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Administrative data

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 data for subchronic toxicity were available from an oral (diet) 90-day toxicity study in rats with read-

across substance 'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered),

C18unsaturated alkyl) tetrasodium salts' dosed at 0.5, 2 and 8 (reduced to 4) g/kg bw.day. The study

showed decreased body weight gain, feed consumption and food efficiency at the mid dose and high

dose levels and increased SGOT and SGPT (liver enzymes) at the high dose. Further hematuria was

seen in the mid and high dose rats, various organ weights were decreased (e.g. decrease in adrenal

and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group)

and lower urinary tract pathology was seen in 2 high dosed rats. It thus appeared that the NOEL in the

rat was below 0.50 g/kg/day, however 0.5g or 500 mg act.ingr./kg bw/day can be considered as NOAEL.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

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:
reference to same study
Reason / purpose:
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
Critical effects observed:
not specified

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:
sub-chronic toxicity: oral
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:
read-across source
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.
Mortality:
mortality observed, treatment-related
Description (incidence):
Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
[Trade name] depressed significantly the rate of body weight gain in the mid dose and high dose rats under the conditions of this experiment. This depression appeared more pronounced in the male than in the female rats.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
During weeks four through nine the feed consumption of the high dose male animals was significantly depressed. The other groups showed only an occasional significant depression.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
The feed efficiency of the mid dose and high dose test groups is decidedly different from that of the control group, more so in the male than in the female rats.
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):
Some rats in all levels exhibited slightly elevated SGOT values and 4 high dose rats exhibited elevated SGPT levels. Histologically, however, there was no basis for these elevations.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Some of the rats in the mid dose and high dose group displayed hematuria to a greater or lesser degree. Their urinary system was not remarkably changed histologically. Only one rat, #5103, high dose group, displayed hematuria and moderate acute cystitis.
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Decrease in thyroid weight in males of high dose group; decrease in adrenal and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group; decrease in brain weight in males of mid dose and high dose group.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat had hydrophilic degeneration of liver and kidneys, 1 high dose rat had hepatic lipidosis.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat had hydrophilic degeneration of liver and kidneys, 1 high dose rat had hepatic lipidosis.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Due to decrease in body weight and feed consumption in the high dose group (8.00 g/kg/day) the level was lowered to 4.00 g/kg/day for weeks five through the termination of the study. Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.

BODY WEIGHT AND WEIGHT GAIN
[Trade name] depressed significantly the rate of body weight gain in the mid dose and high dose rats under the conditions of this experiment. This depression appeared more pronounced in the male than in the female rats.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
During weeks four through nine the feed consumption of the high dose male animals was significantly depressed. The other groups showed only an occasional significant depression.

FOOD EFFICIENCY
The feed efficiency of the mid dose and high dose test groups was decidedly different from that of the control group, more so in the male than in the female rats.

HAEMATOLOGY
All rats had normal hematocrit concentrations for the duration of the experiment
All rats had normal hemoglobin concentrations for the duration of the experiment
Rat #5097 (4.00 g/kg/day group) had a WBC of 35000 cells/mL at six weeks but by three months its WBC was normal. The rest of the rats had normal white cell counts at all times examined.
All rats examined at six and 13 weeks displayed lymphocytosis.
The RBC’s of all animals were morphologically normal at all times.

CLINICAL CHEMISTRY
All rats had normal blood glucose values when examined.
All rats examined had normal BUN values.
Some rats in all levels exhibited slightly elevated SGOT values. Histologically, however, there was no basis for these elevations.
Four high dose rats exhibited elevated SGPT levels. Histologically, however, there was no basis for these elevations.
All rats examined had normal GGPT values.

URINALYSIS
Some of the rats in the mid dose and high dose group displayed hematuria to a greater or lesser degree. Their urinary system, however, was not remarkable histologically. Only one rat, #5103, high dose group, displayed hematuria and moderate acute cystitis

ORGAN WEIGHTS
Cfr. Table 3 and 4 (absolute & relative organ weights)

GROSS PATHOLOGY
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat hydrophilic degeneration of liver and kidneys, 1 high dose rat hepatic lipidosis.
Key result
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
act. ingr.
Remarks:
read-across test item
Sex:
male/female
Basis for effect level:
other: overall effects
Critical effects observed:
not specified

Table 1. Body weight data for rats receiving [Trade name] for 13 weeks

Dose levels (g/kg).

Mean male Body Weight (g)

Weight Gain (g)

Mean female Body Weight (g)

Weight Gain (g)

Initial

Terminal

Initial

Terminal

0

68.4

509.0

440.6

60.0

289.3

229.3

0.50

68.3

505.6

437.3

60.1

276.7

216.6

2.00

68.4

434.5*

366.1

60.1

249.9*

189.8

8.00 (4.00)

68.2

269.8*

192.6

59.8

189.1*

129.3

* Statistically significant from control at P=0.05.

Table 2. Feed consumption for rats receiving [Trade name] for 13 weeks (g/rat/week)

Week

on

diet

Males

Females

Dose levels (g/kg)

Dose levels (g/kg)

0

0.50

2.00

8.00 (4.00)

0

0.50

2.00

8.00 (4.00)

1

103.8

97.2

112.5

101.5

83.3

90.3

86.8

84.0

2

124.8

119.0

133.5

137.0

103.5

116.5

106.3

115.3

3

157.3

157.8

162.8

163.0

134.8

144.3

150.5*

156.5*

4

157.3

135.0*

132.5*

186.1*

137.5

123.8

125.5

122.3

5

199.0

189.0

195.3

152.1*

158.3

138.0*

140.0*

150.8

6

185.7

185.7

190.1

162.8

145.0

159.2

168.8*

138.0

7

184.8

183.0

188.0

206.6*

130.8

131.0

137.1

170.8*

8

185.0

186.0

183.0

196.1*

150.3

138.5

152.8

164.3

9

175.3

178.5

184.3

194.2*

149.5

139.8

136.5*

166.0

10

181.0

179.5

184.0

176.3

134.8

129.3

138.8

150.8

11

182.5

191.0

183.7

177.1

147.5

153.2

143.2

164.5

12

179.0

178.4

176.3

182.4

160.0

145.0

158.3

172.8

13

190.0

188.9

177.0

160.5*

142.5

126.8

131.0

167.1

Mean

169.7

166.8

169.5

168.9

136.8

133.5

136.6

147.9

* Statistically significant from control at P=0.05.

Table 3. Summary of Mean Body Weights and Organ Weights on rats receiving [Trade name] in their diet for 13 weeks

Sex

Dose

Level.

g/kg

Terminal

Body Weight

   (g)

Organ Weights

Thyroids

Heart

Liver

Adrenals

Kidneys

Gonads

Pituitary

Brain

Male

0

503.9

0.034

1.262

20.390

0.060

3.527

3.361

0.013

2.216

 

0.50

503.7

0.031

1.274

22.540

0.060

3.566

3.491

0.013

2.107

 

2.00

443.3*

0.033

1.158

15.884*

0.064

2.984*

3.179

0.014

2.084*

 

8.00(4)

286.9*

0.025*

0.809*

9.890*

0.046*

1.926*

2.709*

0.010

1.906*

 

Female

0

301.6

0.033

0.879

11.326

0.085

1.966

0.152

0.017

1.969

 

0.50

280.9

0.034

0.799

9.759*

0.080

1.900

0.169

0.017

1.969

 

2.00

254.3*

0.031

0.755*

10.529

0.077

1.912

0.153

0.017

1.897

 

8.00(4)

200.2*

0.032

0.636*

8.316*

0.061*

1.582*

0.101*

0.012*

1.938

*Statistically significant from control at P=0.05

Table 4. Organ-to-body weight ratios for male and female beagle dogs receiving [Trade name] in their diet for 13 week

 

Dose

Levels

g/kg

Terminal

Body Weight

   (g)

(Grams of organ weight/kilograms of body weight)

 

Sex

Thyroids

Heart

Liver

Adrenals

Kidneys

Gonads

Pituitary

Brain

Male

0

503.9

0.067

2.504

40.464

0.119

6.999

6.670

0.026

4.398

 

0.50

503.7

0.073

2.529

44.749

0.119

7.080

6.931

0.026

4.183

 

2.00

443.3*

0.074

2.612

35.831

0.144

6.731

7.171

0.032

4.701

 

8.00(4)

286.9*

0.087

2.820

34.472

0.160

6.713

9.442

0.035

6.643

 

Female

0

301.6

0.109

2.914

37.553

0.282

6.519

0.504

0.056

6.529

 

0.50

280.9

0.121

2.844

34.742

0.285

6.764

0.602

0.061

7.010

 

2.00

  254.3*

0.122

2.969

41.404

0.303

7.519

0.602

0.067

7.460

 

8.00(4)

 200.2*

0.160

3.177

41.538

0.305

7.902

0.504

0.060

9.680

*Statistically significant from control at P=0.05.

 

 

Conclusions:
These studies have shown that the read-across test item, when fed to random bred albino rats under the conditions of this experiment, decreased the rate of body weight gain, feed consumption and food efficiency at the mid dose and high dose levels, and increased SGOT and SGPT at the high doses. Further hematuria was seen in mid and high dosed rats, various organ weights were decreased and lower urinary tract pathology was seen in 2 high dosed rats.It thus appeared that the no effect level in the rat is below 0.50 g/kg/day, however 0.50g act.ingr./kg bw/day can be considered as NOAEL.
Executive summary:

Sprague-Dawley albino rats, 80 of each sex, were fed a control diet, or 0.50, 2.00 or 8.00 g/kg /day of read-across test item, containing 35.8% active ingredient, mixed in the diet. The high dose was reduced to 4.00 g/kg/day of test item mixed in the diet for weeks five through to termination. The rats were carefully observed for the duration of the experiment and pertinent hematological and biochemical parameters were conducted while the experiment was in progress. At the conclusion of the experiment all rats were necropsied and all organs and tissues were examined histologically from ten rats (five male and five female) from the control and high dose groups. These studies have shown that the read-across test item, when fed to random bred albino rats under the conditions of this experiment, decreased body weight gain, feed consumption and food efficiency at the mid dose and high dose levels and increased SGOT and SGPT at the high doses. Further hematuria was seen in the mid and high dose rats, various organ weights were decreased (e.g. decrease in adrenal and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group) and lower urinary tract pathology was seen in 2 high dosed rats. It thus appeared that the no effect level in the rat is below 0.50 g/kg/day, however 0.5g act.ingr./kg bw/day can be considered as NOAEL

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 quality (Klimisch 2)

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

No test data were available for current substance, however read across data were available from N2 and N3 subgroup members. For these substances, comparative 14 -day dose range finding studies were available. Justification for read across within the subgroups of N-containing sulphosuccinates (N2 and N3) 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'.

 

Subchronic toxicity

Data were available for read across substance 'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium salts'

- A supporting 14-day dose-range-finding was conducted with test item containing 34.4% active ingredient (Hansen, 2013d). Male and female rats were treated orally with 100, 300 or 1000 mg act. ing. /kg bw/day. No rats died prematurely nor revealed any test item-related changes in behaviour, external appearance or faeces. No changes in body weight and body weight gain, food and drinking water consumption or for relative and absolute organ weights were noted at any of the tested dose levels. Macroscopic examination revealed no test item-related changes at any of the tested dose levels. Maximum tolerated dose was 1000 mg/kg bw.

- In a key subchronic repeated dose toxicity study, 160 Sprague-Dawley albino rats, 80 of each sex, were fed a control diet, or 0.50, 2.00 or 8.00 g/kg /day of test item mixed in the diet (Tegeris and Underwood, 1976a). The liquid test item contained 35.8% active ingredient, which was taken into account for the dosages. The high dose was reduced to 4.00 g/kg/day mixed in the diet for weeks five through to termination. The study showed decreased body weight gain, feed consumption and food efficiency at the mid dose and high dose levels and increased SGOT and SGPT at the high dose. Further hematuria was seen in the mid and high dose rats, various organ weights were decreased (e.g. decrease in adrenal and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group) and lower urinary tract pathology was seen in 2 high dosed rats. It thus appeared that the NOEL in the rat was below 0.50 g/kg/day, however 0.5g act.ingr./kg bw/day can be considered as NOAEL.

- In another subchronic repeated dose toxicity study, test item containing 35.8% active ingredient was given in the diet to purebred beagle dogs for ninety days (Tegeris and Underwood, 1976b). Thirty-two purebred beagle dogs, sixteen of each sex, with an average age of three to four months, were fed the control diet or 0.062, 0.250 or 1.000 g act.ingr. /kg bw/day thoroughly mixed in the diet. The dogs were carefully observed for the duration of the experiment and several hematological and biochemical parameters were conducted while the experiment was in progress. At the conclusion of the experiment all dogs were necropsied and all organs and tissues were examined histologically. These studies have shown that the test substance interfered with the average daily feed consumption of the mid-dose female and high dose test dogs and decreased the rate of bodyweight gain of the high dose test dogs when fed to them under the conditions of this experiment. Otherwise it was harmless up to 1 g/kg bw/day; a NOAEL of 0.250 g/kg bw/day can be considered.

- In conclusion, NOAEL-level of 500 mg/kg bw for systemic toxicity was observed in a dietary subchronic toxicity study in the rat with read-across substance 'Aspartic acid, N-(3-carboxy-1-oxo-sulfopropyl)-N-(C16-C18 (even numbered), C18unsaturated alkyl) tetrasodium 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 the most reliable and conservative 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 selection of repeated dose toxicity via oral route - systemic effects endpoint: Key study 

Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: liver; digestive: stomach

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.