Alcohols, C10-16, ethoxylated, sulfates, mono(hydroxyethyl)ammonium salts

Endpoint summary

Description of key information

The assessment is based on the data currently available. New studies, based on the category review and the final decisions issued for some of the category substances, which are also relevant for this assessment, are currently being conducted. The hazard assessment with respect to repeated dose toxicity will be updated once all ongoing studies have been finalised.

No data are available for the target substance Alcohols, C10-16, ethoxylated (1-2.5 EO), sulfates, monoeth. salts (CAS 157627-92-4). Therefore, read-across from structural analogue substances has been applied.

RDT (oral): For the whole category of alcohol ethoxysulfates (AES) an oral NOAEL of 300 mg/kg bw/day was established.

Read-across from structural analogue source substances Fatty alcohol C12-C14, ethoxylated (2 EO) sulphated sodium salt (CAS 68891-38-3).

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Urinalysis and neurobehaviour were not examined.

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

According to Guideline.

Route of administration:

oral: gavage

Vehicle:

water

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

90 days

Frequency of treatment:

Daily

Dose / conc.:

25 mg/kg bw/day (nominal)

Remarks:

nominal in water

Dose / conc.:

75 mg/kg bw/day (nominal)

Remarks:

nominal in water

Dose / conc.:

225 mg/kg bw/day (nominal)

Remarks:

nominal in water

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

According to Guideline.

Positive control:

Not necessary

Observations and examinations performed and frequency:

Urinalysis and neurobehaviour were not examined. Otherwise according to Guideline.

Sacrifice and pathology:

According to Guideline.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

> 225 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: No toxic effects up to and including the highest dose level tested

Key result

Critical effects observed:

no

Additional repeated dose toxicity studies supporting the key results:

CAS 68585-34-2, 1977a: NOAEL (90d) = 100 mg/kg bw/day

CAS 68585-34-2, 1977b: NOAEL (90d) >= 600 mg/kg bw/day

AES C12 Na, 1967: NOAEL (90d) > 250 mg/kg bw/day

AES C12 Na, 1991: NOEL (2y) > 250 mg/kg bw/day

Further supporting data are derived from a 2 -generation study (refer to sec. 7.8.1):

CAS 68891-38-3, 1999: NOAEL = 300 mg/kg bw/day

Conclusions:

For the systemic toxicity after repeated oral application a NOAEL of greater than 225 mg/kg bw/day can be deduced since no effects were observed at doses up to and including 225 mg/kg bw, which was the highest dose tested.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

not applicable

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: CD

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River U.K. Ltd., Margate, Kent
- Age at study initiation: 3 weeks
- Weight at study initiation: 40-50 g
- Fasting period before study: No
- Housing: Five rats per cage (suspended polypropylene cages measuring 52 x 35 x 18 cm with stainless steel mesh floors and lids)
- Diet: Complete powdered rodent diet (Spratts Laboratory Animal Diet No. 2), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 3 days, any rat that failed to gain weight at this time was discarded

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 1
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): A pre-mix containing 215,000 ppm test material was prepared each week and, from this, the different dietary concentrations were obtained by direct dilution with further quantities of diet
- Mixing appropriate amounts with (Type of food): Complete powdered rodent diet (Spratts Laboratory Animal Diet No. 2)

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks (91 days)

Frequency of treatment:

Continuous through the diet

Dose / conc.:

0.05 other: % (w/w) of test material (nominal in diet)

Remarks:

0.015% active substance

Dose / conc.:

0.5 other: % (w/w) of test material (nominal in diet)

Remarks:

0.15% active substance

Dose / conc.:

5 other: % (w/w) of test material (nominal in diet)

Remarks:

1.5% active substance

Dose / conc.:

10 other: mg/kg bw/day active substance

Remarks:

nominal in diet

Dose / conc.:

100 other: mg/kg bw/day active substance

Remarks:

nominal in diet

Dose / conc.:

1 100 other: mg/kg bw/day active substance

Remarks:

nominal in diet

No. of animals per sex per dose:

20

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: Based upon the result of a 14-day preliminary study, which indicated that the test substance incorporated into the diet at 5% w/v would result in a 10% reduction in bodyweight over the treatment period; it was also requested to examine the lower treatment levels of 1/10 and 1/100 the upper level (i.e. 0.5% w/v and 0.05% w/w).
- Rationale for animal assignment (if not random): The mean bodyweights of the male and female subgroups were adjusted by exchange of animals until they all fell within a range of 60-61 g (males) or 59-60 g (females)

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily
- Cage side observations included: Mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily
- All animals were physically examined at appropriate intervals, or otherwise sensible, signs of ill-health or reaction to treatment

BODY WEIGHT: Yes
- Time schedule for examinations: The weight of each rat was recorded on the day that treatment commenced and subsequently at weekly intervals throughout the treatment period

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- The quantity of food eaten by each cage of rats was calculated weekly, by measurements of the amount of food given, and that remaining in the food hoppers

FOOD EFFICIENCY: Yes
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes (food conversion ratios calculated on a per cage basis)

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the conclusion of the treatment period
- Anesthetic used for blood collection: No data
- Animals fasted: No data
- Collection method: Blood samples were withdrawn from the retro-orbital sinus using EDTA as anticoagulant
- How many animals: Five male and five female rats from groups 1 and 4 each (on account of findings in the female rats of group 4, the examination of hemoglobin concentration, erythrocyte count and packed cell volume was extended to similar numbers or female rats in groups 2 and 3 at week 13)
- Parameters examined: hemoglobin concentration (Hb) as cyanmethemoglobin, erythrocyte count (RBC), total leukocyte count (WBC), differential leukocyte count (WBC) and packed cell volume (PCV) as microhematocrit
- From the erythrocytic data, the mean cell hemoglobin concentration and mean cell volume were calculated for each sample

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the conclusion of the treatment period
- Animals fasted: No data
- How many animals: Five male and five female rats from groups 1 and 4 each (on account of findings in the rats of group 4, the examination of alkaline phosphatase activity and glutamate-pyruvate transaminase activity was extended to similar numbers or rats in groups 2 and 3 at week 13)
- Parameters examined: alkaline phosphatase activity (SAP) and glutamate-pyruvate transaminase activity (SGPT)

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data

Sacrifice and pathology:

SACRIFICE AND PATHOLOGY
- All rats were killed after 13 weeks of treatment by carbon dioxide inhalation; necropsy followed as soon as possible

GROSS PATHOLOGY: Yes
- A detailed necropsy was performed with opening of the cranial, thoracic and abdominal cavities; the appearance of any abnormal tissues was recorded.

HISTOPATHOLOGY: Yes
- Samples of the following tissues were taken from each rat and preserved in buffered 4% formaldehyde saline and examined microscopically: adrenal glands, brain, duodenum, heart, ileum, jejunum, kidneys, liver, lungs, lymph nodes (cervical and mesenteric), ovaries, spleen, stomach, testes and urinary bladder
- After dehydration and embedding in paraffin wax, sections of the required tissues were cut at 5 µm thickness and stained with hematoxylin and eosin. Both glandular and non-glandular areas of the stomach, and both auricular and ventricular sections of the heart, were prepared. The brain was sectioned at three levels (cerebellum, cerebral cortex and medulla).

Other examinations:

- Organ weight analysis: adrenal glands, brain, heart, kidneys, liver, ovaries, spleen, testes and thyroid glands
- The ratio of organ to bodyweight was calculated in each case (the ratio of organ weight to brain weight was also determined for the liver, kidney, spleen and heart)
- The following were not examined histopathologically but were taken from ten male and ten female rats per group and stored in fixative for possible future requirement for microscopic evaluation: aorta, colon, eye, esophagus, pituitary, prostate gland, skeletal muscle, skin, spinal cord, trachea, thyroids and uterus.
- A bone marrow smear was taken from each rat, air-dried and fixed in methanol
- The eye was preserved in Davidson’s fluid

Statistics:

The significance of any inter-group differences in organ weights, blood composition, and growth performance was assessed by analysis of variance.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

effects observed, treatment-related

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not specified

Other effects:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
- No mortality occurred
- There was no observable difference in the appearance, mood, locomotion, and fecal consistency of treated and control animals, and no animals died during the treatment period.

BODY WEIGHT AND WEIGHT GAIN:
- The bodyweight gains of both sexes in the 5.0% group were reduced from the commencement of treatment, so that their bodyweights were significantly different from the controls when analyzed at week 2 (p < 0.01, analysis of variance); at termination their bodyweights were 84% (males) and 85% (females) of those of the respective controls
-The bodyweight gains of both sexes in the 0.5% group were depressed from week 5 (males) or week 6 (females), although this reduction was statistically significant in the males only
- The growth of either sex was undisturbed in the 0.05% group

FOOD CONSUMPTION AND COMPOUND INTAKE: No information regarding statistical significance provided.
- A small reduction in food consumption occurred in both sexes receiving in the 5.0% group during the first 5 weeks of treatment; total food intakes being 93% (males) and 90% (females) of the respective controls. The food consumption of females in the 5.0% group remained depressed until week 10, but thereafter was unaffected, whereas the food intake of the males at this treatment level was unaffected from week 6 to week 13
- The food consumption of females in the 0.50% was slightly reduced over the first seven weeks of treatment, but unaffected thereafter; the food intakes of males at this treatment level, and of both sexes at the lowest level, were unaffected throughout the treatment period
- Food consumption was inconspicuous for the 0.05% group

FOOD EFFICIENCY: No information on statistical significance provided.
- The food conversion ratios of males in the 5.0% group were consistently greater than those of the controls, indicating inferior food utilization in this group
- The food conversion ratios of females at this treatment level did not demonstrate a clear pattern from week to week, but the overall (week 1-13) value also indicated slightly inferior food utilization
- Rats at the low and intermediate treatment levels showed comparable food conversion ratios to those of the controls.

HAEMATOLOGY
- Females in the 5.0% and 0.5% groups displayed erythrocytic characteristics slightly lower than in controls, statistical significance being achieved only in packed cell volume (5% dose group) and erythrocyte count (0.5% dose group)

CLINICAL CHEMISTRY
- SGPT and SAP levels were significantly elevated in both sexes in the 5.0% group
- The differences in alkaline phosphatase levels in the males were exaggerated by the contribution of one control rat which yielded a very low value (8 i.u./L; confirmed by a duplicate analysis). In a statistical analysis omitting this animal, the significance of the difference between the group mean values of the high treatment level and control groups was lower (p < 0.01)
- At the lower treatment levels, occasional animals showed elevations of SGPT or SAP, but neither consistent intergroup variations nor significant deviations from control values were evident.

ORGAN WEIGHTS
- Ratios of organ weights to bodyweight were calculated since bodyweight differences between the control, the 0.5 and 5% dose groups obscured the comparison of organ weights relative to bodyweight
- The liver weights of both sexes in the 5.0% group were significantly elevated.
- The heart weights of males in the 5.0% group were significantly reduced when analyzed in absolute terms and relative to brain weight, as also were spleen and kidney weights relative to brain weight.
- Although there were occasional deviations from the control values in the weights of various organs from males treated at the low and intermediate treatment levels when analyzed relative to bodyweight, there were no consistent changes in these organ weights when analyzed in absolute terms or relative to brain weight.

GROSS PATHOLOGY
- No treatment-related changes were seen in the macroscopic appearance of the tissues examined at necropsy

HISTOPATHOLOGY: NON-NEOPLASTIC
- No histopathological changes or variations from normal were seen that were considered to be related to treatment

Dose descriptor:

NOAEL

Effect level:

0.15 other: % in feed

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Lower body weight gain in males in final eight weeks of the study.

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Lower body weight gain in males in final eight weeks of the study.

Dose descriptor:

LOAEL

Effect level:

1.5 other: % in feed

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Lower body weight gain, small reduction in red cell parameters, elevation in serum SGPT and alkaline phosphatase activity, elevations in absolute and relative liver weights.

Dose descriptor:

LOAEL

Effect level:

1 100 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Lower body weight gain, small reduction in red cell parameters, elevation in serum SGPT and alkaline phosphatase activity, elevations in absolute and relative liver weights.

Critical effects observed:

not specified

Gonadal tissues were examined for both gross pathology and histopathology and for organ weight analysis and no treatment-related effects were detected.

Conclusions:

Since the decrease in the erythrocyte count was not dose-dependent, it was concluded that the test substance at dietary levels up to 0.5% test material (corresponding to 100 mg/kg bw/day) elicited no adverse effects.

Reference 3

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

29 Jun 1976 - 21 Feb 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

not applicable

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Wilmington, Massachusetts.
- Weight at study initiation: 112-159 g (males), 102-132 g (females)
- Housing: individually housed
- Diet: ad libitum
- Water: ad libitum

IN-LIFE DATES: From: 1976-06-29 To: 1976-10-06

Route of administration:

oral: feed

Vehicle:

other: plain diet

Details on oral exposure:

DIET PREPARATION
- Appropriate amounts of the test material were incorporated into the basal diet on a weight/weight basis and thoroughly mixed.
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Purina Laboratory Chow
- Storage temperature of food: 22-26°C

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of each freshly prepared diet were submitted to the sponsor for analysis

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

continuous

Dose / conc.:

0.1 other: % active substance in diet

Remarks:

nominal in diet

Dose / conc.:

0.5 other: % active substance in diet

Remarks:

nomonal in diet

Dose / conc.:

1 other: % active substance in diet

Remarks:

nominal in diet

Dose / conc.:

60 other: mg/kg bw/day active substance

Remarks:

nominal in diet

Dose / conc.:

300 other: mg/kg bw/day active substance

Remarks:

nominal in diet

Dose / conc.:

600 other: mg/kg bw/day active substance

Remarks:

nominal in diet

No. of animals per sex per dose:

20

Control animals:

yes, plain diet

Details on study design:

- Rationale for animal assignment (if not random): by individual, initial body weight so that a homogenous distribution of mean weights and weight ranges was obtained among groups.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

FOOD EFFICIENCY: Yes
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to treatment and at week 13
- Dose groups that were examined: some animals from all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 5 rats/sex/group at week 4 and 10 rats/sex/group at week 13
- How many animals: 5 rats/sex/group at week 4 and 10 rats/sex/group at week 13

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at sacrifice
- Animals fasted: Yes
- How many animals: 5 rats/sex/group at week 4 and 10 rats/sex/group at week 13

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes. The following organs of each rat were weighed and the organ/body weight ratios determined: heart, liver, kidneys, testes with epididymides, pituitary, adrenals, and ovaries.

HISTOPATHOLOGY: Yes. Brain with meninges, pituitary, thoracic spinal cord, thyroids, trachea, esophagus, salivary glands, eyes, tongue, lungs, heart, liver, kidneys, spleen, stomach, pancreas, lymph nodes, small and large intestine, adrenals, urinary bladder, ureters, urethra, testes, ovaries, prostate, uterus, vagina, seminal vesicles, inguinal mammary gland, skin, bone, muscle, and any unusual lesions. (5 rats/sex sacrificed at 4 weeks from the control and high-dose groups and 10 rats/sex sacrificed at 13 weeks from the control and high-dose groups. Also, kidney and liver sections from the low- and mid- dose groups from 5 rats/sex, sacrificed at 13 weeks were examined)

Statistics:

The following data from control and treated groups of the same sex were statistically compared: growth rate and total food consumption, clinical laboratory data, and terminal body weights, organ weights, and organ/body weight ratios.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

not examined

Details on results:

CLINICAL CHEMISTRY:
Differences of statistical significance noted included the following: lower than control mean glutamic-oxaloacetic transaminase values in all of the treated males at week 4 and all of the treated females at week 13; lower than control mean alkaline phoshatase values in all of the treated females at week 4; higher than control mean glucose values in the low- and high-dose females at week 13; lower than control mean albumin values in the high-dose males at week 4 and the low-dose females at week 13; a higher than control mean sodium values in the high-dose females at week 4; and higher than control mean calcium values in the high-dose females at week 4 and the low-dose females at week 13.

ORGAN WEIGHTS:
The liver weights and ratios of all of the treated groups of both sexes were slightly to moderately higher than respective control data at weeks 4, 13, and 14. These differences were generally (but not always distinctly) dose-related and statistically significant at some of the intervals. The heart weights and ratios of all of the male treated groups were slightly, (not statistically significantly) lower than those of the controls at weeks 4, 13, and 14. Microscopic examination of liver and heart sections did not confirm the presence of any treatment-related hepatic or cardiac alterations.

Dose descriptor:

NOAEL

Effect level:

>= 1 other: % in the diet

Based on:

act. ingr.

Sex:

male/female

Dose descriptor:

NOAEL

Effect level:

>= 600 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Critical effects observed:

not specified

Gonadal tissues were examined for both gross pathology and histopathology and no treatment-related effects were detected.

Conclusions:

No outstanding treatment related toxic effects were noted for the test substance administered up to 1% in the diets. Although there were statistically significant dose-related differences in absolute and relative liver and heart weights noted in test substance-treated groups compared to control animals, no histopathological correlation was found to confirm the presence of any treatment-related alterations. As such, the NOAEL for the study was determined to be >= 1% in the diet, corresponding to 600 mg/kg bw/day.

Executive summary:

This study was designed to evaluate the toxicological effects of the test substance in albino rats (20/sex/group) when administered in the diet for thirteen weeks at levels of 0.0, 0.1, 0.5, and 1.0%, corresponding to 0, 60, 300 and 600 mg/kg bw/day, respectively, active ingredient. Interim sacrifices were performed on five rats/sex/group at four weeks and ten rats/sex/group at thirteen weeks. Five rats/sex/group were maintained for one additional week and sacrificed at Week 14. The criteria evaluated for compound effects were clinical signs, mortality, body weight and food consumption, ophthalmoscopic findings, clinical chemistry and haematology, organ weights, and gross and microscopic pathology. 

No distinct effect attributed to the administration of the test substance was noted in comparisons of the clinical signs, mortality rates, ophthalmoscopic findings or haematology and blood chemistry values of the test groups to the controls.  In addition, no compound related gross or histomorphologic organ or tissue alterations were noted in any of the treated animals. 

When compared to the data of the control group, slightly lower body weight gains were noted in the mid- and high-dose male groups.  These differences were not statistically significant. 

The absolute and relative liver weights of all treated groups of both sexes were slightly to moderately higher than respective control data at Weeks 4, 13 and 14.  These differences were generally (but not always distinctly) dose-related and statistically significant at some of the intervals.  The heart weights and ratios of all of the male treated groups were slightly, (not statistically significantly) lower than those of the controls at Weeks 4, 13 and 14.  However, neither gross pathology, nor microscopic examination of liver and heart sections confirmed the presence of any treatment-related hepatic or cardiac alterations. 

Gonadal tissues were examined for both gross pathology and histopathology and no treatment-related effects were detected.

No outstanding treatment related toxic effects were noted when the test substance was administered up to 1% in the diet to albino rats. Although there were statistically significant differences in absolute and relative liver and heart weights noted in test substance-treated groups compared to control animals, and these differences were generally dose-related, no histopathological correlation was found to confirm the presence of any treatment-related alterations. As such the NOAEL was determined to be the 1% dose level, corresponding to 600 mg/kg bw/day.

Reference 4

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

Deviations:

yes

Remarks:

Only limited data are available

Dose descriptor:

NOEL

Effect level:

> 250 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: Feeding study

Dose descriptor:

NOEL

Effect level:

> 75 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: Drinking water study

Critical effects observed:

not specified

Reference 5

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment.

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

Only limited data are available

Dose descriptor:

NOAEL

Effect level:

> 250 mg/kg bw/day (nominal)

Sex:

male/female

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

300 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

The whole data base is conclusive and sufficient for assessment of the oral repeated dose toxicity potential of the target substance.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The assessment is based on the data currently available. New studies, based on the category review and the final decisions issued for some of the category substances, which are also relevant for this assessment, are currently being conducted. The hazard assessment with respect to repeated dose toxicity will be updated once all ongoing studies have been finalised.

No data on repeated dose toxicity are available for AES (C12-14; < 2.5 EO) C2H7NO (CAS 157627-92-4). Therefore, this endpoint is covered by read-across from structurally related AES. The AES reported within the category show similar structural, physico-chemical, environmental and toxicological properties. The approach of grouping different AES for the evaluation of their effects on human health and the environment was also made by the Danish EPA (2001) and HERA (2003), supporting the read-across approach between structurally related AES.

Repeated dose toxicity: oral

In the subchronic oral gavage key study according to OECD Guideline 408, the analogue substance AES (C12-14) Na (CAS 68891-38-3, no data on grade of ethoxylation) was tested for systemic toxicity at doses of 0, 25, 75, 225 mg/kg bw/day on Wistar rats (BASF, 1994b). No clinical signs of toxicity, no mortalities and no effects on any other investigated parameter (body weight, body weight gain, food consumption, water consumption, ophthalmoscopic examinations, haematology, clinical chemistry, organ weights, gross pathology and histopathology) were seen at the highest dose level. Apart from the missing systemic toxicity, local treatment-related concentration-dependent irritation to different degrees in the forestomach was seen in all test groups. Thus, a NOEL-value was not determined. Since there is no human equivalent to the rat forestomach, these effects are not considered to be relevant to human health assessment. Thus, a no adverse effect level (NOAEL) of greater than 225 mg/kg bw/day could be established.

In a supporting subchronic feeding study, four groups of 20 rats per sex were fed 0, 0.05, 0.5 and 5.0% (w/w) of the analogue compound AES (C10-16; 3 EO) Na (CAS 68585-34-2) over a period of 91 days (P&G, 1977a).  The test material contained 30% active ingredient, converting the doses to 0, 10, 100 and 1100 mg/kg bw/day active substance. There were no observable differences in the appearance, mood, locomotion, and fecal consistency of treated and control animals and no animals died during the treatment period. No statistically significant changes in food consumption occurred during the study. The body weight gains of both sexes in the 5.0% group were reduced from the commencement of treatment. Body weights were significantly different from the controls when analyzed at Week 2. At termination their body weights were 84% (males) and 85% (females) of those of the respective controls. The body weight gains of both sexes in the 0.5% group were lower from Week 5 (males) or 6 (females), although this reduction was statistically significant in the males only. Body weight gains in the 0.05% group were inconspicuous when compared to control. The food conversion ratio of males in the 5.0% group was consistently reduced, indicating inferior food utilization; however, it is not specified if this finding was statistically significant. With regard to hematology, females in the 5.0% group displayed erythrocytic characteristics slightly lower than in the controls, with statistically significant effects on packed cell volume and erythrocyte count. No hematological effects were observed in any other dose group. With regard to clinical chemistry, serum glutamate pyruvate transaminase and alkaline phosphatase levels were significantly elevated in both sexes in the 5.0% group. No further effects regarding clinical chemistry have been detected for any other dose group. The liver weights of both sexes in the 5.0% group were significantly elevated. The heart weights of both sexes in the 5.0% group were significantly reduced when analyzed in absolute terms and relative to brain weight. At necropsy, no treatment-related changes were seen in the macroscopic appearance of the tissues examined at necropsy. Other dose levels were inconspicuous at necropsy. No histopathological changes or variations that were considered treatment-related were observed in any of the tissues and organs examined. Regarding histopathology, several effects were seen in the lungs, liver and kidneys; however, they were not statistically significant and were considered not to be of toxicological significance. There were no treatment-related effects on the organ weights (both absolute and relative), or macroscopic or microscopic pathology of the gonadal tissues examined. Since the decrease in the erythrocyte count was not dose-dependent, it was concluded that the test substance at dietary levels up to and including 0.05% test material (corresponding to 100 mg/kg bw/day) elicited no adverse effects. The No-Observed-Adverse-Effect-Level (NOAEL) was therefore determined to be 100 mg/kg bw/day.

A further subchronic study conducted with the same analogue compound as above also is considered for support. In fact, this study was designed to evaluate the toxicological effects of AES (C10-16; 3 EO) Na (CAS 68585-34-2) in albino rats (20/sex/group) when administered in the diet for thirteen weeks at levels of 0.0, 0.1, 0.5, and 1.0% active ingredient (corresponding to 0, 60, 300 and 600 mg/kg bw/day) (P&G, 1977b). Interim sacrifices were performed on five rats/sex/group at four weeks and ten rats/sex/group at thirteen weeks. Five rats/sex/group were maintained on study for one additional week and sacrificed at Week 14. The criteria evaluated for compound effects were clinical signs, mortality, body weight and food consumption, ophthalmoscopic findings, clinical chemistry and haematology, organ weights, and gross and microscopic pathology. No distinct effect attributed to the administration of the test substance were noted in comparisons of the clinical signs, mortality rates, ophthalmoscopic findings or haematology and blood chemistry values of the test groups to the controls. In addition, no compound related gross or histomorphologic organ or tissue alterations were noted in any of the treated animals. When compared to the data of the control group, slightly lower body weight gains were noted in the mid- and high-dose male groups. These differences were not statistically significant. The absolute and relative liver weights of all treated groups of both sexes were slightly, to moderately higher than respective control data at Weeks 4, 13 and 14. These differences were generally (but not always distinctly) dose-related and statistically significant at some of the intervals. The heart weights and ratios of all of the male treated groups were slightly (not statistically significantly) lower than those of the controls at Weeks 4, 13 and 14. However, neither gross pathology, nor microscopic examination of liver and heart sections confirmed the presence of any treatment-related hepatic or cardiac alterations. Gonadal tissues were examined for both gross pathology and histopathology and no treatment-related effects were detected. No outstanding treatment related toxic effects were noted when the test substance was administrated up to 600 mg/kg bw/day in the diet to albino rats. Although there were statistically significant differences in absolute and relative liver and heart weights noted in test substance-treated groups compared to control animals, and these differences were generally dose-related, no histopathological correlation was found to confirm the presence of any treatment-related alterations. As such the NOEL was determined to be 600 mg/kg bw/day.

In a further supporting study, synthetic AES (C12-15; 3 EO) Na and natural AES (C12; 3 EO) Na (sodium lauryl(3EO)ethoxysulphate) were tested in a 90-day rat diet study at dose levels of 0, 40, 200, 1000 and 5000 ppm active material, corresponding to 0, 2, 10, 50 and 250 mg/kg bw/day (Walker, 1967). Health, behaviour, body weight, food intake, haematological and urinary parameters remained within normal limits at all doses. In both studies organ weight and blood chemistry effects observed were unaccompanied by any pathological changes. Based on the available information and taking into account that the study was conducted prior to the development of GLP and OECD guidelines, a NOAEL could be established at the dose level of greater than 250 mg/kg bw/day.

No unusual findings regarding systemic toxicity were noted in a 2-year chronic feeding study in rats in which AES (C12; 3EO) (lauryl(3EO)ethoxysulphate) was given at 0, 0.1 or 0.5% (corresponding to 0, 50, 250 mg/kg bw/day) in the diet (Little, 1991). The results of this study suggest that the NOEL for AES (C12; 3EO) in this 2-year chronic feeding study in rats was greater than 250 mg/kg bw/day. In another 2-year study (Little, 1991), rats were administered AES (C12; 3 EO) (lauryl(3EO)ethoxysulphate) in the drinking water at a concentration of 0.1% (equals a dose of 75 mg/kg bw/day). The only unusual finding was slight, but consistently higher water consumption by all rats receiving the test compound in their drinking water and a significant difference in the empty caecum to body weight ratio of females. Absolute organ weights were all comparable to controls and no consistent gross or histopathology was found. A NOEL greater than 75 mg/kg bw/day can be estimated on the basis of the available information.

 

Further information on subchronic toxicity can be deduced from a two-generation reproduction study with AES (C12-14, no data on grade of ethoxylation) Na (CAS 68891-38-3) (BASF, 1999). Sprague Dawley rats were dosed via the drinking water at the concentrations 0, 0.03, 0.1 and 0.3%, which corresponded to daily doses of ca. 0, 30, 100 and 300 mg/kg bw/day. There were some changes indicative of parental toxicity in the group treated with 0.3% of the test substance. Slight but significantly reduced straight line velocity (VSL) of the sperm was without any significant effects on averaged path velocity (VAP) or total motility. Moreover, in the available subchronic and chronic toxicity studies on various AES the primary sex organs of the males and females did not show evidence for treatment-related adverse effects. The observed reduced triglyceride levels (female) and increased percentage neutrophil counts (males) were slight and within the range of the historical control data. The male F0 generation showed a small but significant reduction in body weight-liver weight ratios, but the corresponding brain related liver weights and the absolute liver weights developed not in a dose dependant way. For the F1 generation where similar results were reported, no dose-response relationship was detected either. No influence on liver weight development was seen in the F2 generation. None of the groups revealed any histopathological or clinical-chemical findings, which could be attributed to hepatotoxicity. This led to the conclusion that this untypical liver weight reduction was of no toxicological relevance, additionally underlined by the absence of such effects in the studies for subchronic toxicity mentioned above. There was evidence of toxicity on pup development at this dose level that was characterised by an increase in the time taken for sexual development of the male (not significant) and female (significant) offspring. This was investigated in more detail in the developmental toxicity studies up to 1500 mg/kg bw/day and no effects were noted there. Considering all these facts the subchronic NOAEL for systemic toxicity can be set to greater than 300 mg/kg bw/day.

 

The NOAELs from the repeated oral toxicity studies were considered for the risk assessment. The respective NOAELs and LOAELs are listed in Table 1. 

 

Table 1: Dietary NOAELs and LOAELs (a.i.) for repeated oral toxicity studies of AES

Substance	Duration (weeks)	NOAEL/NOEL (mg/kg bw/day)	LOAEL (mg/kg bw/day)	Reference
AES(C12-14)Na	13	225	> 225	BASF, 1994b
AES(C12-15;3EO)Na AES(C12;3EO)Na	13	250	> 250	Walker, 1967
AES(C10-16;3EO)Na	13	100	1100	P&G, 1977b
AES(C10-16;3EO)Na	13	600	> 600	P&G, 1977c
AES(C12-14)Na	13	300	> 300	BASF, 1999
AES(C12;3EO)	104	250	> 250	Little, 1991

 

No LOAELs could be detected in the most studies, except for one study were a LOAEL was established at 1100 mg/kg bw/day (P&G, 1977b). However the dose tested exceeded the next lower dose within this study by 11 times. In addition the LOAEL is almost twice of the highest reported NOAEL. With exception of this study all NOAELs represented the highest dose level. Therefore an average of all NOAEL was chosen as basis for the risk assessment. The available oral toxicity studies provide a coherent picture on the subchronic and chronic oral toxicity of AES. Based on the described effects, the NOAEL of 300 mg/kg bw/day (BASF, 1999) representing an average of all NOAELs, was chosen for the risk assessment.

 

Influence of counter ions on repeated dose toxicity

Since monoethanolamine (MEA) in its protonated form (monoethanolammonium) is the counter-ion present in Alcohols C10-16, ethoxylated (1-2,5 EO) sulphated, monoeth salts (CAS 157627-92-4), MEA might have an impact on the toxicological properties of the registered substance. Therefore, the toxicological profile of MEA is considered. MEA is listed in Annex VI of Regulation 1272/2008. It is classified Acute Tox. 4; H302, H312 and H332 as well as Skin Corr. 1B; H314. Additionally a specific concentration limit is established for STOT SE3, H335 at concentrations ≥ 5% in Annex VI of Regulation 1272/2008. The effects of MEA on human health were assessed by the OECD in the SIDS initial assessment report (2013). No long-term toxicity study with MEA is available. However in a dietary two generation toxicity study with MEA the NOAEL was established at 300 mg/kg bw/d based on reduced body weight gain and food consumption at 1000 mg/kg bw/d. When this NOAEL is corrected for differences in molecular weight (61 g/mol for MEA and 343 g/mol for AES C12-14 MEA) this results in a NOAEL of 1686 mg/kg bw/d for AES C12-14 MEA. As the systemic NOAEL of AES (300 mg/kg bw/d) is lower than the corrected one of MEA, the systemic NOAEL established within the alkyl ether sulfates category is regarded to cover the repeated dose toxicity of MEA, too.

References:

Danish EPA - Environmental and Health Assessment of Substances in Household Detergents and Cosmetic Detergent Products (2001). Environmental Project No. 615, pp. 24-28

HERA (2003). Human & Environmental Risk Assessment on ingredients of European household cleaning products Alcohol Ethoxysulphates, Human Health Risk Assessment Draft, 2003. http: //www. heraproject. com.

SIDS initial assessment report, (2013); http://webnet.oecd.org/HPV/UI/SIDS_Details.aspx?key=8aefe41b-8499-4052-943f-f6dd6f8c5997&idx=0

Justification for classification or non-classification

The available data on repeated dose toxicity do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification.