Sulfuric acid, mono-C12-14 (even numbered)-alkyl esters, compds. with ethanolamine

Endpoint summary

Description of key information

Oral LD50 (OECD guideline 401), rat = 1800 mg/kg bw
Dermal LD50 (OECD guideline 402), rabbit > 2000 mg/kg bw (limit test) 
Acute toxicity by inhalation was not tested according to REGULATION (EC) No 1907/2006, Annex VIII, Section 8.5, Column 2.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP Guideline study According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Justification for type of information:

Refer to the Category Approach Justification document in point 13.

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

yes

Remarks:

lack on details of test substance within the study report. Missing details were however received by the sponsor.

GLP compliance:

yes

Test type:

fixed dose procedure

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

Doses:

500 and 2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 500 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All male and one female animal treated at 2000 mg/kg bw died in the course of the first day of the study.

Clinical signs:

other: At 2000 mg/kg bw: All animals showed hunched back, decrease in motor activity, ataxia and pallor. In addition the males also presented piloerection, and the females hypotonia. At 500 mg/kg bw: No clinical signs

Gross pathology:

One male of the 2000 mg/kg bw dose group showed hemoperitoneum, gastric and small intestine mucous membranes very congested, and the inside of this organs with bloody looking liquid.
One female of the 2000 mg/kg bw dose group showed dilation of the whole of the small and cecum intestines, without apparent lesions.

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 800 mg/kg bw

Quality of whole database:

The whole data base is conclusive and of high quality.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

Acute toxicity by inhalation was not tested according to REGULATION (EC) No 1907/2006, Annex VIII, Section 8.5, Column 2.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 June 2012 - 30 August 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

OECD Guideline study conducted in compliance with GLP regulations According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Details on strain: Wistar / Crl:WI (Han) SPF
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: approx. 9 weeks (males) and 13 weeks (females), respectively
- Weight at study initiation: mean (males): 273.0 ± 6.63g; mean (females): 225.8 ± 3.19g
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: single housing in Makrolon cage, type III
- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany
- Water (ad libitum): Tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22°C ± 3°C
- Humidity: 30 – 70%
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

deionized water

Details on dermal exposure:

TEST SITE
- Area of exposure: clipped epidermis (dorsal and dorsolateral parts of the trunk)
- % coverage: about 40 cm² (corresponds to at least 10% of the body surface)

REMOVAL OF TEST SUBSTANCE
- Washing: rinsing of the application site with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5.71 mL/kg bw
- Concentration (if solution): 35 g/100 mL
- Constant volume or concentration used: yes
- For solids, paste formed: no
- Form of application: solution in deionized water
- Test item preparation and homogenization: For better handling the test item was ground with mortar and pestle. The test item preparation was produced shortly before application by stirring with a magnetic stirrer.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 animals/sex/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals.

Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation.
The evaluation of skin reactions was performed according to Draize, J.H. (1959): Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas.

Body weight: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.

Pathology: Necropsy with gross-pathology examination on the last day of the observation period after sacrifice with CO2 in a chamber with increasing concentrations over time.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: No systemic clinical signs were observed during clinical examination.

Gross pathology:

There were no macroscopic pathological findings in the animals sacrificed at the end of observation period.

Other findings:

Local effects:
No local effects were observed.

Body weight changes:

Individual body weight changes
Dose [mg/kg bw]	2000
Sex	male
Animal No.	1	2	3	4	5	mean	sd.
Body weight at study day [g]
0	274	267	282	266	276	273.0	6.63
7	278	284	301	281	285	285.8	8.93
14	307	304	330	299	308	309.6	11.93

 

Individual body weight changes
Dose [mg/kg bw]	2000
Sex	female
Animal No.	1	2	3	4	5	mean	sd.
Body weight at study day [g]
0	224	222	230	225	228	225.8	3.19
7	231	220	234	226	230	228.2	5.40
14	237	238	240	239	238	238.4	1.14

sd. = standard deviation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The whole data base is conclusive and of high quality.

Additional information

There is only one acute oral toxicity study for C12-14AS MEA (CAS 90583-16-7) available. Therefore the endpoint acute toxicity is covered by read across to structurally related alkyl sulfates (AS), i.e. C12-14AS Na (CAS 85586-07-8), C10-16AS Na (CAS 68585-47-7), C12AS Na (CAS 151-21-3) and C12-13AS Na (CAS 91783-23-2) for acute oral toxicity and C10-16AS NH4 (CAS 68081-96-9), C10-16AS Mg (CAS 68081-97-0), C12-13AS K (CAS 91783-22-1) and C8AS Na (CAS 142-31-4) for the dermal route. The possibility of a read-across to other alkyl sulfates in accordance with Regulation (EC) No 1907/2006 Annex XI 1.5. Grouping of substances and read-across approach was assessed. In Annex XI 1.5 it is given that a read-across approach is possible for substances, whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity. The AS reported within the AS category show structural similarity. The most important common structural feature of the category members is the presence of a predominantly linear aliphatic hydrocarbon chain with a polar sulfate group, neutralized with a counterion. This structural feature confers the surfactant properties of the alkyl sulfates. The surfactant property of the members of the AS category in turn represent the predominant attribute in mediating effects on mammalian health. The AS of the AS category also have similar physicochemical, environmental and toxicological properties, validating the read-across approach within the category. The approach of grouping different AS for the evaluation of their effects on human health and the environment was also made by the OECD in the SIDS initial assessment profile [1] and by a voluntary industry program carrying out Human and Environmental Risk Assessments (HERA [2]), further supporting the read across approach between structurally related AS.

There is a substantial data base on ethanolamine (MEA) online available. MEA is listed in Annex VI of Regulation 1272/2008. It is classified as 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 [3]. MEA was found to have moderate acute oral toxicity but to have only low inhalation and dermal toxicity. LD50 values did not result in classification for acute dermal and inhalation exposure. As alkyl sulfates with a carbon chain length distribution C12-14 have to be classified as Acute Tox. 4; H302 the MEA content of C12-14AS MEA does not pose a further hazard for this endpoint. Additionally the concentration of freely available MEA in C12-14AS MEA (CAS 90583-16-7) is below 1%. Thus, unbound MEA will not contribute to the acute toxicity of AS. Therefore, no classification for these endpoints is required and read-across to alkyl sulfates with other counter ions is considered to be valid and reliable.

Acute oral toxicity 

There is one study for C12-14AS MEA (CAS 90583-16-7) and four relevant studies for the read-across substances C12-14AS Na (CAS 85586-07-8), C10-16AS Na (CAS 68585-47-7), C12AS Na (CAS 151-21-3) and C12-13AS Na (CAS 91783-23-2) addressing acute oral toxicity available.

The study with C12-14AS MEA (CAS 90583-16-7) was performed similar to OECD Guideline 401. Groups of 10 male Wistar rats were administered different doses of the test item and observed for 8 days. Only limited information on test conditions and experimental methods and results are available. The LD50 based on the test material was 5620 mg/kg bw. Based on active substance the LD50 was 1686 mg/kg bw.

The key study conducted with C12-14AS Na (CAS 85586-07-8), was performed similar to OECD Guideline 420 on Wistar rats (Puig, 1994). Five Wistar rats of both sexes were dosed with the test substance at 500 and 2000 mg/kg bw via gavage. All males and one female of the top dose group died within 24 h after the application of the test substance. No clinical signs of toxicity were seen in animals of the low dose group. Clinical signs of toxicity at 2000 mg/kg comprised hunched back, decrease in motor activity, ataxia and pallor. In addition the males had piloerection and the females hypotonia. Upon necropsy haemoperitoneum, congested mucous membranes of the gastro-intestinal tract, and bloody looking liquid inside these organs occurred in one of the males which died during the course of the study. One female of the 2000 mg/kg bw dose group showed dilation of the small intestines and caecum. No further findings were observed upon necropsy. The LD50 was greater than 500 but below 2000 mg/kg bw.

A study conducted with C10-16AS Na (CAS 68585-47-7) was performed similar to OECD Guideline 401 (Cawley, 1978). Five Spraque-Dawley rats of each sex were dosed with the test substance at the limit dose of 2000 mg/kg bw via gavage and observed for mortality and clinical signs for 14 days. One female died after application of the test substance. No clinical signs of toxicity were seen in any animal. The LD50 was determined to be greater than 2000 mg/kg bw.

A further study was conducted with C12AS Na (CAS 151-21-3, analytical purity 98%) similar to OECD Guideline 401 with acceptable restrictions on Wistar rats (Potokar, 1983). Both sexes were dosed with the test substance via gavage. Mortalities occurred but no details were available. Clinical signs of toxicity comprised diarrhoea, spastic gait, decreased activity, lateral position, hunched posture, laboured respiration and coma. Necropsy of dead animals revealed haemorrhages in gastro-intestinal tract and vascular congestion in the liver. No findings were observed in animals sacrificed upon study termination. The LD50 was given as 1200 mg/kg bw for males and females, 977 mg/kg bw for females and 1427 mg/kg for males. 

A further study performed similar to OECD Guideline 401 was conducted with C12-13AS Na (CAS 91783-23-2, analytical purity 26.8%). Five Wistar rats of each sex were dosed with the test substance at 880, 1040, 1260 and 1500 mg/kg bw via gavage and observed for mortalities for 14 days (Sasol, 1984). At the lowest dose no mortalities occurred. In the 1040, 1260 and 1500 mg/kg bw dose groups 0/5, 3/5 and 5/5 males and 2/5, 5/5 and 5/5 females died within 14 days. No data on clinical signs of toxicity are available. The LD50 was determined to be 1230 mg/kg bw for males and 1063 mg/kg bw for females.

In the following a fixed dose for the LD50-value is discussed. Looking at the acute oral toxicity of AS in rodents it can be seen that the toxicity decreases with increasing chain length (see Category Justification Report & HERA report). Evaluations of test materials comprising mixtures of chain lengths also showed a trend to lower toxicity as the proportion of longer chain lengths in the test material increases.

Using the OECD 420 fixed dose method no deaths at 500 mg/kg bw and 60% deaths at 2000 mg/kg bw occurred, resulting in a LD50 presumably greater than 1000 mg/kg bw for C12-14AS Na (Puig, 1994). The LD50 for C12AS in a study with 98% active ingredient was 1200 mg/kg bw (Potokar, 1983) for males and females. The LD50 for C12-13AS was in a comparable dose range (Sasol, 1984). Both substances are more toxic than C12-14AS Na and hence represent a worst-case. Moreover, taking into account that the study with 85% active ingredient of C10-16AS resulted in a LD50 of greater than 2000 mg/kg bw (Cawley, 1978), the adjusted LD50 for C12-14AS (100%) would be 1800 mg/kg bw. This value is comparable to the value calculated for neat C12-14AS MEA in as study with lower reliability (Kästner, 1971).

Hence, a LD50 of 1800 mg/kg bw is also justified for C12-14AS MEA (CAS 90583-16-7).

Acute dermal toxicity

Regarding the acute dermal toxicity four studies are available for the read-across substances C10-16AS NH4 (CAS 68081-96-9), C10-16AS Mg (CAS 68081-97-0), C12-13AS K (CAS 91783-22-1) and C8AS Na (CAS 142-31-4).

The key study was conducted with C8AS Na (CAS 142-31-4) according to OECD Guideline 402 (BASF, 2012). 5 Wistar rats per sex were trated with 2000 mg/kg bw under semi occlusive conditions for 24 h. No mortality occurred, no sign of systemic toxicity and of local irritation was observed.

The study conducted with C10-16AS NH4 (CAS 68081-96-9) was performed as limit test conducted similar to OECD Guideline 402 with 3 male and 3 female New Zealand White rabbits (Frank, 1975). The test substance (analytical purity 25.1%) was applied at 2000 mg/kg bw for 24 h under occlusive conditions. No mortalities occurred. Clinical signs of toxicity comprised severe erythema and slight eschar formation at 24 h, necrosis on day 2–14 with sloughing of the skin on day 8–14 and hyper-pigmentation of new skin by day 14. No signs of systemic toxicity were observed. Hence, the LD50 value is greater than 2000 mg/kg bw based on the test material and greater than 500 mg/kg bw based on active ingredient.

The study conducted with C10-16AS Mg (CAS 68081-97-0) was performed as limit test conducted similar to OECD Guideline 402 with 3 male and 3 female New Zealand White rabbits (Frank, 1975). The test substance (analytical purity 23.5%) was applied at 2000 mg/kg bw for 24 h under occlusive conditions. No mortalities occurred. Clinical signs of toxicity comprised severe erythema and eschar formation at 24 h, necrosis by Day 5–21 and necrotic tissues sloughed and leaving skin hyper-pigmented at Day 21. Hence, the LD50 value is greater than 2000 mg/kg bw based on the test material and greater 500 mg/kg bw based on the active ingredient.

Another study similar to OECD Guideline 402 was performed with C12-13AS K (CAS 91783-22-1) on three male and three female New Zealand White rabbits (Benedict, 1978). Both sexes were dosed at 2000 mg/kg bw (analytical purity 25%) under occlusive conditions for 24 h. No mortalities occurred during the conduct of the study. Findings within this study comprised moderate to severe erythema, edema, and atonia, desquamation and fissuring by day 6 and eschar formation and exfoliation. Based on the above mentioned findings the LD50 was greater than 2000 mg/kg bw based on test material and greater 500 mg/kg bw based on active ingredient.

Within the studies mentioned above no mortalities or signs of systemic toxicity occurred after dermal application of alkyl sulfates with varying carbon chain length. Taken into account the generally low toxicity after oral application and the low dermal absorption rate of alkyl sulfates (approx. 1%) this result was expected. In agreement, no classification for acute dermal toxicity is required. For details on absorption please see section Toxicokinetics, metabolim and distribution.

Acute inhalation toxicity

No studies for acute inhalation toxicity are available. However, testing the potential of acute toxicity via inhalation route of C12-14AS MEA (CAS 90583-16-7) is considered to be not justified. According to Regulation (EC) No 1907/2006, Annex VIII, Section 8.5, Column 2, in addition to the oral route (8.5.1), for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. As information under 8.5.3 (dermal route) is provided, the requirement is fulfilled by using the most appropriate route of exposure.

AS is mainly used in liquid media. Due to its very low vapour pressure [2], inhalation is not viewed as a significant route of exposure. Inhalation of AS may occur via aerosols generated by spray cleaners or by detergent dusts (e.g. washing powder). Taking into account that the acute toxicity of AS is generally low but that the substance is irritating to skin and eyes at concentrations of 10% or higher, no additional information is expected from testing for acute inhalation toxicity as the predominant effect will be local irritation rather than systemic toxicity. However, as the neat substance has to be classified as harmful if swallowed the substance will also be classified as harmful if inhaled (H332: Acute tox 4 and R20, respectively) in case the substance is available as neat powder.

 

[1] SIDS initial assessment profile, (2007); http://www.aciscience.org/docs/Alkyl_Sulfates_Final_SIAP.pdf

[2] (HERA Draft report, 2002); http://www.heraproject.com/files/3-HH-04-%20HERA%20AS%20HH%20web%20wd.pdf

[3] SIDS initial assessment report, (2013);

http://webnet.oecd.org/HPV/UI/SIDS_Details.aspx?key=8aefe41b-8499-4052-943f-f6dd6f8c5997&idx=0

Justification for selection of acute toxicity – oral endpoint
Reliable OECD guideline study.

Justification for selection of acute toxicity – dermal endpoint
Reliable OECD guideline study.

Justification for classification or non-classification

The available data on acute oral toxicity meet the criteria for classification as Acute Toxicity Category 4 (H302) according to Regulation (EC) 1272/2008 and as Xn (R22) according to Directive 67/548/EEC. The available data on acute dermal toxicity do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.

Exposure to solid C12-14AS MEA (CAS 90583-16-7) with a bulk density < 400g/L may occur. As worst case assumption this material is classified as Acute Toxicity Category 4 (H332) according to Regulation (EC) 1272/2008 and as Xn (R20) according to Directive 67/548/EEC.