Butanedioic acid, sulfo-, C-[2-[(1-oxododecyl)amino]ethyl] ester, monosodium salt

Endpoint summary

Description of key information

Key data for subacute toxicity were available from an oral (gavage) OECD 422 study in rats with read-across substance 'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts', at dose levels given by oral gavage of 100, 300 and 1000 mg active ingredient/kg bw/day. No relevant effects were observed at 100 and 300 mg/kg bw. At the dose of 1000 mg/kg bw, decreased body weight, increased serum ALAT and decreased

serum albumin were observed as systemic changes, whereas macroscopic and microscopic stomach changes were observed as local changes, the latter without relevance to humans. NOAEL for paternal/ maternal toxicity was 300 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

see attached read-across justification

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

read-across source

Frequency of treatment:

daily

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

salivation in one male rat at 1000 mg/kg bw/day

Mortality:

mortality observed, treatment-related

Description (incidence):

salivation in one male rat at 1000 mg/kg bw/day

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

slight reduction in male and female rats at 1000 mg/kg bw/day

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

slight increase in food consumption in male rats dosed at 1000 mg/kg bw/day

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

increased ALAT in male and female rats dosed at 1000 mg/kg bw/day; decrease in albumin in male rats dosed at 1000 mg/kg bw/day;

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

changes in the stomach from 2 male animals dosed at 1000 mg/kg bw/day: whitish thickening (cardia), yellowish contents

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

squamous cell hyperplasia and in the non glandular mucosa and acute inflammation in male and female rats dosed at 1000 mg/kg bw/day; pulmonary congestion in male and female rats dosed at 1000 mg/kg bw/day

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No test item-related premature death was noted in any treatment group (100, 300 and 1000 mg/kg bw/day).
No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day).
A slightly increased salivation was noted in one male rat, no further signs of clinical toxicity were noted for the male and female rats of the high dose group (1000 mg/kg bw/day).
No test item related influence was noted for the male and female rats of all treatment groups (100, 300 and 1000 mg/kg bw/day) during the observational and functional (grip strength and spontaneous motility) neurological screenings.

BODY WEIGHT AND WEIGHT GAIN
A slight reduction in body weight was noted for the male and female rats of the high dose group (1000 mg/kg bw/day). For the male rats the reduction in body weight was noted from test day 8 (4.4%) until test day 36 (5.5%) and for the female rats from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
A slightly statistically significant (p ≤ 0.01) increase in relative food consumption by 10.3% was noted in the high dose males during the 2nd test week. This was caused by the reduced body weight of the rats of the high dose group.
No influence on food consumption was noted in any treatment group in the females.

HAEMATOLOGY
No test item-related influence was noted.

CLINICAL CHEMISTRY
The laboratory examinations revealed an increased ALAT activity for the male and female rats of the high dose group (1000 mg/kg bw/day) and a decrease in the albumin concentration for the male rats of the high dose group.

NEUROBEHAVIOUR
No test item-related influence was noted .

ORGAN WEIGHTS
No test item-related influence was noted .

GROSS PATHOLOGY
No test item-related changes were noted during the macroscopic inspection at autopsy with the exception of changes in the stomach from 2 male animals from the high dose group (1000 mg/kg bw/day) ), which were considered to be test item-related.
No test item related findings were noted in the female animals.

HISTOPATHOLOGY: NON-NEOPLASTIC (restricted to dose groups 1 and 4)
A statistically significant (p≤0.01) occurence of squamous cell hyperplasia in the non glandular mucosa of the forestomach was noted for the male and female rats (5 of 5 each) of the high dose group (1000 mg/kg bw/day). Occasionally the squamous cell hyperplasia with subsequent hyperkeratinization was associated with acute inflammation of the submucosa in the non-glandular stomach (for 2/5 males and 1/5 females).
A pulmonary congestion was found in 4 of 5 male animals, which was statistically significant (p≤0.05) in comparison to the control group (0/5).
No microscopic changes were noted for the reproductive organs of the male and female rats of the high dose group (1000 mg/kg bw/day).

Evaluation of reproduction parameters: see section 7.8.1 & 7.8.2

Key result

Dose descriptor:

NOAEL

Remarks:

Parental generation F0

Effect level:

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

Based on:

act. ingr.

Remarks:

read-across test item

Sex:

male/female

Basis for effect level:

other: Systemic effects

Key result

Critical effects observed:

no

Table 1. Mean body weight males

Body Weight (g)
Sex: Male	Day(s) Relative to Start Date
	1	8v	15	22v	29v	36v
Group 1: control	293.02	334.59	350.28	386.70	410.83	440.86
Group 2: 100 mg/kg	293.46	328.96	347.75	380.93	405.63	436.65
Group 3: 300 mg/kg	294.16	336.32	353.72	392.53	416.39	446.90
Group 4: 1000 mg/kg	293.14	319.93*	338.13	367.75	389.07	416.42

Statistical Test Dunnett’s Test (Anova)

Group Factor Dunnett’s Test (Anova): v –Statistical Test: Analysis of Variance p<0.05

*-Statistical Test: Dunnett 2 Sided p<0.05

 

Table 2. Mean body weight females

Body Weight (g)
Sex: Female	Day(s) Relative to Start Date			Day(s) Relative to Mating (L)				Day(s) Relative to Littering (A)
	1	8	15	0	7v	14v	20	1vv	4v
Group 1: control	181.25	200.61	207.74	231.36	272.16	301.56	365.20	283.99	301.20
Group 2: 100 mg/kg	180.93	197.80	208.12	226.64	269.64	303.46	376.18	290.81	302.18
Group 3: 300 mg/kg	180.90	194.82	204.24	224.81	254.78	289.41	356.96	271.43	284.22
Group 4: 1000 mg/kg	181.33	193.29	200.78	213.67	245.74*	277.59*	341.34	258.20*	272.58*

Statistical Test Dunnett’s Test (Anova)

Group Factor Dunnett’s Test (Anova): v –Statistical Test: Analysis of Variance p<0.05

                                                                vv- Statistical Test: Analysis of Variance p<0.01

*-Statistical Test: Dunnett 2 Sided p<0.05

 

Table 3. Mean Biochemical Parameters Males and Females (Albumin and ALAT)

Biochemical Parameters
Sex: Male	Albumin (g/L)	ALAT (U/L)v	Sex: Female	Albumin (g/L)	ALAT (U/L)vv
Group 1: control	31.98	38.0	Group 1: control	33.14	38.4
Group 2: 100 mg/kg	31.64	38.6	Group 2: 100 mg/kg	32.80	36.4
Group 3: 300 mg/kg	31.18	43.4	Group 3: 300 mg/kg	33.74	34.6
Group 4: 1000 mg/kg	30.60**	63.2**	Group 4: 1000 mg/kg	32.18	56.2**

Statistical Test Dunnett’s Test (Anova)

Group Factor Dunnett’s Test (Anova): v –Statistical Test: Analysis of Variance p<0.05

                                                                    vv- Statistical Test: Analysis of Variance p<0.01

*-Statistical Test: Dunnett 2 Sided p<0.05

**- Statistical Test: Dunnett 2 Sided p<0.01

Table 4.Histopathology Males and Females

Sex	Male				Female
Group	Gr.1	Gr.2	Gr.3	Gr.4	Gr.1	Gr.2	Gr.3	Gr.4
Number of Animals	5			5	5			5
Number of Completed Animals	5			5	5			5
Lungs
congestion	0			4*	2			1
Stomach
No abnormalities detected	5			0	5			0
Non-glandular; submucosa; acute inflammation -slight -moderate	0 0			1 1	0 0			0 1
Non-glandular; squamous cell hyperplasia -slight -moderate -marked	0 0 0 0			5** 1 3 1	0 0 0 0			5** 2 2 1
Non-glandular; keratopurulent debris	0			2	0	0

Fisher’s Two-Tailed Exact Test Performed:

*= 5% Significance

**= 1% Significance

Conclusions:

NOAEL (no-observed-adverse-effect level) of the parental generation: 300 mg/kg bw/day, p.o.

Executive summary:

The aim of the study was to obtain information on possible effects of the read-across test item on general toxicity, reproduction and/or development according to OECD guideline 422. The read-across test item was administered orally by gavage to rats at dose levels of 100, 300 and 1000 mg active ingredient/kg bw/day. The application started two weeks before mating on test day one and ended on the day or one day before sacrifice. Day of sacrifice was on test day 37 for the male rats and between lactation day 4 and 7 for the female rats. No test item-related premature death was noted in any treatment group (100, 300 and 1000 mg/kg bw/day).

No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day). A slightly increased salivation was noted in one male rat, no further signs of clinical toxicity were noted for the male and female rats of the high dose group (1000 mg/kg bw/day). No test item related influence was noted for the male and female rats of all treatment groups (100, 300 and 1000 mg/kg bw/day) during the observational and functional (grip strength and spontaneous motility) neurological screenings. A slight reduction in body weight was noted for the male and female rats of the high dose group (1000 mg/kg bw/day). For the male rats the reduction in body weight was noted from test day 8 (4.4%) until test day 36 (5.5%) and for the female rats from gestation day 0 (7.6%) until lactation day 4 (9.5%). The body weight at autopsy was reduced accordingly.

The laboratory examinations revealed an increased ALAT activity for the male and female rats of the high dose group (1000 mg/kg bw/day) and a decrease in the albumin concentration for the male rats of the high dose group.

No test item-related changes were noted during the macroscopic inspection at autopsy with the exception of changes in the stomach from 2 male animals from the high dose group (1000 mg/kg bw/day). Microscopic examination revealed the occurrence of squamous cell hyperplasia in the non-glandular mucosa and acute inflammation of the forestomach from the male and female rats of the high dose group (1000 mg test item/kg bw/day). Further microscopic findings occurred in form of pulmonary congestion in the male rats from the high dose group. NOAEL (no-observed-adverse-effect level) for repeated dose toxicity: 300 mg/kg bw/day, p.o.

Effects on reproduction parameters and organs (see section 7.8.1).

Effects on the development of the F1offsprings (pups) (see section 7.8.2).

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

300 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Reliable without restriction

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Mode of Action Analysis / Human Relevance Framework

In the absence of any evidence for species specific effects or modes of action (beside changes in the stomach) the effects observed in animals and the absence of effects are regarded as relevant for humans.

Additional information

No test data were available for current substance, however read across data were available from N2 subgroup members. For these substance, comparative 14 -day dose range finding studa was available. Justification for read across within the subgroups of N-containing sulphosuccinates (N2) is documented in a separate document attached in Section 13.

 

Subacute toxicity

Data were available for read across substance 'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts'.

- A key study for repeated dose toxicity was performed by means of an oral combined repeated dose and reproduction/development screening study according to OECD guideline 422 (Hansen, 2013c). The test item was administered orally by gavage to rats with a liquid formulation containing 41.5% active ingredient at dose levels of 100, 300 and 1000 mg/kg bw/day for at least 28 days in male rats and at least 39 days in females. No test item-related premature death was noted in any treatment group. No signs of clinical toxicity were noted for the male and female rats of the low and intermediate dose groups (100 and 300 mg/kg bw/day), whereas slightly increased salivation was noted in one male rat at 1000 mg/kg bw/day. No observational and functional neurological findings were seen up to the highest dose group. A slight reduction in body weight was noted for the male and female rats dosed at 1000 mg/kg bw/day. The laboratory examinations revealed an increased serum ALAT activity for the male and female rats dosed at 1000 mg/kg bw/day, and a decreased serum albumin concentration for the male rats of the high dose group. No test item-related changes were noted during the macroscopic inspection at autopsy with the exception of changes in the stomach from 2 male animals from the high dose group. Microscopic examination revealed the occurrence of squamous cell hyperplasia in the non-glandular mucosa of the forestomach and acute inflammation in the male and female rats of the high dose group. These changes are considered to be local, and not relevant for humans as humans lack a forestomach. NOAEL for paternal/maternal toxicity was 300 mg/kg bw/day.

- In a supporting 14-day dose-range-finding study the dose levels were selected for a combined repeated dose and reproduction/developmental toxicity screening test (Hansen, 2013c). 5 Male and 5 female rats were treated once daily with a liquid formulation containing 41.5% active ingredient at dose levels of 100, 300 and 1000 mg act.ingr./kg bw/day by oral gavage administration. None of the animals died prematurely. Salivation was noted for 2 of 5 male animals treated with 1000 mg/kg bw/day starting on day 9 and increased faeces was noted for 3 of 5 male and 2 of 5 female high dosed animals starting on test day 5. The food consumption of the male and female animals treated with 1000 mg/kg bw/day was slightly increased by 9% for the males and by 10% for the females in test week 2. None of the male and female rats treated orally with 100, 300 or 1000 mg/kg bw/day revealed any test item-related changes in body weight, body weight gain as well as relative and absolute organ weights or at macroscopic inspection at necropsy. NOAEL was 300 mg/kg bw/day.

- In conclusion, NOAEL-level of 300 mg/kg bw for systemic toxicity was observed in an oral gavage combined repeated dose and reproductive/developmental screening study in the rat which was performed with read-across substance 'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts'.

 

 

Conclusion

- The NOAEL of 300 mg/kg bw in the OECD 422 study with 'Butanedioic acid, 2(or 3)-sulfo-, 4-[2-[(1-oxo(C12-C18(even numbered) and C18unsaturated)alkyl))amino]ethyl]esters, disodium salts' was considered as reliable and relevant value, therefore this was selected as the descriptor for DNEL calculations.

- Further information supporting the safety of the test substance is provided in the read across justification for the N2 and N3 subgroups, showing that all substances in these groups had similar NOAELs (justification with data matrix separately attached in Section 13).

 

Justification for classification or non-classification

Based on these results and according to CLP (No. 1272/2008 of 16 December 2008), the test substance does not have to be classified and has no obligatory labelling requirement for repeated dose toxicity.