mean eosinophils level was statistically significantly decreased (mean 0.0 ± 0.0, n=4). As values remained within the historical control range (Historical control data for eosinophils in female Wistar Han rats period 2017-2018 (10E9/L): mean: 0.1; P5-P95: 0.00-0.13, n=206), this finding was considered unrelated to treatment.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

The statistical signifance noted for mean albumin levels of males at 100 mg/kg bw/day (mean 32.0 ± 0.7, n=5) and 1000 mg/kg bw/day (mean 32.7 ± 0.6, n=5) were attributed to a relatively high concurrent control value (control mean 33.8 ± 0.7, n=5) and therefore considered unrelated to treatment with the test item. (Historical control data for albumin (g/L) in male Wistar han rats for period 2017-2018: mean: 32.2, P5 – P95: 30.2 – 34.0, n=270).
Other statistically significant changes in clinical biochemistry parameters of males at 100 mg/kg bw/day, i.e. total protein (mean 63.0 ± 1.6, n=5; control mean 66.7 ± 2.5 n=5) and potassium (mean 3.62 ± 0.24, n=5; control mean 4.00 ± 0.20, n=5) were considered to be unrelated to treatment as these occurred in the absence of a dose-related trend.
Thyroid hormone analyses: Serum levels of T4 in F0-males were not considered to be affected by treatment.
Coagulation parameters of treated rats were considered not to have been affected by treatment.

Urinalysis findings:

not examined

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals. In males, mean grip strength of the fore legs was decreased with 11% at 1000 mg/kg bw/day (mean 1279 ± 100, n=5) when compared with concurrent control (mean 1439 ± 157, n=5). As no statistical significance was achieved, all values remained well within the historical control data range and the concurrent control mean was relatively high, this change was attributed to biological variation (Historical control data for forlimb grip strength (gram) in male Wistar Han rats period 2015-2018: mean = 1158, P5 - P95 = 681 - 1606, n=445). Grip strength values in females were unremarkable. Motor activity was similar between treated and control groups. All groups showed a similar motor activity habituation profile with a decreasing trend in activity over the duration of the test period.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

The mean relative spleen weight of females at 1000 mg/kg bw/day was deceased with 12% (mean 0.167 ± 0.007, n=4) when compared with concurrent control (mean 0.190 ± 0.028, n=5) . As no statistical significance was achieved and as all values remained well within the historical control range, this change was considered not to be toxicologically relevant. (Historical control data for relative spleen weight (%) in female Wistar han rats, period 2017-2018: mean: 0.174, P5 – P95: 0.145 – 0.210, n=205).
The statistical significance noted for the mean absolute thymus weight of females at 100 mg/kg bw/day was considered unrelated to treatment in the absence of a dose-related response.

Gross pathological findings:

no effects observed

Description (incidence and severity):

Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment. All of the recorded macroscopic findings were within the range of background gross observations encountered in rats of this age and strain.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

All of the recorded microscopic findings were within the range of background pathology encountered in rats of this age and strain. There was no test item-related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

Histopathological findings: neoplastic:

no effects observed

Key result

Dose descriptor:

NOAEL

Remarks:

Parental

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No toxicity was observed up to the highest dose level tested.

Key result

Critical effects observed:

no

Analysis of dose preparations: In the control group formulation, no test substance was detected. The concentrations analysed in the formulations of groups exposed to 100, 300 and 1000 mg/kg bw/day were in agreement with target concentrations (i.e. mean accuracies between 93% and 97%). The formulations of the low and high dose group were homogeneous (i.e. coefficient of variation ≤10%).

Conclusions:

In a repeated dose oral toxicity study combined with a reproduction/developmental toxicity screening study with rats (OECD 422), no parental toxicity was observed up to the highest dose level tested (1000 mg/kg bw day). Therefore, the no-observed-adverse-effect level (NOAEL) of the reaction mass of N-[2-(2hydroxyethoxy) ethylacetamide and Glycerol was established to be ≥ 1000 mg/kg bw/day.

Executive summary:

A repeated dose oral toxicity study combined with a reproduction/developmental toxicity screening study with rats was performed according to OECD/EC guidelines and GLP principles. Reaction mass of N-[2-2hydroxyethoxy) ethylacetamide and Glycerol was administered by daily oral gavage to male and female rats at dose levels of 100, 300 and 1000 mg/kg bw/day. The rats of the control group received the vehicle, i.e. water. Males were treated for 2 weeks prior to mating, during mating, and up to termination (for 29 days). Females that delivered offspring were treated for 2 weeks prior to mating, during mating, during post-coitum, and at least 13-15 days of lactation, for 50-56 days (most females) or 63 days (one female of the mid dose group and one female of the high dose group). Test formulations prepared were considered homogeneous at the concentrations tested and analysis of the accuracy revealed acceptable levels. No test item related mortality occurred. Treatment up to 1000 mg/kg bw/day was well tolerated as indicated by the absence of treatment related changes in any of the parental parameters examined (i.e. survival, clinical appearance, functional tests, body weight, food consumption, hematology, clinical chemistry (including serum T4 in males), macroscopic examination, organ weights, and microscopic examination). Based on the results of this study, the no-observed-adverse-effect level (NOAEL) of the reaction mass of N-[2-(2hydroxyethoxy) ethylacetamide and Glycerol was established to be at least 1000 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Experimental exposure time per week (hours/week):

7

Species:

rat

Quality of whole database:

Combined 28-day/reproduction screening study (OECD 422) with exposures of females up to 62 days (up to 19 days post-partum). Study is GLP compliant with Klimisch score 1.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Mode of Action Analysis / Human Relevance Framework

No cytotoxicity is observed at highest concentration of 10 mM after 4 hr or 24 h in in-vitro genotoxicity studies with either Human lymphocytes (OECD 487) and mouse lymphoma cells (OECD 490), and also in hCLAT assay, even 5000 µg/mL was without cytotoxicity to the human monocytic cell line THP-1. Appropriate in-vitro testing for skin and eye irritation have further shown that the substance is devoid of irritating properties.

Low reactivity, in combination with high water solubility, low Pow and low molecular weight suggest quick distribution and fast excretion after uptake, and no likelihood of accumulation.

The expected general low systemic and local toxicity that becomes apparent from profiling and in-vitro studies, is experimentally confirmed with a NOAEL for repeated dose toxicity, reproduction and developmental toxicity of 1000 mg/kg bw/day, the highest dose tested.

Additional information

ORAL

Subsequent to registration requirements not related to cosmetic application, the potential toxic effects of reaction mass of N-[2-(2hydroxyethoxy)ethylacetamide (DGA) and Glycerol was evaluated in a combined 28-day repeated dose toxicity study with reproduction/developmental toxicity screening test (OECD 422) performed under GLP. The test substance was given orally by gavage for a minimum of 28 days (males) and up to 62 days (in females, with dosing up to day 19 post partum) to Wistar Han rats, and to evaluate the potential to affect male and female reproductive performance such as gonadal function, mating behaviour, conception, parturition and early postnatal development. In addition, parental, reproduction (up to and including implantation) and developmental (from implantation onwards) effects were evaluated. The selected dose levels in this study were 0, 100, 300 and 1000 mg/kg, based on the results of the dose range finder.

The following parameters and end points were evaluated in this study: mortality/moribundity, clinical signs, functional tests, body weight and food consumption, estrous cycle length and regularity, clinical pathology, serum level of thyroid hormone T4 (F0-males), gross necropsy findings, organ weights and histopathologic examinations. In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio, live litter size and early postnatal pup development (mortality, clinical signs, body weights, anogenital distance, areola/nipple retention, serum level of thyroid hormone T4 in PND 14-16 pups and macroscopy). 

 

Formulation analyses confirmed that formulations of test item in water were prepared accurately and homogenously.

No test-item related toxicity was observed up to the highest dose tested (1000 mg/kg). Also, no reproductive toxicity or developmental toxicity was observed up to 1000 mg/kg.

 

DERMAL

No study by dermal route is available. However, considering the lower absorption by dermal route compared to oral absorption, and the overall lack of toxicity observed on oral gave study, a similar lack of toxicity can be expected from repeated dose testing via dermal application.

 

INHALATION

Exposures to humans via the inhalation route are not likely considering the low vp of 0.028 Pa at 25 °C, high boiling point of 332°C. Its use is limited to cosmetic skin care products and does not involve the forming of dusts or aerosols, particles or droplets of an inhalable size.

Considering the lack of exposure via inhalation, additional testing via inhalation route is not indicated.

Justification for classification or non-classification

Repeated dose toxicity of Reaction mass of N-[2-(2hydroxyethoxy)ethylacetamide (DGA) and Glycerol has been evaluated in a combined 28-day repeated dose toxicity study with reproduction/developmental toxicity screening test (OECD 422), and was found to be without toxicity up to the highest tested dose of 1000 mg/kg bw/day.

Consequently, the substance does not need to be classified.