2-[(2-[2-(dimethylamino)ethoxy]ethyl)methylamino]ethanol

Endpoint summary

Description of key information

Repeated dose toxicity

- oral: a key K1, 90 -day repeated dose toxicity study in male and female Wistar rats was conducted according to OECD guideline 408 (Edwards, 2012). The NOAEL was established to be greater than 100 mg/kg bw/day , the highest dose tested.

- dermal or inhalation: a key study is available for the oral route of exposure. According to the REACH Regulation, only one route of exposure should be tested for repeated dose toxicity (column 2, Annex VIII, section 8.6.1). Therefore, it is not necessary to perform a repeated dose toxicity study via the inhalation or dermal route of exposure.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-05 to 2012-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Test item: JEFFCAT ZF-10
- Chemical name: N,N,N'-Trimethyl-N'-hydroxyethyl-bis-aminoethyl ether
- Batch number: 1K703
- CAS number 83016-70-0
BIOAGRI code: AGR-1892/11
- Physical state: liquid
- Sample arrival date in BIOAGRI-DF: 28/Dec/2012
- Sample storage: in a specific room (Test Item Storage Room), at room temperature, protected from humidity and light

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
a) Species: rat (Rattus norvegicus)
b) Strain: Wistar Hannover
c) Source: BIOAGRI Laboratórios – DF/ Brazil
d) Sex: Males and females (nulliparous and non-pregnant)
e) Age (on the first day of dosing): 8 to 9 weeks old
f) Number of animals on study: 10 animals/sex/group.
- Weight : 286.0g +/- 14.2g for males and 184.4g +/- 8.2g for females after randomization
- Fasting period before study: yes
- Housing: polypropylene rodent cages (41x34x19 cm) with wire mesh tops and wood shavings as bedding material.
- Diet (e.g. ad libitum): Nuvilab CR-1 diet for rodents by Nuvital Nutrientes Ltda, Curitiba –PR, Brazil, available ad libitum
- Water (e.g. ad libitum): Filtered drinking water was supplied by CAESB (Companhia de Saneamento Ambiental do Distrito Federal) in water bottles and was available ad libitum.
- Acclimation period: minimum of 6 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.0 °C - 23.2 °C
- Humidity (%): 44.0% -70.0%
- Air changes (per hr): 10-20 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

Route of administration:

oral: gavage

Vehicle:

other: deionized water

Details on oral exposure:

Daily, by oral gavage, at approximately the same time of the day, on a 7-day-a-week-basis, for a period of 13 weeks, in a constant dosage volume of 10 mL/kg.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily

Dose / conc.:

1 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in deionized water

Dose / conc.:

10 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in deionized water

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Basis: nominal in deionized water

No. of animals per sex per dose:

10 animals/sex/group

Control animals:

yes, concurrent vehicle

Details on study design:

All animals were randomized and observations were performed in a “blinded” study design.

Observations and examinations performed and frequency:

Animals were checked twice daily for mortality or morbidity on working days and once daily on Saturdays, Sundays and public holidays.
They were subjected to a careful clinical examination before the initiation of the treatment and once a week during the dosing period.
Body weight and food consumption were recorded once a week during the study period.
An ophthalmoscopic examination was performed on all animals prior to the initiation of dosing and at the end of treatment period.
A neurotoxicological evaluation using a Functional Observation Battery was performed during study week 13.
At the end of the study, urine was collected after an overnight fasting period and urinalysis parameters were examined. Blood samples were collected for hematology, leukogram, clotting and clinical chemistry analyses.

Sacrifice and pathology:

A complete gross examination was performed on all animals after euthanasia by carbon dioxide inhalation.
At study termination, the animals were subjected to a gross examination and the appropriate organs/tissues were removed, weighed and processed for histopathological evaluation. Histopathological slides obtained from the control and high dose groups were evaluated microscopically, plus two males with macroscopic lesions (stomach and urinary bladder) at mid dose and two females at low (liver) and mid dose (ovary).

Statistics:

Quantitative variables such mean body weight and mean body weight change, food consumption, FOB, hematological assessment (hemogram, leukogram, clotting time and biochemistry) were analyzed by One-Way Analysis of Variance (ANOVA), followed by Dunnett’s test if significance is detected and mean body weight change, food consumption, FOB and hematological assessment (hemogram, leukogram, urinalysis and biochemistry), followed by Mann-Whitney U test if
significance is detected. For qualitative or non-parametric data such as urinalysis, macroscopic and microscopic findings comparison was carried out using the Chi-Square Test. The level of significance was set at 5% and the statistical program used was SAS Software (SAS Institute Inc., Cary, NC).

Clinical signs:

no effects observed

Description (incidence and severity):

The test item induced no clinical signs of toxicity.

Mortality:

no mortality observed

Description (incidence):

The test item induced no mortality.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Effects on body weight or body weight gain were not observed during this study.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Effects on food consumption were not observed during this study.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Description (incidence and severity):

There were no significant findings in the hematological, leukogram and coagulation parameters.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

There were no significant findings in the clinical chemistry parameters.

Urinalysis findings:

no effects observed

Description (incidence and severity):

There were no significant findings in the urinalysis parameters.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

There were no significant findings in the Functional Observation Battery.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Pathological changes of toxicological interest were not observed.

Gross pathological findings:

no effects observed

Description (incidence and severity):

Pathological changes of toxicological interest were not observed.

Neuropathological findings:

no effects observed

Description (incidence and severity):

There were no significant findings in the Functional Observation Battery.

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Pathological changes of toxicological interest were not observed.

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

> 100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Conclusions:

Based on the results obtained in the repeated dose 13 week oral toxicity study, the NOAEL (No Observed Adverse Effect Level) of the test substance was considered to be greater than 100 mg/kg/day in males and female Wistar rats.
This test was requested by the National Competent Authority in accordance with Article 16(1) of Directive 67/548/EEC.

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1990-08-01 to 1990-08-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): TEXACAT ZF-10
- IUPAC name: 2-[(2-[2-(dimethylamino)ethoxy]-ethyl)methylamino]ethanol
- Physical state: Clear light yellow liquid
- Analytical purity: 97%
- Expiration date of the lot/batch: 2 years
- Stability under test conditions: According to study sponsor, stable for 2 years
- Storage condition of test material: in the original container under ambient conditions in non-continuous artificial light

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Charles River Crl:CD(SD) BR VAF PLUS

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd., Margate, Kent, England
- Age at study initiation: 28 +/- 1 day
- Weight at study initiation: 68-87 g
- Housing: All rats were initially caged, as far as possible, in groups of five according to sex.
- Diet (e.g. ad libitum): Biosure LAD 1 Diet, ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.8 - 21.1
- Humidity (%): 48.6 - 55.0
- Air changes (per hr): 20
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 25 Jul 1990 To: 31 Aug 1990

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: A series of formulations were prepared weekly at concentrations of 0.1, 1.0 and 10% w/v. The solutions were stored at 4 °C and allowed to warm to room temperature immediately prior to dosing.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Test duration: 29 days

Frequency of treatment:

Dosing regimen: 7 days/week

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control

Dose / conc.:

10 mg/kg bw/day (nominal)

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5 Males and 5 females per dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: 7 day preliminary oral toxicity study carried out at HRC and acute toxicity data provided by the Sponsor

Observations and examinations performed and frequency:

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily for signs of ill health, behavioral changes or toxicosis. Twice daily for dead or moribund animals.

BODY WEIGHT: Yes
- Time schedule for examinations: Prior to dosing then at weekly intervals

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as mg food/kg body weight/day: Yes
- Time schedule for examinations: Weekly

WATER CONSUMPTION: Yes
- Time schedule for examinations: Daily

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Before termination (week 4)
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes (overnight)
- How many animals: All surviving rats
- Parameters examined: packed cell volume, haemoglobin, red blood cell count, platelet count, mean corpuscular haemoglobin concentration, mean corpuscular volume, total white blood cell count, differential white blood cell count (neutrophils, lymphocytes, eosinophils, basophils, monocytes), cell morphology, and a thrombotest was conducted.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Before termination (week 4)
- Animals fasted: Yes
- How many animals: All surviving animals
- Parameters examined: glucose, glutamic-pyruvic transaminase (alanine aminotransferase), glutamic-oxaloacetic transaminase (aspartate aminotransferase), alkaline phosphatase, total bilirubin, cholesterol, urea nitrogen, total protein, albumin, globulin, albumin/globulin ratio, sodium, potassium, calcium, chloride, inorganic phosphorus, creatinine.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
- Organs weighed: adrenals, brain, kidneys, liver, ovaries, testes (with epididymides)
- Organs examined: adrenals, aorta, brain (medullary, cerebellar and cortical sections), caecum, colon, duodenum, eyes, head, heart, ileum, jejunum, kidneys, larynx, liver, lungs, lymph nodes (cervical and mesenteric), mammary gland, oesophagus, ovaries, pancreas, pharynx, pituitary, prostate, rectum, salivary gland, sciatic nerve, seminal vesicles, skeletal muscle, skin, spleen, sternum, stomach, testes (with epididymides), thymus, thyroid (with parathyroid), tongue, trachea, urinary bladder, uterus, vagina, and any other macroscopically abnormal tissue

HISTOPATHOLOGY: Yes
- Organs examined: adrenals, heart, kidneys, liver, lungs (some animals), ovaries (some animals), spleen, and any other macroscopically abnormal tissue
- Microscopic examinations were carried out for: all animals that died during the study; all rats of Group 1 (control group) and Group 3 (intermediate dosage group) and all surviving animals of Group 4 (high dosage group) killed on Day 30; lungs from all animals in the control, low and intermediate dosage groups and two surviving rats in the high dosage group; and ovaries from all animals in the control and intermediate dosage groups.

Statistics:

Appropriate statistical analyses were reported.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

All animals dosed at 100 and 1000 mg/kg/day showed increased salivation occasionally during week 1. These animals showed piloerection from day 11 or in one animal from day 8, accompanied or subsequently followed by hunched posture, abnormal gait (waddling), increased respiration, lethargy and pallor of the extremities. Red-brown staining of the periorbital region consistent with chromodacryorrhea and similar staining in the perinasal region was also observed. The two surviving male rats in the high dose group showed piloerection from day 11, accompanied in one of these animals by hunched posture (intermittent), waddling and in the last week by pallor of the extremities and increased respiration.
In the 100 mg/kg/day group, increased salivation following dosing was noted occasionally during Week 1 for male and female rats. For two male rats, small sores and scabs were noted on the shoulder region from Day 16 to termination; this finding however was probably incidental.
No clinical signs were noted for male and female rats treated at 10 mg/kg/day or from the control group.

Mortality:

mortality observed, treatment-related

Description (incidence):

In the 1000 mg/kg dose group, three out of 5 male rats and all five female rats died or were killed in extremis between days 10 and 21 of the study. No deaths occurred in lower dose groups.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight gain was reduced by 21% in females in the 100 mg/kg dose group. No toxicologically significant findings were noted at 10 mg/kg/day.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Food consumption was reduced in females in the 100 mg/kg dose group.

Food efficiency:

not examined

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Among animals given 1000 mg/kg/day, increased white cell counts and lymphocyte counts (by about 38% compared to controls at 4 weeks) were noted on day 16. At this time an increased platelet count (by about 35% compared to controls at 4 weeks) was noted in one of the surviving males and females. At the end of the study no significant haematological findings were noted among surviving animals
of this group. A low packed cell volume (PCV), haemaglobin and high mean corpuscular volume (MCV) were recorded at Day 16 for the three surviving male rats from the high dosage group. High platelet counts were also recorded for one male and one female for this group.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Clinical chemistry findings revealed increases in glutamic pyruvic transaminase (by about 50% compared to controls at 4 weeks) and increases in glutamic oxaloacetic transaminase by about 50% to 200%) in surviving rats of the high dose group on day 16 and at the end of the study. For all rats treated at 1000 mg/kg/day sampled on Day 16, high cholesterol and low total protein levels were recorded. No significant changes in clinical chemistry values were noted in animals of the intermediate and low dose group.Among animals given 100 mg/kg/day a decrease in the white cell count and lymphocyte count (by about 25%) compared to controls was noted in the females.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Among animals of the high dose group which died or were killed during the study congested lungs, enlarged cervical lymph nodes and an enlarged pale or pitted liver were generally found at necropsy.
An increase in the liver and kidney weight (by about 75% and 25% respectively) was seen in the two surviving males of the high dose/group. Enlarged pale livers were found in both of these animals, with one animal also showing enlarged pale kidneys.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Among animals of the high dose group which died or were killed during the study congested lungs, enlarged cervical lymph nodes and an enlarged pale or pitted liver were generally found at necropsy.
An increase in the liver and kidney weight (by about 75% and 25% respectively) was seen in the two surviving males of the high dose/group. Enlarged pale livers were found in both of these animals, with one animal also showing enlarged pale kidneys.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Microscopically there was marked centrilobular and moderate periportal hepatocyte vacuolation with perivascular oedema and inflammatory cell infiltration noted in the two surviving males of the high dose group. Minimal centrilobular hepatocyte vacuolation was noted in one male of the 100 mg/kg/day dosage group. In the kidneys, areas of marked vacuolation and enlargement of the cortical tubular epithelium were seen in the two surviving males of the high dose group. Minimal fine vacuolation of the inner cortical tubular epithelium was seen in one of the surviving males of the high dose group, and in two males given 100 mg/kg/day.
Similar changes were seen in 1 of 3 decedent males and 3 of 5 decedent females of the high dose group.

In the lymph nodes, foamy macrophages were noted in the paracortex regions in the two surviving males of the high dose group. Among decedents of both sexes from the high dose group foamy macrophages were noted in the paracortex of the spleen and/or lymph nodes.


In the lungs, foamy alveolar macrophages were noted in one surviving male of the high dose group and vacuolation of the bronchiolar epithelium in both surviving males of this group. These changes were note together with perivascular inflammatory cell infiltration and oedema in all decedent animals of this 1000 mg/kg/day dosage group.

In the stomach, focal necrosis of the lining mucosa was noted in one surviving male of the high dose group, together with vacuolation of the mucous neck cells in both surviving males of this group. Similar changes affecting the gastric mucosa were noted in 2 of 3 decedent males and all decedent females of the high dose group.

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

body weight and weight gain

food consumption and compound intake

haematology

histopathology: non-neoplastic

Key result

Dose descriptor:

NOEL

Effect level:

10 mg/kg bw/day (nominal)

Key result

Critical effects observed:

no

Conclusions:

The no effect level of the test substance was found to be 10 mg/kg/day. Within the context of this study, treatment-related effects observed at 100 mg/kg/day were not considered to represent serious damage to health. The NOAEL is established to be 100 mg/kg bw/day. The test substance is not classified as STOT RE according to the criteria laid down in the CLP Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Both experimental tests were run under GLP conditions and according to EU and/or OECD Guidelines with no major deviations identified.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

other justification

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: dermal

Data waiving:

other justification

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity - oral:

One 28 day and one 90 day repeated-dose toxicity studies for test material are available.

In the 28day repeated dose toxicity study performed according to OECD Guideline 407, 10 rats (5 females, 5 males) were exposed to 10, 100 and 1000 mg/kg/day (Huntsman 1991). For rats receiving the test substance at 1000 mg/kg/day, many treatment-related changes were seen. These included effects on clinical condition, bodyweight, food consumption, clinical pathology, macroscopic and microscopic pathology and mortalities occurred. Treatment-related changes recorded for rats treated at 100 mg/kg/day included lower bodyweight gains and food consumption for both male and female rats, decreased white blood cell and lymphocyte count for female rats, minimal centrilobular hepatocyte vacuolation in one male rat and fine vacuolation of the cortical tubular epithelium of the kidneys of two male rats. Within the context of this study, treatment-related effects observed at 100 mg/kg/day were not considered to represent serious damage to health and the NOAEL was determined to be 100 mg/kg bw/day.

A 90-day repeated dose toxicity study was performed for test substance according to OECD Guideline 408 (Huntsman, 2013). 10 rats (5 females, 5 males) were exposed to 1, 10 and 100 mg/kg/day, by gavage. Test substance induced no test item-related mortality or clinical signs of toxicity. Effects on food consumption, body weight and body weight gain were not observed during this study. There were no significant findings in the Functional Observation Battery evaluation, and in the hematological, leukogram, coagulation, clinical chemistry and urinalysis parameters. Pathological changes of toxicological interest were not observed. Under the repeated dose 13- week oral toxicity study on rats, the NOAEL (No Observed Adverse Effect Level) of test substance was considered to be greater than 100 mg/kg/day in males and female Wistar rats.

Repeated dose toxicity - inhalation:

A key study is available for the oral route of exposure. According to the REACH Regulation, only one route of exposure should be tested for repeated dose toxicity (column 2, annex VIII, section 8.6.1). Therefore, it is not necessary to perform a repeated dose toxicity study via the inhalation route of exposure.

Repeated dose toxicity - dermal:

A key study is available for the oral route of exposure. According to the REACH Regulation, only one route of exposure should be tested for repeated dose toxicity (column 2, annex VIII, section 8.6.1). Therefore, it is not necessary to perform a repeated dose toxicity study via the dermal route of exposure.

Justification for classification or non-classification

Based on the results of the oral repeated dose studies, the appropriate NOAEL value for use in the assessment is greater than 100 mg/kg bw/day. This value does not meet the criteria for classification under the CLP Regulation (EC) No 1272/2008, and therefore no classification is warranted for repeated dose toxicity (STOT RE).