2,2',2''-nitrilotriethanol

Endpoint summary

Description of key information

oral: In a sub-chronic oral toxicity study, a NOAEL of 1000 mg/kg bw/day was established, the highest dose tested.

dermal: In a sub-chronic dermal toxicity study, NOAELs of 125 and 250 mg/kg bw/day were established for local effects for males and females. Systemic NOAELs of 125 and 250 mg/kg bw/day were determined for males and females, respectively, based on kidney effects. Similar effects were observed in a sub-chronic dermal study in mice, performed according to the same protocol.

inhalation: In a sub-acute inhalation toxicity study with rats, a NOAEC for systemic effects of 0.5 mg/L was established, the highest dose tested. 0.02 mg/L (the lowest dose tested) was considered to be the NOAEC for local effects in females. Since slight local effects were observed in males, this concentration was determined to be the LOAEC for local effects in males.

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:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study which meets basic scientific principles

Qualifier:

equivalent or similar to guideline

Guideline:

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

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: Cox CD

Sex:

male/female

Details on test animals or test system and environmental conditions:

20 male and 20 female per group
Diet: ad libitum

Route of administration:

oral: feed

Vehicle:

other: standard chow

Duration of treatment / exposure:

91 days

Frequency of treatment:

continuously

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

Dose / conc.:

500 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

20

Control animals:

yes, concurrent no treatment

Details on study design:

Post-exposure period: no

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
FOOD EFFICIENCY: Yes
HISTOPATHOLOGY: Yes
HAEMATOLOGY: Yes

Sacrifice and pathology:

HISTOPATHOLOGY: Yes

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

One rat escaped from its cage during the third week of the test and had to be eliminated because of contamination. One male from the mid-dose group and one female from the low-dose group displayed symptoms of inner ear infection.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The only abnormality observed was a slightly elevated WBC in one animal. This change was not considered significant. All other values are within normal limits and there were no significant differences between treatment groups.

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

There were no significant differences notes in organ to body weight ratios.

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

No lesions were found which followed a pattern of agent-induced toxicity. Tissue alterations observed were mild and were not considered significant.

Details on results:

AVERAGE BODY WEIGHT GAIN, FEED CONSUMPTION, AND EFFICIENCY
The only adverse significant differences noted were in body weight gain and feed efficiency in the female rats of the mid-dose group.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: no significant effects at highest dose tested

Key result

Critical effects observed:

not specified

Bodyweight gain

	males	females
Control	341±30	157±23
TEA-1 (250 mg/kg)	339±35	170±13
TEA-1 (500 mg/kg)	331±28	151±15
TEA-1 (1000 mg/kg)	329±32	161±22

 

Feed consumption

	males	females
Control	2143±155	1542±105
TEA-1 (250 mg/kg)	2073±165	1592±108
TEA-1 (500 mg/kg)	2076±282	5166±115
TEA-1 (1000 mg/kg)	2091±155	1604±123

 

Feed efficiency

	males	females
Control	15.95±0.98	10.18±1.1
TEA-1 (250 mg/kg)	16.35±0.97	10.72±0.67
TEA-1 (500 mg/kg)	15.56±0.84	8.62±2.7
TEA-1 (1000 mg/kg)	15.75±0.99	9.99±0.88

   

Organ to bodyweight ratio (LIVER)

	males	females
Control	38.2±3.3	42.1±12.2
TEA-1 (250 mg/kg)	41.6±3.7	39.9±3.3
TEA-1 (500 mg/kg)	45.9±6.9	44.6±1.0
TEA-1 (1000 mg/kg)	49.6±2.9	45.3±2.9

 

Organ to bodyweight ratio (KIDNEY)

	males	females
Control	7.6±0.6	8.1±0.3
TEA-1 (250 mg/kg)	9.3±1.3	10.2±1.4
TEA-1 (500 mg/kg)	9.2±0.7	11.1±0.7
TEA-1 (1000 mg/kg)	10.4±1.4	10.3±1.4

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

similar to OECD TG 408

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 17 - September 05 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Principles of method if other than guideline:

Groups of 10 Wistar rats/sex were head-nose exposed to clean air (control ) and target concentrations of 0.02; 0.1 and 0.5 mg/L Triethanolamine (test groups). The substance was administered as a liquid aerosol during a period of 28 days (20 exposures). Based on the recommendation of OECD Guidelines for the "Testing of Chemicals", method 412, clinical, hematological, clinicochemical examinations were carried out. Furthermore, neurofunctjonal tests (functional observational battery) were performed before and 4 times at about weekly intervals during exposure period. At the end of the study 3 animals/sex and group were perfusion fixed for eventual further neuropathology which, however, was not performed because no neurotoxic findings were observed clinically and in routine histopathology of brain and sciatic nerve. The remaining 7 animals/sex and group were subjected to conventional necropsy. 7 animals per sex of the high concentration group and of the controls were subjected to hi stopathol ogi cal examination, the scope of organs of OECD 412 being extended by brain and peripheral nerve.

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Dr. K. Thomae GmbH, Biberach- Age at study initiation: 7 weeks- mean weight at study initiation: males: 239; females 168 g- Fasting period before study: none- Housing: individually- Diet: KLIBA rat/mouse/hamster Iaboratory diet 24-343-4 10 mm pellets, KlingentalmuehIe AG, Kaiseraugst, Switzerland ad libitum- Water: tap water ad libitumENVIRONMENTAL CONDITIONS- Temperature (°C): 20-24- Humidity (%): 30-70- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation

Type of inhalation exposure:

nose/head only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

Generation of an inhalation atmosphere: The substance to be tested was supplied to a two-component atomizer by means of a continuous infusion pump and was atomized with compressed air. Having passed a glass separator, the liquid aerosol was diluted with conditioned blast air which was conducted via a glass bottle filled with bidist. water for humidification and was supplied to the exposure apparatus. In order to decrease the viscosity of the substance the two-component atomizers were warmed. The following amounts of air were set:
Test group Compressed air [m³/h] Blast air [m³/h] Exhaused air [m³/h]0-3.0 ± 0.32.7 ± 0.311.5 ± 0.3 1.5 ± 0.32.7 ± 0.321.5 ± 0.3 1.5 ± 0.32.7 ± 0.331.5 ± 0.3 1.5 ± 0.32.7 ± 0.3
Test group Substance flow atomizer [mL/h] Atomizer temperature [° C]10.2 38.9 – 41.221.2 – 4.0 38.7 – 39.9320 - 35 38.2 – 39.9
Generation of an inhalation atmosphere: The substance to be tested was supplied to a two-component atomizer by means of a continuous infusion pump and was atomized with compressed air. Having passed a glass separator, the liquid aerosol was diluted with conditioned blast air which was conducted via a glass bottle filled with bidist. water for humidification and was supplied to the exposure apparatus. In order to decrease the viscosity of the substance the two-component atomizers were warmed.The following amounts of air were set:Test groupCompressed air [m³/h]Blast air [m³/h]Exhaused air [m³/h]0-3.0 ± 0.32.7 ± 0.311.5 ± 0.31.5 ± 0.32.7 ± 0.321.5 ± 0.31.5 ± 0.32.7 ± 0.331.5 ± 0.31.5 ± 0.32.7 ± 0.3Test groupSubstance flow atomizer [ml/h]Atomizer temperature [° C]10.238.9 – 41.221.2 – 4.038.7 – 39.9320 - 3538.2 – 39.9Head-nose exposure systemThe head - nose exposure technique was preferably selected for this inhalation study to minimize fur contamination of the animals with the substance, which cannot be avoided during whole-body exposure. Fur contamination may lead to an additional dermal- and oral uptake (animals preen as their fur becomes contaminited). Thus an estimation of a nominal dose, taken up by the animals and its correlation to a toxic effect becomes more difficult. Furthermore, by using the dynamic mode of operation with a Iow-volume chamber, the equilibrium characteristic of this exposure technique is favorable: t99 (the time to reach 99% of the final target concentraiion) is shorter as compared with whole - body chambers with a higher chamber volume.The aerosol was generated inside an aerodynamic exposure appalatus (INA 20; volume V ~ 55 I, BASF Aktiengesellschaft). The apparatus consists of a cylindrical inhalation chamber of stainless steel sheeting and coneshaped outlets and inlets. The rats were restrained in exposure tubes (glass tubes), their snouts projected into the inhalation chamber and they thus inhaled the aerosol. The apparatus was also connected to an exhaust air system. The exhaust air system was set lower than the supply air system (positive pressure). This ensured that the aerosol in the breathing zones of the animals was not diluted by laboratory air.In order to accustom the animals to the exposure conditions they were exposed to supply air in head-only exposure systems under comparable flow conditions on 5 days before the exposure period ( preflow period) . Then alI test groups were exposed for 6 hours on workdays over a time period of 28 days (285 days test). The number of exposures was 20.The animals did not have access to water or feed during the exposure.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

28 days

Frequency of treatment:

6 hrs/day, 5 days/week

Dose / conc.:

0.02 mg/L air (analytical)

Dose / conc.:

0.1 mg/L air (analytical)

Dose / conc.:

0.5 mg/L air (analytical)

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

Post-exposure period: none

Positive control:

None

Observations and examinations performed and frequency:

Body weight data
The body weight of the animals was checked at the beginning of the prefolw, one day before beginning of the exposure peride and then once a week alwasy at the same time of the day.Clinical signs and findings:The bahaviour and state of health of the test animals were checked on workdays at least 3 times on exposure days and one during the preflow period and the post observation days.
Mortality: A check for dead animals was made daily.
Neurofunctional tests: The evaluation on neurofunction was performed on all animals before the beginning of exposure on study days 0, day 1, 8, 14, and 27 (last exposure).The examination was performed using a functional observational battery which includes various parameters of sensoric and motoric function as follows:tremors, convulsions, piloerection, lacrimation, secretion of pigmented tears, salivation, pupil size, diarrhea, vocalization, paresis, paralysis, ataxia, general appearance, body tone, posture, animal body (appearance), locomotor activity, respiration, urination, skin color, righting reflex, behavior, grip strenght, pupillary reflex, winking reflex, vision, audition, olfaction, sensitivity of the body surface, pain perception (hot plate test), , tail pinch, toe pinch, visual placing response, miscellaneous.Clinical chemistry and HematologyBlood was taken from the retroorbital venous plexus in the morning from non-fasted, not anesthetized animals. The blood samplings and the subsequent analysis of blood and serum was carried out in a randomized sequence. The assays were performed under internal lab quality conditions with commercial reference controls to assure reliable test results.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

Statistical relevance was established using methods of ANOVA and DUNNETT-KRUSKAL-WALLIS test, MANN-WHITNEY U test

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

During the whole study period the animals of test groups 0 to 2 showed no abnormal clinical signs and findings.Reddish crusts on nasal edges (blood test positive) were detected in the animals of test group 3 after exposure during the second half of exposure period (males on days 21 to 23, 26 and 27, females on days 14 to 16, 20 to 23, 26 and 27).

Mortality:

no mortality observed

Description (incidence):

No deaths were recorded throughout the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weight and body weight change of male and female animals from alI test groups showed no statistically significant difference when compared to the control group

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No substance-induced changes were observed in the hematological parameters or in clotting analysis of both sexes. There is a statistically significant inter-group difference between the red blood cells of the male animals of the control and those of test group 2. This deviation is marginal, inconsistent, when compared with the other sex, and lacking dosage-relationship. Accordingly, this finding is considered to be of no toxicological significance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No substance-induced changes were observed in the enzyme activities or the blood chemistry parameters of both sexes.

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

The statistjcally significant increased grip strength of forelimbs in males of test group 2 on day 14 and decreased grip strength of hind limbs in females of test group 3 on day 1 were judged to be not substance related because there was no concentration or time dependence. AlI other parameters examined during neurofunctional testing did not show any difference between animals exposed to the test substance and controls. Therefore functional observational battery did not reveal any substance-induced neurofunctional impairment in the treated groups when compared to control.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Local effects were characterized histopathologically by focal inflammatory changes in the submucosa of the larynx region, which seems to be the most sensitive part of the respiratory tract after aerosol exposure to Triethanolamine. There was a tendency to concentration dependent increase in incidence and severity of the lesion from mid to high concentration in both sexes. The effect was found in females to a lesser extent. In this sex 0.02 ng/L did not cause larynx irritation anymore. In male animals however minimal to slight effects were observed in the low concentration similar to the mid concentration. Because of this simiIar grade of the lesion it is concluded that just below 0.02 ng/L no irritation should be present anymore

Key result

Dose descriptor:

NOAEC

Remarks:

systemic effects

Effect level:

500 mg/m³ air

Sex:

male/female

Basis for effect level:

other: no effects at highest concentration tested

Key result

Dose descriptor:

NOAEC

Remarks:

local effects

Effect level:

20 mg/m³ air

Sex:

female

Basis for effect level:

other: irritation of the upper respiratory tract

Key result

Dose descriptor:

NOAEC

Remarks:

local effects

Effect level:

<= 20 mg/m³ air

Sex:

male

Basis for effect level:

other: irritation of the upper respiratory tract, minimal to moderate focal inflammatory change in the submucosa of the larynx

Key result

Dose descriptor:

BMCL05

Remarks:

Local effects

Effect level:

14.1 mg/m³ air

Based on:

test mat.

Sex:

female

Basis for effect level:

other: irritation of the upper respiratory tract

Key result

Dose descriptor:

BMCL05

Remarks:

Local effects

Effect level:

14.8 mg/m³ air

Based on:

test mat.

Sex:

male

Basis for effect level:

other: irritation of the upper respiratory tract, minimal to moderate focal inflammatory change in the submucosa of the larynx

Key result

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

500 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD TG 412

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July 17 - September 05 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Principles of method if other than guideline:

Groups of 10 Wistar rats/sex were head-nose exposed to clean air (control ) and target concentrations of 0.02; 0.1 and 0.5 mg/L Triethanolamine (test groups). The substance was administered as a liquid aerosol during a period of 28 days (20 exposures). Based on the recommendation of OECD Guidelines for the "Testing of Chemicals", method 412, clinical, hematological, clinicochemical examinations were carried out. Furthermore, neurofunctjonal tests (functional observational battery) were performed before and 4 times at about weekly intervals during exposure period. At the end of the study 3 animals/sex and group were perfusion fixed for eventual further neuropathology which, however, was not performed because no neurotoxic findings were observed clinically and in routine histopathology of brain and sciatic nerve. The remaining 7 animals/sex and group were subjected to conventional necropsy. 7 animals per sex of the high concentration group and of the controls were subjected to hi stopathol ogi cal examination, the scope of organs of OECD 412 being extended by brain and peripheral nerve.

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Dr. K. Thomae GmbH, Biberach- Age at study initiation: 7 weeks- mean weight at study initiation: males: 239; females 168 g- Fasting period before study: none- Housing: individually- Diet: KLIBA rat/mouse/hamster Iaboratory diet 24-343-4 10 mm pellets, KlingentalmuehIe AG, Kaiseraugst, Switzerland ad libitum- Water: tap water ad libitumENVIRONMENTAL CONDITIONS- Temperature (°C): 20-24- Humidity (%): 30-70- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation

Type of inhalation exposure:

nose/head only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

Generation of an inhalation atmosphere: The substance to be tested was supplied to a two-component atomizer by means of a continuous infusion pump and was atomized with compressed air. Having passed a glass separator, the liquid aerosol was diluted with conditioned blast air which was conducted via a glass bottle filled with bidist. water for humidification and was supplied to the exposure apparatus. In order to decrease the viscosity of the substance the two-component atomizers were warmed. The following amounts of air were set:
Test group Compressed air [m³/h] Blast air [m³/h] Exhaused air [m³/h]0-3.0 ± 0.32.7 ± 0.311.5 ± 0.3 1.5 ± 0.32.7 ± 0.321.5 ± 0.3 1.5 ± 0.32.7 ± 0.331.5 ± 0.3 1.5 ± 0.32.7 ± 0.3
Test group Substance flow atomizer [mL/h] Atomizer temperature [° C]10.2 38.9 – 41.221.2 – 4.0 38.7 – 39.9320 - 35 38.2 – 39.9
Generation of an inhalation atmosphere: The substance to be tested was supplied to a two-component atomizer by means of a continuous infusion pump and was atomized with compressed air. Having passed a glass separator, the liquid aerosol was diluted with conditioned blast air which was conducted via a glass bottle filled with bidist. water for humidification and was supplied to the exposure apparatus. In order to decrease the viscosity of the substance the two-component atomizers were warmed.The following amounts of air were set:Test groupCompressed air [m³/h]Blast air [m³/h]Exhaused air [m³/h]0-3.0 ± 0.32.7 ± 0.311.5 ± 0.31.5 ± 0.32.7 ± 0.321.5 ± 0.31.5 ± 0.32.7 ± 0.331.5 ± 0.31.5 ± 0.32.7 ± 0.3Test groupSubstance flow atomizer [ml/h]Atomizer temperature [° C]10.238.9 – 41.221.2 – 4.038.7 – 39.9320 - 3538.2 – 39.9Head-nose exposure systemThe head - nose exposure technique was preferably selected for this inhalation study to minimize fur contamination of the animals with the substance, which cannot be avoided during whole-body exposure. Fur contamination may lead to an additional dermal- and oral uptake (animals preen as their fur becomes contaminited). Thus an estimation of a nominal dose, taken up by the animals and its correlation to a toxic effect becomes more difficult. Furthermore, by using the dynamic mode of operation with a Iow-volume chamber, the equilibrium characteristic of this exposure technique is favorable: t99 (the time to reach 99% of the final target concentraiion) is shorter as compared with whole - body chambers with a higher chamber volume.The aerosol was generated inside an aerodynamic exposure appalatus (INA 20; volume V ~ 55 I, BASF Aktiengesellschaft). The apparatus consists of a cylindrical inhalation chamber of stainless steel sheeting and coneshaped outlets and inlets. The rats were restrained in exposure tubes (glass tubes), their snouts projected into the inhalation chamber and they thus inhaled the aerosol. The apparatus was also connected to an exhaust air system. The exhaust air system was set lower than the supply air system (positive pressure). This ensured that the aerosol in the breathing zones of the animals was not diluted by laboratory air.In order to accustom the animals to the exposure conditions they were exposed to supply air in head-only exposure systems under comparable flow conditions on 5 days before the exposure period ( preflow period) . Then alI test groups were exposed for 6 hours on workdays over a time period of 28 days (285 days test). The number of exposures was 20.The animals did not have access to water or feed during the exposure.

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

28 days

Frequency of treatment:

6 hrs/day, 5 days/week

Dose / conc.:

0.02 mg/L air (analytical)

Dose / conc.:

0.1 mg/L air (analytical)

Dose / conc.:

0.5 mg/L air (analytical)

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

Post-exposure period: none

Positive control:

None

Observations and examinations performed and frequency:

Body weight data
The body weight of the animals was checked at the beginning of the prefolw, one day before beginning of the exposure peride and then once a week alwasy at the same time of the day.Clinical signs and findings:The bahaviour and state of health of the test animals were checked on workdays at least 3 times on exposure days and one during the preflow period and the post observation days.
Mortality: A check for dead animals was made daily.
Neurofunctional tests: The evaluation on neurofunction was performed on all animals before the beginning of exposure on study days 0, day 1, 8, 14, and 27 (last exposure).The examination was performed using a functional observational battery which includes various parameters of sensoric and motoric function as follows:tremors, convulsions, piloerection, lacrimation, secretion of pigmented tears, salivation, pupil size, diarrhea, vocalization, paresis, paralysis, ataxia, general appearance, body tone, posture, animal body (appearance), locomotor activity, respiration, urination, skin color, righting reflex, behavior, grip strenght, pupillary reflex, winking reflex, vision, audition, olfaction, sensitivity of the body surface, pain perception (hot plate test), , tail pinch, toe pinch, visual placing response, miscellaneous.Clinical chemistry and HematologyBlood was taken from the retroorbital venous plexus in the morning from non-fasted, not anesthetized animals. The blood samplings and the subsequent analysis of blood and serum was carried out in a randomized sequence. The assays were performed under internal lab quality conditions with commercial reference controls to assure reliable test results.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

Statistical relevance was established using methods of ANOVA and DUNNETT-KRUSKAL-WALLIS test, MANN-WHITNEY U test

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

During the whole study period the animals of test groups 0 to 2 showed no abnormal clinical signs and findings.Reddish crusts on nasal edges (blood test positive) were detected in the animals of test group 3 after exposure during the second half of exposure period (males on days 21 to 23, 26 and 27, females on days 14 to 16, 20 to 23, 26 and 27).

Mortality:

no mortality observed

Description (incidence):

No deaths were recorded throughout the study.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weight and body weight change of male and female animals from alI test groups showed no statistically significant difference when compared to the control group

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No substance-induced changes were observed in the hematological parameters or in clotting analysis of both sexes. There is a statistically significant inter-group difference between the red blood cells of the male animals of the control and those of test group 2. This deviation is marginal, inconsistent, when compared with the other sex, and lacking dosage-relationship. Accordingly, this finding is considered to be of no toxicological significance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No substance-induced changes were observed in the enzyme activities or the blood chemistry parameters of both sexes.

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

The statistjcally significant increased grip strength of forelimbs in males of test group 2 on day 14 and decreased grip strength of hind limbs in females of test group 3 on day 1 were judged to be not substance related because there was no concentration or time dependence. AlI other parameters examined during neurofunctional testing did not show any difference between animals exposed to the test substance and controls. Therefore functional observational battery did not reveal any substance-induced neurofunctional impairment in the treated groups when compared to control.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Local effects were characterized histopathologically by focal inflammatory changes in the submucosa of the larynx region, which seems to be the most sensitive part of the respiratory tract after aerosol exposure to Triethanolamine. There was a tendency to concentration dependent increase in incidence and severity of the lesion from mid to high concentration in both sexes. The effect was found in females to a lesser extent. In this sex 0.02 ng/L did not cause larynx irritation anymore. In male animals however minimal to slight effects were observed in the low concentration similar to the mid concentration. Because of this simiIar grade of the lesion it is concluded that just below 0.02 ng/L no irritation should be present anymore

Key result

Dose descriptor:

NOAEC

Remarks:

systemic effects

Effect level:

500 mg/m³ air

Sex:

male/female

Basis for effect level:

other: no effects at highest concentration tested

Key result

Dose descriptor:

NOAEC

Remarks:

local effects

Effect level:

20 mg/m³ air

Sex:

female

Basis for effect level:

other: irritation of the upper respiratory tract

Key result

Dose descriptor:

NOAEC

Remarks:

local effects

Effect level:

<= 20 mg/m³ air

Sex:

male

Basis for effect level:

other: irritation of the upper respiratory tract, minimal to moderate focal inflammatory change in the submucosa of the larynx

Key result

Dose descriptor:

BMCL05

Remarks:

Local effects

Effect level:

14.1 mg/m³ air

Based on:

test mat.

Sex:

female

Basis for effect level:

other: irritation of the upper respiratory tract

Key result

Dose descriptor:

BMCL05

Remarks:

Local effects

Effect level:

14.8 mg/m³ air

Based on:

test mat.

Sex:

male

Basis for effect level:

other: irritation of the upper respiratory tract, minimal to moderate focal inflammatory change in the submucosa of the larynx

Key result

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEC

20 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD TG 412

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Simonsen Laboratories, Gilroy, CA
- Age at study initiation: 39 to 42 days
- Weight at study initiation: 59.2 - 85.4 g (males), 57.0 - 79.3 g (females)
- Fasting period before study: No
- Housing: Individual in polycarbonate cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 - 24
- Humidity (%): 35 - 65
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

not specified

Vehicle:

acetone

Details on exposure:

TEST SITE
- Area of exposure: an area extending from the animal's mid-back to dorsal interscapular region
- Time intervals for shavings or clipplings: at least 24 hours prior to initial dose, and once weekly thereafter

REMOVAL OF TEST SUBSTANCE
No data

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0, 1.0, 0.5, 0.25, 0.125 and 0 g/kg bw
- Concentration (if solution): 1120, 560, 280, 140, 70 and 0 mg/mL
- Constant volume or concentration used: yes

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

5 days per week

Dose / conc.:

125 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

Dose / conc.:

500 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

20; 10 special study animals (designated for periodic urinalysis, hematology, and clinical chemistry determinations), 10 base study animals (subject to the collection of clinical observations data, sperm morphology and vaginal cytology evaluations, necropsy with gross examination and tissue collection, and histopathologic examination).

Control animals:

yes, concurrent vehicle

Details on study design:

Post-exposure period: no

Positive control:

None

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly


DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Daily


BODY WEIGHT: Yes
- Time schedule for examinations: Weekly


FOOD CONSUMPTION: No


FOOD EFFICIENCY: No


WATER CONSUMPTION: No


OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: Week 11
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All
- Parameters examined were erythrocyte count (RBC), hemoglobin (HgB), hematocrit (HCT), leukocyte count (WBC), platelet count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocyte differential count, reticulocyte count, erythrocyte and platelet morphology.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Week 11
- Animals fasted: No data
- How many animals: All
- Parameters examined were sorbitol dehydrogenase (SDH), glutamic-pyruvic transaminase activity (GPT), glutamic-oxaloacetic transaminase activity (GOT), urea nitrogen (BUN), creatinine, total protein, albumin, glucose.


URINALYSIS: Yes
- Time schedule for collection of urine: week 1, week 3, week 7 and week 12
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined were specific gravity, glucose, protein, microscopic examination of sediment.


NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

The following tissues were examined: gross lesions and tissue masses (and regional lymph nodes, if possible), blood smear (if required by the pathologist), mandibular and mesentric lymph nodes, salivary gland, sternebrae, femur, or vertebrae, including marrow, thyroids, heart, esophagus, stomach (to include forestomach and glandular stomach), uterus, brain (three sections, including frontal cortex and basal ganglia, parietal cortex and thalamus, and cerebellum and pons), thymus, trachea, parathyroids, small intestine (duodenum, jejunum, ileum), cecum, colon and rectum, liver, prostate, testis, epididymis, seminal vesicle, ovaries, lungs and mainstem bronchi, nasal cavity and nasal turbinates (3), preputial or clitoral glands (paired), pancreas, spleen, kidneys, adrenals, urinary bladder, pituitary, spinal cord and sciatic nerve (if neurologie signs were present), eyes (if grossly abnormal), mammary gland (including surface skin), pharynx (if grossly abnormal), skin (lesions in dosed area, unaffected skin in dosed area, and undosed control skin).

Complete histopathologic evaluation of all tissues listed was performed on all base study rats from the 2.0 and 0 g/kg dose groups. On the basis of those findings, the skin at the site of application (both males and females) and the kidney (females only)were selected as the target tissues. These tissues were examined in rats in successively lower dose groups to a no-effect level; in addition, any gross lesions detected at necropsy were examined microscopically.

Other examinations:

Sperm morphology and vaginal cytology were evaluated in the control and the three highest dose groups.

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

The only clinical abnormalities associated with treatment with triethanolamine occurred at the site of dermal dose application. Discoloration of the skin was seen in males from all dose groups (including control); it was present in 6 males in the 0 g/kg dose group, and in all 10 base study animals in each of the dosed groups. Irritation at the site of application was seen in the three highest dosed groups (0.5, 1.0, and 2.0 g/kg), with incidence increasing and time to onset decreasing with increasing dose level. Scaliness was observed only at the 1.0 and 2.0 g/kg levels; this was observed in 1 rat in the 1.0 g/kg group (first seen on Day 13), and in 5 rats in the 2.0 g/kg group (first seen on Day 6). Crustiness at the site of application was recorded for 10 rats in the 2.0 g/kg dose group; two males from this group were also observed to have ulceration at the application site. Other clinical abnormalities occurred sporadically.
All treatment-related abnormalities in females were seen at the site of dermal dose application. Irritation was observed at the 1.0 (7 rats) and 2.0 (10 rats) g/kg dose levels; scaliness was also seen in female rats from these groups (4 animals dosed at the 1.0 g/kg level, and 5 animals dosed at the 2.0 g/kg level). Crustiness was present in 3 female rats dosed at 2.0 g/kg. Other abnormalities were determined not to be compound induced.

Mortality:

no mortality observed

Description (incidence):

All animals in all dose groups survived until scheduled termination.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

The 2.0 g/kg male dose group exhibited a pronounced depression in body weight gain; the differential weight gain, relative to control, for this group was -32.8 percent, whereas those for the other male dosed groups ranged from -4.8 to +8.4 percent. The mean weight gains for all female dosed groups were depressed when compared with that of the female control group; the differential weight gains (relative to control group weight gain) for the 0.125, 0.25, 0.5, 1.0, and 2.0 g/kg female dose groups were -13.1, -10.6, -6.0, -21.4, and -35.9 percent, respectively. The mean final body weights for both male and females dosed at 2.0 g/kg body weight were significantly decreased (p <0.01), compared with those of their respective control groups.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Hematology was performed on blood obtained from special study rats on study Day 80. Male rats dosed at 2.0 g/kg exhibited a significantly increased leukocyte count and decreased mean corpuscular volume. Significantly decreased MCV was also exhibited by the 2.0 g/kg female dose group; the hematocrit of this group was depressed as well.
Male rats in the 2.0 g/kg dose group exhibited significant increases in both relative and absolute number of segmented neutrophils and eosinophils, and a significant reduction in the relative number of lymphocytes. Female rats treated at the same dosage level (2.0 g/kg) also exhibited a significant increase in relative and absolute numbers of segmented neutrophils, as well as a decrease in relative lymphocyte count. These hematological changes in high dose rats of both sexes can be attributed to an inflammatory response resulting from dermal irritation. The mean values for all other dosed groups of both sexes were statistically similar to control.

Description (incidence and severity):

The 2.0 and 0.25 g/kg male dose groups exhibited elevated SGOT levels; the mean SGPT level of the 2.0 g/kg male dose group was also significantly higher than control. No other statistically significant changes were seen when mean data from the male dosed groups were compared with that of control. The 2.0 g/kg female dose group exhibited elevated serum urea nitrogen, albumin, SGOT, and SGPT. The only other significant change in the female group mean data was a decrease in serum sorbitol dehydrogenase in rats dosed at 0.5 and 1.0 g/kg.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

Urine, collected from special study group rats on Days 3, 16, 44, and 86, was analyzed.
At study Day 3, mean values for all male dosed groups were statistically similar to those of the male control group. At study Day 16, the only values statistically different from control were urine protein at the 1.0 g/kg level, and the number of leukocytes at the 0.25 g/kg level. By study Day 44, there was a significant reduction in urine protein levels in males dosed at 0.5, 1.0, and 2.0 g/kg; in addition, the specific gravity of urine collected from the 2.0 g/kg male dose group was significantly increased over control. At the final urinalysis (Day 86), the specific gravity of urine from rats in the 2.0 g/kg dose group was again significantly elevated, and the urine protein levels for the 0.5, 1.0, and 2.0 g/kg male dose groups were significantly lower than control. The only other parameter statistically different from control was increased urine volume in males dosed at 0.5 g/kg.
For female rats, there was decreased urine protein concentration at the 0.25 g/kg level and above at the Day 3 urinalysis; the number of crystals found in the urine of rats in the 2.0 g/kg dose group was also significantly greater than control. Increased number of crystals was also present, in females dosed at 1.0 and 2.0 g/kg, on Day 16. Beginning on Day 16, and continuing through Days 44 and 86, the specific gravity of urine from female rats from these two highest dose groups was significantly greater than that of control females. The only other statistically significant changes observed at Day 44 were increased urine protein concentration in females dosed at 0.5 g/kg, and increased number of crystals in females dosed at 2.0 g/kg. At Day 86, in addition to the increased urine specific gravity in the two highest dose groups, the 2.0 g/kg females exhibited an increase in urine glucose concentration.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

The weights of the brain, right kidney, spleen, thymus, liver, lung, heart, right and left testes, and right and left epididymides were recorded at the scheduled necropsy of base study animals.
The mean brain to body weight values for the 2.0 g/kg male and female dose groups were significantly increased relative to control. In the absence of statistical changes in the absolute brain weight for these groups, this increase in relative weight appears to be the result of depressed final body weights in rats dosed at 2.0 g/kg.
Statistically significant increases in mean right kidney absolute weights and weights relative to brain weight occurred in both males and females dosed at 1.0 and 2.0 g/kg. The right kidney to body weight ratios of the 0.25, 0.5, 1.0, and 2.0 g/kg male dose groups and the 1.0 and 2.0 g/kg female dose groups were also significantly increased over that of control.
The mean spleen absolute weight and spleen weight relative to brain weight of the 2.0 g/kg female dose group were significantly decreased, relative to control values. There was a significant increase in the mean spleen to body weight ratio in male rats dosed at 1.0 and 2.0 g/kg. Thymus absolute weight and weight relative to brain weight were increased in males dosed at 2.0 g/kg body weight.
Mean liver to body weight ratios for the 0.5 and 1.0 g/kg male dose groups were significantly elevated over that of control. No statistically significant changes were observed in mean liver weight values for the female dose groups.
The mean lung absolute weight and lung to brain weight ratio of the 2.0 g/kg male dose group were significantly decreased, relative to control. The mean lung to brain weight of the 0.5 g/kg male dose group was also significantly decreased. The only statistically significant change in mean lung weight values for female rats was increased lung to brain weight ratio at the 0.125 g/kg level.
No statistically significant changes were seen in mean heart weight data for male and female rats.
For both testes, there was a significant increase in weight relative to body weight in the 2.0 g/kg male dose group; this was a result of decreased body weight at necropsy.
No statistical changes were observed for mean left epididymis weight values. The 2.0 g/kg male dose group exhibited decreased right epididymis absolute weight, and increased weight relative to body weight. Right epididymis to body and to brain weight ratios were increased in males dosed at 0.25 g/kg.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Gross lesions were common on the skin of high dose rats. The compound-related lesion seen at the application site was dermal crust. Yellow skin was seen in the lumbar region of treated and control animals. This lesion had no corresponding microscopic lesion and is attributed to application of the vehicle (acetone). Other gross lesions were considered incidental, spontaneous lesions.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The compound-related skin lesions were chronic-active inflammation and acanthosis, and were seen in 2.0, 1.0, 0.5, and 0.25 g/kg male rats and in 2.0, 1.0, and 0.5 g/kg female rats. The chronic-active inflammation contained epidermal and dermal components. The epidermal component was characterized by acanthotic, hyperkeratotic, focally parakeratotic epidermis that occasionally contained rete pegs; these changes were graded separately as acanthosis. Severely affected epidermis contained focal hemorrhage, fibrin and/or mineral deposits, bacterial colonies, serum pockets, pustules, erosions and/or ulcers. The dermal component in severely affected rats was characterized bydermal fibrosis, neocapillarization, minimally distorted adnexal organs, and variably severe mixed inflammatory infiltrates consisting of histiocytes, lymphocytes, neutrophils and eosinophils. Although only the most severe grades of chronic-active inflammation contained all of the previously listed epidermal and dermal features, the lesion was regarded as a continuum, and, therefore, the morphology "chronic-active inflammation" was used in all dose groups. Less severe lesions of chronic-active inflammation had reduced acanthosis, and lacked one or more of the following components: epidermal hemorrhage, fibrin and/or mineral deposits, bacterial colonies, serum pockets, pustules, erosions and/or ulcers. Additionally, less severe lesions contained fewer of the dermal features of the most severe lesion. In minimal inflammatory lesions the epidermal component was predominant. Where only the term acanthosis was used, the least severe lesion contained 2-3 layers of epidermal cells above the basilar layer. Occasionally, there was minimal to mild dermal fibrosis associated with the acanthosis. In males and females there was a dose dependent reduction in severity and incidence of skin lesions from high to no-effect dose.
Non-compound related microscopic lesions were seen in the kidney, liver, lung, prostate, ovary, preputial gland, clitoral gland, eye, Zymbal's gland, nose/nasal cavity; and mediastinal lymph node.

Key result

Dose descriptor:

NOAEL

Remarks:

local effects

Effect level:

125 mg/kg bw/day

Sex:

male

Basis for effect level:

other: chronic-active inflammation and acanthosis at the site of application

Key result

Dose descriptor:

NOAEL

Remarks:

local effects

Effect level:

250 mg/kg bw/day

Sex:

female

Basis for effect level:

other: chronic-active inflammation and acanthosis at the site of application

Key result

Dose descriptor:

NOAEL

Remarks:

systemic effects

Effect level:

125 mg/kg bw/day

Sex:

male

Basis for effect level:

other: increased relative kidney weight

Key result

Dose descriptor:

NOAEL

Remarks:

systemic effects

Effect level:

500 mg/kg bw/day

Sex:

female

Basis for effect level:

other: increased absolute and relative kidney weight

Key result

Critical effects observed:

not specified

The compound-induced skin lesions in males and females were chronic-active inflammation and acanthosis. The incidence, severity, and morphology of the lesions were similar between sexes. In males, however, the compound effect extended to one dose lower than in females. Although there was increased incidence of nephropathy from low to high dose in female rats, the severity of this lesion did not vary between dose groups, and, therefore, it was regarded as incidental. Other microscopic lesions were regarded as incidental and spontaneous or associated with retrobulbar bleeding.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

125 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

similar to OECD TG 411

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Simonsen Laboratories, Gilroy, CA
- Age at study initiation: 39 to 42 days
- Weight at study initiation: 59.2 - 85.4 g (males), 57.0 - 79.3 g (females)
- Fasting period before study: No
- Housing: Individual in polycarbonate cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 - 24
- Humidity (%): 35 - 65
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

not specified

Vehicle:

acetone

Details on exposure:

TEST SITE
- Area of exposure: an area extending from the animal's mid-back to dorsal interscapular region
- Time intervals for shavings or clipplings: at least 24 hours prior to initial dose, and once weekly thereafter

REMOVAL OF TEST SUBSTANCE
No data

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0, 1.0, 0.5, 0.25, 0.125 and 0 g/kg bw
- Concentration (if solution): 1120, 560, 280, 140, 70 and 0 mg/mL
- Constant volume or concentration used: yes

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

5 days per week

Dose / conc.:

125 mg/kg bw/day (nominal)

Dose / conc.:

250 mg/kg bw/day (nominal)

Dose / conc.:

500 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Dose / conc.:

2 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

20; 10 special study animals (designated for periodic urinalysis, hematology, and clinical chemistry determinations), 10 base study animals (subject to the collection of clinical observations data, sperm morphology and vaginal cytology evaluations, necropsy with gross examination and tissue collection, and histopathologic examination).

Control animals:

yes, concurrent vehicle

Details on study design:

Post-exposure period: no

Positive control:

None

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly


DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: Daily


BODY WEIGHT: Yes
- Time schedule for examinations: Weekly


FOOD CONSUMPTION: No


FOOD EFFICIENCY: No


WATER CONSUMPTION: No


OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: Week 11
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All
- Parameters examined were erythrocyte count (RBC), hemoglobin (HgB), hematocrit (HCT), leukocyte count (WBC), platelet count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocyte differential count, reticulocyte count, erythrocyte and platelet morphology.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Week 11
- Animals fasted: No data
- How many animals: All
- Parameters examined were sorbitol dehydrogenase (SDH), glutamic-pyruvic transaminase activity (GPT), glutamic-oxaloacetic transaminase activity (GOT), urea nitrogen (BUN), creatinine, total protein, albumin, glucose.


URINALYSIS: Yes
- Time schedule for collection of urine: week 1, week 3, week 7 and week 12
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters examined were specific gravity, glucose, protein, microscopic examination of sediment.


NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

The following tissues were examined: gross lesions and tissue masses (and regional lymph nodes, if possible), blood smear (if required by the pathologist), mandibular and mesentric lymph nodes, salivary gland, sternebrae, femur, or vertebrae, including marrow, thyroids, heart, esophagus, stomach (to include forestomach and glandular stomach), uterus, brain (three sections, including frontal cortex and basal ganglia, parietal cortex and thalamus, and cerebellum and pons), thymus, trachea, parathyroids, small intestine (duodenum, jejunum, ileum), cecum, colon and rectum, liver, prostate, testis, epididymis, seminal vesicle, ovaries, lungs and mainstem bronchi, nasal cavity and nasal turbinates (3), preputial or clitoral glands (paired), pancreas, spleen, kidneys, adrenals, urinary bladder, pituitary, spinal cord and sciatic nerve (if neurologie signs were present), eyes (if grossly abnormal), mammary gland (including surface skin), pharynx (if grossly abnormal), skin (lesions in dosed area, unaffected skin in dosed area, and undosed control skin).

Complete histopathologic evaluation of all tissues listed was performed on all base study rats from the 2.0 and 0 g/kg dose groups. On the basis of those findings, the skin at the site of application (both males and females) and the kidney (females only)were selected as the target tissues. These tissues were examined in rats in successively lower dose groups to a no-effect level; in addition, any gross lesions detected at necropsy were examined microscopically.

Other examinations:

Sperm morphology and vaginal cytology were evaluated in the control and the three highest dose groups.

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

The only clinical abnormalities associated with treatment with triethanolamine occurred at the site of dermal dose application. Discoloration of the skin was seen in males from all dose groups (including control); it was present in 6 males in the 0 g/kg dose group, and in all 10 base study animals in each of the dosed groups. Irritation at the site of application was seen in the three highest dosed groups (0.5, 1.0, and 2.0 g/kg), with incidence increasing and time to onset decreasing with increasing dose level. Scaliness was observed only at the 1.0 and 2.0 g/kg levels; this was observed in 1 rat in the 1.0 g/kg group (first seen on Day 13), and in 5 rats in the 2.0 g/kg group (first seen on Day 6). Crustiness at the site of application was recorded for 10 rats in the 2.0 g/kg dose group; two males from this group were also observed to have ulceration at the application site. Other clinical abnormalities occurred sporadically.
All treatment-related abnormalities in females were seen at the site of dermal dose application. Irritation was observed at the 1.0 (7 rats) and 2.0 (10 rats) g/kg dose levels; scaliness was also seen in female rats from these groups (4 animals dosed at the 1.0 g/kg level, and 5 animals dosed at the 2.0 g/kg level). Crustiness was present in 3 female rats dosed at 2.0 g/kg. Other abnormalities were determined not to be compound induced.

Mortality:

no mortality observed

Description (incidence):

All animals in all dose groups survived until scheduled termination.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

The 2.0 g/kg male dose group exhibited a pronounced depression in body weight gain; the differential weight gain, relative to control, for this group was -32.8 percent, whereas those for the other male dosed groups ranged from -4.8 to +8.4 percent. The mean weight gains for all female dosed groups were depressed when compared with that of the female control group; the differential weight gains (relative to control group weight gain) for the 0.125, 0.25, 0.5, 1.0, and 2.0 g/kg female dose groups were -13.1, -10.6, -6.0, -21.4, and -35.9 percent, respectively. The mean final body weights for both male and females dosed at 2.0 g/kg body weight were significantly decreased (p <0.01), compared with those of their respective control groups.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Hematology was performed on blood obtained from special study rats on study Day 80. Male rats dosed at 2.0 g/kg exhibited a significantly increased leukocyte count and decreased mean corpuscular volume. Significantly decreased MCV was also exhibited by the 2.0 g/kg female dose group; the hematocrit of this group was depressed as well.
Male rats in the 2.0 g/kg dose group exhibited significant increases in both relative and absolute number of segmented neutrophils and eosinophils, and a significant reduction in the relative number of lymphocytes. Female rats treated at the same dosage level (2.0 g/kg) also exhibited a significant increase in relative and absolute numbers of segmented neutrophils, as well as a decrease in relative lymphocyte count. These hematological changes in high dose rats of both sexes can be attributed to an inflammatory response resulting from dermal irritation. The mean values for all other dosed groups of both sexes were statistically similar to control.

Description (incidence and severity):

The 2.0 and 0.25 g/kg male dose groups exhibited elevated SGOT levels; the mean SGPT level of the 2.0 g/kg male dose group was also significantly higher than control. No other statistically significant changes were seen when mean data from the male dosed groups were compared with that of control. The 2.0 g/kg female dose group exhibited elevated serum urea nitrogen, albumin, SGOT, and SGPT. The only other significant change in the female group mean data was a decrease in serum sorbitol dehydrogenase in rats dosed at 0.5 and 1.0 g/kg.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

Urine, collected from special study group rats on Days 3, 16, 44, and 86, was analyzed.
At study Day 3, mean values for all male dosed groups were statistically similar to those of the male control group. At study Day 16, the only values statistically different from control were urine protein at the 1.0 g/kg level, and the number of leukocytes at the 0.25 g/kg level. By study Day 44, there was a significant reduction in urine protein levels in males dosed at 0.5, 1.0, and 2.0 g/kg; in addition, the specific gravity of urine collected from the 2.0 g/kg male dose group was significantly increased over control. At the final urinalysis (Day 86), the specific gravity of urine from rats in the 2.0 g/kg dose group was again significantly elevated, and the urine protein levels for the 0.5, 1.0, and 2.0 g/kg male dose groups were significantly lower than control. The only other parameter statistically different from control was increased urine volume in males dosed at 0.5 g/kg.
For female rats, there was decreased urine protein concentration at the 0.25 g/kg level and above at the Day 3 urinalysis; the number of crystals found in the urine of rats in the 2.0 g/kg dose group was also significantly greater than control. Increased number of crystals was also present, in females dosed at 1.0 and 2.0 g/kg, on Day 16. Beginning on Day 16, and continuing through Days 44 and 86, the specific gravity of urine from female rats from these two highest dose groups was significantly greater than that of control females. The only other statistically significant changes observed at Day 44 were increased urine protein concentration in females dosed at 0.5 g/kg, and increased number of crystals in females dosed at 2.0 g/kg. At Day 86, in addition to the increased urine specific gravity in the two highest dose groups, the 2.0 g/kg females exhibited an increase in urine glucose concentration.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

The weights of the brain, right kidney, spleen, thymus, liver, lung, heart, right and left testes, and right and left epididymides were recorded at the scheduled necropsy of base study animals.
The mean brain to body weight values for the 2.0 g/kg male and female dose groups were significantly increased relative to control. In the absence of statistical changes in the absolute brain weight for these groups, this increase in relative weight appears to be the result of depressed final body weights in rats dosed at 2.0 g/kg.
Statistically significant increases in mean right kidney absolute weights and weights relative to brain weight occurred in both males and females dosed at 1.0 and 2.0 g/kg. The right kidney to body weight ratios of the 0.25, 0.5, 1.0, and 2.0 g/kg male dose groups and the 1.0 and 2.0 g/kg female dose groups were also significantly increased over that of control.
The mean spleen absolute weight and spleen weight relative to brain weight of the 2.0 g/kg female dose group were significantly decreased, relative to control values. There was a significant increase in the mean spleen to body weight ratio in male rats dosed at 1.0 and 2.0 g/kg. Thymus absolute weight and weight relative to brain weight were increased in males dosed at 2.0 g/kg body weight.
Mean liver to body weight ratios for the 0.5 and 1.0 g/kg male dose groups were significantly elevated over that of control. No statistically significant changes were observed in mean liver weight values for the female dose groups.
The mean lung absolute weight and lung to brain weight ratio of the 2.0 g/kg male dose group were significantly decreased, relative to control. The mean lung to brain weight of the 0.5 g/kg male dose group was also significantly decreased. The only statistically significant change in mean lung weight values for female rats was increased lung to brain weight ratio at the 0.125 g/kg level.
No statistically significant changes were seen in mean heart weight data for male and female rats.
For both testes, there was a significant increase in weight relative to body weight in the 2.0 g/kg male dose group; this was a result of decreased body weight at necropsy.
No statistical changes were observed for mean left epididymis weight values. The 2.0 g/kg male dose group exhibited decreased right epididymis absolute weight, and increased weight relative to body weight. Right epididymis to body and to brain weight ratios were increased in males dosed at 0.25 g/kg.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Gross lesions were common on the skin of high dose rats. The compound-related lesion seen at the application site was dermal crust. Yellow skin was seen in the lumbar region of treated and control animals. This lesion had no corresponding microscopic lesion and is attributed to application of the vehicle (acetone). Other gross lesions were considered incidental, spontaneous lesions.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The compound-related skin lesions were chronic-active inflammation and acanthosis, and were seen in 2.0, 1.0, 0.5, and 0.25 g/kg male rats and in 2.0, 1.0, and 0.5 g/kg female rats. The chronic-active inflammation contained epidermal and dermal components. The epidermal component was characterized by acanthotic, hyperkeratotic, focally parakeratotic epidermis that occasionally contained rete pegs; these changes were graded separately as acanthosis. Severely affected epidermis contained focal hemorrhage, fibrin and/or mineral deposits, bacterial colonies, serum pockets, pustules, erosions and/or ulcers. The dermal component in severely affected rats was characterized bydermal fibrosis, neocapillarization, minimally distorted adnexal organs, and variably severe mixed inflammatory infiltrates consisting of histiocytes, lymphocytes, neutrophils and eosinophils. Although only the most severe grades of chronic-active inflammation contained all of the previously listed epidermal and dermal features, the lesion was regarded as a continuum, and, therefore, the morphology "chronic-active inflammation" was used in all dose groups. Less severe lesions of chronic-active inflammation had reduced acanthosis, and lacked one or more of the following components: epidermal hemorrhage, fibrin and/or mineral deposits, bacterial colonies, serum pockets, pustules, erosions and/or ulcers. Additionally, less severe lesions contained fewer of the dermal features of the most severe lesion. In minimal inflammatory lesions the epidermal component was predominant. Where only the term acanthosis was used, the least severe lesion contained 2-3 layers of epidermal cells above the basilar layer. Occasionally, there was minimal to mild dermal fibrosis associated with the acanthosis. In males and females there was a dose dependent reduction in severity and incidence of skin lesions from high to no-effect dose.
Non-compound related microscopic lesions were seen in the kidney, liver, lung, prostate, ovary, preputial gland, clitoral gland, eye, Zymbal's gland, nose/nasal cavity; and mediastinal lymph node.

Key result

Dose descriptor:

NOAEL

Remarks:

local effects

Effect level:

125 mg/kg bw/day

Sex:

male

Basis for effect level:

other: chronic-active inflammation and acanthosis at the site of application

Key result

Dose descriptor:

NOAEL

Remarks:

local effects

Effect level:

250 mg/kg bw/day

Sex:

female

Basis for effect level:

other: chronic-active inflammation and acanthosis at the site of application

Key result

Dose descriptor:

NOAEL

Remarks:

systemic effects

Effect level:

125 mg/kg bw/day

Sex:

male

Basis for effect level:

other: increased relative kidney weight

Key result

Dose descriptor:

NOAEL

Remarks:

systemic effects

Effect level:

500 mg/kg bw/day

Sex:

female

Basis for effect level:

other: increased absolute and relative kidney weight

Key result

Critical effects observed:

not specified

The compound-induced skin lesions in males and females were chronic-active inflammation and acanthosis. The incidence, severity, and morphology of the lesions were similar between sexes. In males, however, the compound effect extended to one dose lower than in females. Although there was increased incidence of nephropathy from low to high dose in female rats, the severity of this lesion did not vary between dose groups, and, therefore, it was regarded as incidental. Other microscopic lesions were regarded as incidental and spontaneous or associated with retrobulbar bleeding.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

Study duration:

subchronic

Species:

rat

Quality of whole database:

similar to OECD TG 411

Additional information

Oral

In a sub-chronic oral toxicity study, 20 Cox CD rats/sex/dose were exposed to 0, 250, 500 or 1000 mg/kg bw/day in the diet for 91 days (EPA, 1989b). Increased feed efficiency was observed in females of the mid-dose group. No significant differences between groups were observed in hematology and organ weights. Gross pathologic and histopathologic examination did not reveal any treatment-related effects. Thus, the NOAEL was established to be 1000 mg/kg bw/day, the highest dose tested.

Dermal

In a sub-chronic dermal toxicity study, Fischer rats were treated with 0, 125, 250, 500, 1000 or 2000 mg/kg bw/day TEA on the skin, 5 days/week for 90 days (13 weeks). 20 animals/sex/dose were exposed, of which 10 "special" animals were used for periodic urinalysis, hematology, and clinical chemistry determinations; and 10 "base" animals were used for collection of clinical observation data, sperm morphology and vaginal cytology evaluations, necropsy with gross examination and tissue collection, and histopathological examination (Battelle, 1987a). No mortality was observed. Topical application of 2000 mg/kg bw resulted in a significant decrease in body weight gain, and grossly visible crusts at the site of application were noted in males and females administered 1000 or 2000 mg/kg bw. Hematologic changes were consistent with the presence of skin inflammation in rats in the 2000 mg/kg bw groups, and clinical chemistry findings of very mild but generally dose-related increases in serum alanine and aspartate aminotransferase activities were suggestive of liver injury. However, sorbitol dehydrogenase activity, which is generally considered to be a better gauge of liver damage, was not increased, and histopathology revealed no evidence of hepatic injury. Aspartate aminotransferase has a wider tissue distribution than sorbitol dehydrogenase, and increased serum activity could be related to minor injury at another site, such as the muscle, rather than to hepatotoxicity. Additionally, some compounds can cause increases in alanine aminotransferase activity in the liver or serum without causing hepatic injury.

Kidney weights increased with increasing dose in male and female rats. Dosed males had decreased urinary protein excretion which likely reflected a change in renal function or an increase in protein reabsorption, as serum protein concentrations were not affected. Although these findings suggest the possibility of protein droplet accumulation or some other form of renal dysfunction or injury, no evidence of hyaline droplet nephropathy or other histopathologic changes that might account for the weight changes was noted.

Lesions at the site of application ranged from no discernable change, through minimal to mild epidermal thickening (acanthosis), to chronic active inflammation, erosion, and ulceration. The dermis was also thickened with inflammation and fibrosis at the higher doses. There was no histological evidence to suggest the development of skin sensitisation or contact dermatitis. NOAELs for local effects were determined to be 125 and 250 mg/kg bw/day for males and females, respectively. The NOAELs for systemic effects were established to be 125 and 500 mg/kg bw/day for males and females, respectively, based on kidney effects.

In a sub-chronic dermal toxicity study using an identical experimental set-up as in the study with rats, mice were exposed to 0, 250, 500, 1000, 2000 or 4000 mg/kg bw/day TEA on the skin ( (Battelle, 1987b). Findings were similar to those in the rat study. All mice survived to the end of the study. Clinical findings were observed only in mice of the 4000 mg/kg bw group and included scaliness, irritation, and discoloration at the application site for males and females, and skin erosion in one male. The absolute kidney and liver weights of males and females administered 4000 mg/kg bw were greater than those of the vehicle controls; relative kidney weights of males administered 1000 mg/kg bw or greater and females in all dosed groups were also greater than those of the vehicle controls. Microscopic examination of the skin of dosed mice indicated acanthosis and inflammation at the site of application. Acanthosis occurred in all dosed groups and in one vehicle control female; the severity increased with increasing dose in males and females. Inflammation was only observed in males and females in the 4000 mg/kg bw groups and in one female in the 2000 mg/kg bw group. The NOAEL for local effects was determined to be ≤ 250 mg/kg bw/day. NOAEL's for systemic effects were established to be 1000 and ≤ 250 mg/kg bw/day for males and females, respectively, based on kidney effects.

Inhalation

Repeated inhalation toxicity was investigated in a sub-acute 28 -day study performed according to OECD TG 412 (BASF SE, 1993) and under GLP conditions, in which Wistar rats (10/sex/dose) were exposed head/nose only to 0, 0.02, 0.1 or 0.5 mg/L TEA for 6 hours/day and 5 days/week. No mortality was observed. No statistically significant differences between groups were observed in body weight, haematology and clinical chemistry. Differences in grip strength were judged not substance-related because of a lack of concentration- or time-related effect. No other abnormalities were observed during neurofunctional testing. A significant difference in red blood cells was observed in males of the mid-dose group compared to controls, but since this deviation was marginal, not observed in females, and not dose-related, this finding was considered of no toxicological significance. Local effects were characterized histopathologically by focal inflammatory changes in the submucosa of the larynx, with a concentration-dependent tendency in incidence and severity. No such effects were observed in females of the low dose, whereas minimal to slight effects were seen in males at this dose. Therefore, 0.02 mg/L was considered to be the NOAEC for local effects in females and the LOAEC for males. Since no systemic effects were observed, the systemic NOAEC was established to be 0.5 mg/L, the highest dose tested.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The substance is not considered to be classified for repeated dose toxicity according to Regulation (EC) No 1272/2008 (CLP), as amended for the tenth time in Regulation (EU) No 2017/776.