Amines, C12-18-alkyl, reaction products with acrylic acid, sodium salts

Endpoint summary

Description of key information

Oral:
Subacute toxicity study, 28days, oral, rat (OECD 422), up to 600mg/kg bw/day:  NOAEL =  160 mg/kg bw/day
Dermal and inhalation:
No reliable data available

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

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Apr - 28 Nov 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD test guideline and GLP compliant study, performed on a test material with the same alkyl chain distribution.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted 22. March 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650, July 2000

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

The study was performed on a commercial product (aqueous solution) as the test item is manufactured and used in a liquid form.

Species:

rat

Strain:

other: HanRcc: WIST(SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories Ltd., Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: mean: males 294 - 344 g; females 176 - 221 g
- Housing: individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libitum
- Water: community tap-water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

highly purified

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The dosage formulations were prepared weekly prior to administration using the test item as supplied by the Sponsor and using a factor of 3.72 taking in account the purity of 87% and the content of the active ingredient of 30.89% (CAS 3655-00-3). Sodium coco β-iminodipropionate (CAS 3655-00-3) was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Several application formulations were prepared and representative analytical samples were collected and dispatched to the analytical laboratories internally. The test item concentrations were determined by HPLC coupled to an ELSD detector and quantified with the area under the peak.

Duration of treatment / exposure:

Males: minimum of 4 weeks (2 weeks prior to mating and 2 weeks of mating)
Females: approximately 7 weeks (2 weeks prior to mating up to day 4 post partum)

Frequency of treatment:

daily, 7 days/week

Remarks:

Doses / Concentrations:
43 (group 2), 160 (group 3), 600 (group 4) mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dosage levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported

BODY WEIGHT: Yes
- Time schedule for examinations: daily from treatment start to day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE:
- was recorded for male animals weekly during pre-pairing, for females weekly in during the pre-pairing period, gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum; food consumption was not recorded during the pairing period

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on day before or day of necropsy for males, for females on day 5 post partum
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 5 males, 5 females
- Parameters examined: Complete Blood Cell Count, Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Leukocyte count (total), Differential leukocyte count, Platelet count, Coagulation, Prothrombin time (= Thromboplastin time), Activated partial Thromoplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on day before or day of necropsy for males, for females on day 5 post partum
- Animals fasted: Yes
- How many animals: 5 males, 5 females
- Parameters examined: Glucose, Urea, Creatinine, Bilirubin (total), Cholesterol (total), Triglycerides, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Gamma-glutamyl-transferase, Bile acids, Creatine kinase, Sodium, Potassium, Chloride, Calcium, Phosphorus, Protein (total), Albumin, Globulin, Albumin/Globulin ratio

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: males shortly before the scheduled sacrifice and females on day 3 or 4 post partum
- Dose groups that were examined: each group
- Battery of functions tested: sensory activity / grip strength / motor activity / other: cage-side and hand-held observations

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

ORGAN WEIGHTS
- testes and epididymides of all parental males were weighed as pairs, from 5 males and females selected randomly from each group, the following organs
were trimmed from any adherent tissue, as appropriate, and their wet weight taken: Adrenal glands (weighed as pairs), Brain, Heart, Kidneys (weighed as pairs), Liver, Thymus, Spleen

HISTOPATHOLOGY: Yes
- the following tissues from all parental males were preserved in neutral phosphate buffered 4% formaldehyde solution:
Prostate, Seminal vesicles with coagulating gland, Testes (in Bouin’s fixative), Epididymides (in Bouin’s fixative)
- the following tissues from all parental females were preserved in neutral phosphate buffered 4% formaldehyde solution:
Ovaries
- In addition, from the five males and females per group selected for organ weights, the following tissues were preserved in neutral phosphate buffered 4% formaldehyde solution:
Gross lesions, Brain (cerebrum, cerebellum, pons), Spinal chord, Small and large intestines (incl. Peyer’s patches), Stomach, Liver, Kidneys, Adrenals, Spleen, Heart, Thymus, Thyroid and Parathyroid, Trachea and lungs (preserved by inflation with fixative and then immersion), Uterus (with vagina), Urinary bladder, Lymph nodes (mandibular and mesenteric), Peripheral nerve (sciatic), Bone marrow

Statistics:

The following statistical methods may be used to food consumption, body weight, macroscopic findings, organ weights and reproduction data:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test was applied to the macroscopic findings.

Details on results:

CLINICAL SIGNS AND MORTALITY
One female in group 2 died accidentally after blood sampling. This death was not considered to be test item-related.
In group 4, all animals were noted to push the head through the bedding material starting on day 9 of the pre-pairing period onwards. One female had salivation after administration of the test item on day 9 of the gestation period. These signs are considered to be signs of discomfort and not to be adverse. In groups 2 and 3, no clinical signs or observations were noted.

BODY WEIGHT AND BODY WEIGHT GAIN
- Males - pre-pairing period: In group 4, mean body weight gain was statistically significantly decreased during the prepairing period (+6.6% versus +13.7% in the control group). Although this decrease had no statistically significant impact on the mean body weight, it was considered to be a test item related effect. During the pairing period, mean body weight and mean body weight gain were not affected by treatment with the test item. In groups 2 and 3, mean body weight and mean body weight gain were not affected by the treatment with the test item for the entire duration of the study. In group 3, mean body weight gain was statistically significant lower on day 6 of the pre-pairing period however, statistical significance occurred on a single day only and therefore it was not considered to be an adverse effect. During the pairing period, mean body weight gain was lower (+8.2% versus +12.0% in the control group) attaining statistically significance on days 9, 13 and 14. However, this was considered to be incidental as there was no-dose dependency.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean body weight gain was statistically significant decreased during the prepairing period (+3.5% versus +8.2% in the control group). This was considered to be a test item related effect, although it did not affect the mean body weight. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 3, mean body weight gain was lower during the pre-pairing period (+5.0% versus +8.2% in the control group) attaining statistical significance between day 6 and 8 and on days 11
and 14. This was considered an effect of the test item although mean body weight was not affected. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 2, mean body weight and mean body weight gain were not affected for the entire duration of the treatment with the test item.

FOOD CONSUMPTION
- Males - pre-pairing period: In group 4, mean food consumption was statistically significant reduced between days 1-8 of the pre-pairing period (-19.2% compared to the control group) and it remained lower between days 8-14 (-10.6% compared to the control group) without attaining statistical significance. In group 3, mean food consumption was not considered to be affected by the treatment with the test item. As mean food consumption was slightly lower between days 1 and 8 of the pre-pairing period (-4.9% compared to the control group) and slightly higher between days 8 and 14 of the pre-pairing period (+2.1% compared to the control group), these variations were considered to be incidental. In group 2, no test item-related effects were noted.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean food consumption was statistically significant decreased between days 1-8 of the pre-pairing period (-15.6% compared to the control group). This was considered to be a test item-related effect. During the gestation and lactation periods mean food consumption was not considered to be affected by treatment with the test item. The statistically significant higher food consumption during the lactation period (+23.0% compared to the control group) was considered to be incidental. In group 3, mean food consumption was decreased between days 1-8 of the pre-pairing period (-10.8% compared to the control group). This was considered to be a transient test item-related effect even though it was not statistically significant. During the gestation and lactation periods, no test item-related effects were noted. In group 2, mean food consumption was not affected by treatment with the test item. During the lactation period, the statistically significant higher mean food consumption was considered to be incidental.

HAEMATOLOGY
- Males: In group 4, the absolute count of neutrophils was statistically significant higher (+40.6% compared to the control group). Since the relative count was not statistically significant increased, this was not considered to be adverse. Mean corpuscular hemoglobin concentration was slightly but statistically significant decreased (-2.8% compared to the control group). Since the mean corpuscular hemoglobin was not affected, it was not considered to be an adverse
effect. In group 3, the statistically significant higher relative red cell volume distribution width (+19.0% compared to the control group) was within the range of historical reference value. In group 2 no changes were noted.
- Females: The assessment of the hematology data did not reveal any test item-related effects in females.

CLINICAL CHEMISTRY
- Males: In group 4, urea and potassium concentrations were statistically significant increased (+25.0% and +15.2%, respectively, compared to the control group). These alterations correlate with the histopathological findings noted in the kidney, thus they were considered to be test item-related. In group 3, the statistically significant lower phosphorus concentration (-13.6% compared to the control group) was within the range of historical reference values. The statistically significant lower concentration of total bilirubin in groups 2 and 3 (-27.6% and -26.8% compared to the control group, respectively) was not considered to be a test item-related effect because there was no dose-dependency.
- Females: The assessment of the clinical biochemistry parameters did not reveal any test item-related effect.

NEUROBEHAVIOUR
- Functional Observational Battery: There was no indication of any test item-related effect during the open field phase. Mean values of grip strength (fore- and hind paws) and landing foot splay gave no indication of any test item-related effects. In groups 3 and 4, mean body temperature was statistically significant reduced in males compared to the control group (37.5°C and 37.4°C, respectively, compared to 38.6 °C in the control group).
- Locomotor Activity: Locomotor activity was assessed quantitatively in terms of low beam counts in an activity monitor. In all groups there was no indication of a test item-related effect. The statistically significant lower locomotor activity noted at 30 min in group 4 males, and the statistically significant lower total locomotor activity noted in group 4 females were within the range of the historical control data.

ORGAN WEIGHTS
- Males: In group 4, mean relative liver and kidney weights were statistically significant increased.
- Females: In group 4, mean absolute and relative liver weights were statistically significant increased.
These findings correlated with the histopathological findings and were therefore considered to be test item-related.

GROSS PATHOLOGY
The findings noted during the macroscopic examinations did not give any indication of test item related effects.
- Males: In group 4, one male was noted to have reddish foci on the thymus, a second male was noted to have a dark red discoloration of the mandibular lymph node and a third male to have enlarged liver. Dark red or reddish foci were noted on the thymus of one male in group 3, in three males in group 2 and in one male in group 1. One male in group 1 was noted to have a subcutaneous yellowish-soft nodule on the right side of thoracic dorsal region.
- Females: In group 4, an isolated reddish focus in the lung of one female and dark red foci in the stomach of a second female were noted. In group 3, one female was noted to have a dark brown content in the stomach, jejunum and ileum. Dark red foci were also noted on the mucosa of fundus. The other findings noted were dark red discoloration of both ovaries in a second female and enlargement of the renal lymph nodes in a third female. In group 2, dark red foci were noted on the thymus in two females (one of these females died after the blood sampling).

HISTOPATHOLOGY:
- Liver: Minimal centrilobular hepatocellular hypertrophy was observed in four males in group 3 and slight centrilobular hepatocellular hypertrophy in one male in group 4; minimal to slight diffuse hepatocellular hypertrophy was noted in the remaining four males and all five females in group 4. Both types of liver cell hypertrophy were considered to represent an adaptive response to the increased need for metabolizing the test item.
- Thyroid gland: In group 4, minimally increased incidence and severity of diffuse follicular cell hypertrophy was noted in males and females. This change was considered to be secondary to the enhanced liver cell metabolism due to hepatocellular hypertrophy.
- Stomach: Minimal acanthosis of nonglandular stomach was present in all males and in three females in group 4 and was associated with minimal to slight focal/multifocal chronic inflammation in two of these males. Isolated, slight and multifocal chronic inflammation occurred in two females in group 3 and one male in group 4. Minimal multifocal chronic inflammation in the glandular stomach was noted in one male in group 3, and slight focal chronic inflammation in one female in group 3. The findings in the female were associated with minimal and multifocal congestion. Slight multifocal congestion occurred in two females in group 4, in one female associated with an acute thrombus and in the other with minimal and multifocal chronic inflammation. All of the stomach findings were considered to represent a local irritating effect of the test item.
- Kidneys: Minimally increased incidence of hyaline droplets/granules was observed in males in groups 3 and 4 (five males in each group, compared to three in the control group). These eosinophilic hyaline droplets/granules, which reflected an increased content of α-2μ-globulin, occur within the cytoplasm of proximal convoluted tubules of sexually mature male rats. Because a slightly increased severity was also noted (mean severity of 2.2 compared to 1.0 in the control males), this was considered to be an adverse effect of the test item in group 4 (600 mg/kg) males.
- Spleen: Extramedullary hematopoiesis was minimally increased in severity in females in group 4, possibly secondary to or an adaptation to stress and/or impaired health conditions. In group 4, the minimally increased incidence of thymus atrophy in females was also considered to be secondary to stress rather than to represent a direct effect of the test item.
All other microscopic findings noted in various organs and in all groups examined were considered to be incidental in nature because their morphology, severity, and incidence did not distinguish treated rats from controls.

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: statistically significant reductions in food consumption and body weight gain, histopathological findings and clinical biochemistry changes, reduced body temperature (in males)

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: effects on kidneys (histopathology + clinical biochemistry), food consumption and body weight gain

Critical effects observed:

not specified

Table 1: Differences in mean Food consumption (Males in g/animal/day), pre-pairing period

Group	Days
	1 – 8		8 - 14		1 - 14
mg/kg	g	(%)*	g	(%)*	g	(%)*
1 (0)	24.5		23.5		24
2 (43)	24.3	-0.8	25.6	+8.9	25	+4.2
3 (160)	23.3	-4.9	24	+2.1	23.7	-1.3
4 (600)	19.8	-19.2	21	-10.6	20.4	-15.0

*percentages refer to the values of group1

 

Table 2: Differences in mean Body weight gain males, pre-pairing period

Group	Days
	1 – 8		8 - 14		1 - 14
mg/kg	g	(%)*	g	(%)*	g	(%)*
1 (0)	22	+7.0	21	+6.3	43	+13.7
2 (43)	20	+6.3	23	+6.8	43	+13.5
3 (160)	16	+5.0	17	+5.1	33	+10.4
4 (600)	10	+3.1	11	+3.3	21	+6.6

*increase within the respective time interval

 

Table 3: Differences in mean Food consumption (Females in g/animal/day), pre-pairing period

Group	Days
	1 – 8		8 - 14		1 - 14
mg/kg	g	(%)*	g	(%)*	g	(%)*
1 (0)	16.7		15.3		16
2 (43)	16.6	-0.6	16.7	+9.2	16.6	+3.8
3 (160)	14.9	-10.8	16.3	+6.5	15.6	-2.5
4 (600)	14.1	-15.6	15.7	+2.6	14.9	-6.9

*percentages refer to the values of group1

 

Table 4: Differences in mean Body weight gain females, pre-pairing period

Group	Days
	1 – 8		8 - 14		1 - 14
mg/kg	g	(%)*	g	(%)*	g	(%)*
1 (0)	5	+2.6	11	+5.5	16	+8.2
2 (43)	6	+3.0	7	+3.4	13	+6.4
3 (160)	0	+0.0	10	+5.0	10	+5.0
4 (600)	1	+0.5	6	+3.0	7	+3.5

*increase within the respective time interval

 

Table 5: Clinical Biochemistry

		Group 1	Group 2	Group 3	Group 4
Days (pre-pairing period)		0	43 mg/kg bw	160 mg/kg bw	600 mg/kg bw
Male (before necropsy)
Urea	MEAN	6.16	5.93	6.79	7.70*
mmol/L	% of control	100	96	110	125
	SD	0.63	0.93	0.72	1.16
	N	5	5	5	5
Potassium	MEAN	4.09	4.27	4.34	4.71*
mmol/L	% of control	100	104	106	115
	SD	0.39	0.24	0.18	0.49
	N	5	5	5	5
Female (day 5 post partum)
Urea	MEAN	9.34	8.16	8.03	8.79
mmol/L	% of control	100	87	86	94
	SD	1.82	1.28	0.63	2.12
	N	5	5	5	5
Potassium	MEAN	4.10	4.22	3.92	3.77
mmol/L	% of control	100	103	96	92
	SD	0.59	0.80	0.53	0.50
	N	5	5	5	5

Significant different compared to control value, *p<0.05

 

Table 6: Organ/body weight ratios

		Group 1	Group 2	Group 3	Group 4
Days (pre-pairing period)		0	43 mg/kg bw	160 mg/kg bw	600 mg/kg bw
Male
Liver (%)	MEAN	2.57	2.53	2.63	3.02**
	SD	0.1	0.07	0.16	0.17
	N	5	5	5	5
Kidneys (%)	MEAN	0.61	0.61	0.61	0.68**
	SD	0.02	0.02	0.03	0.02
	N	5	5	5	5
Female
Liver (%)	MEAN	3.18	3.35	3.19	3.93**
	SD	0.21	0.07	0.25	0.23
	N	5	5	5	5
Kidneys (%)	MEAN	0.63	0.65	0.61	0.67
	SD	0.06	0.02	0.03	0.05
	N	5	5	5	5

Significant different compared to control value, **p<0.01

Conclusions:

Based on the statistically significant reductions in food consumption and body weight gain observed in males and females at 600 mg/kg/day and the transient effects in females at 160 mg/kg/day, on histopathological findings and clinical biochemistry changes observed in males at 600 mg/kg, and on reduced body temperature recorded in males at 160 and 600 mg/kg/day, the systemic NOEL for Sodium coco β-iminodipropionate (CAS 3655-00-3) was established at 43 mg/kg bw/day. The NOAEL can be set at 160 mg/kg bw because at this dose level on transient changes in food consumption and bodyweight were observed in females and slightly reduced body temperature was noted in males without clear toxicological relevance.

Executive summary:

The purpose of this study was to generate information concerning the effects of Sodium cocoβ- iminodipropionate (CAS # 3655-00-3) on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time. In addition, it provides information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

Sodium cocoβ-iminodipropionate (CAS 3655-00-3) was administered to male rats for at least 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. The following dosage levels were administered:

Group 1: 0 mg/kg body weight/day (control group)

Group 2: 43 mg/kg body weight/day

Group 3: 160 mg/kg body weight/day

Group 4: 600 mg/kg body weight/day

Dose levels were adjusted for content of the active ingredient (30.89%) and its purity (87%), applying a correction factor of 3.72. A standard dosage volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (highly purified water).

No test item-related mortality occurred during the study. The death of one female at 43 mg/kg/day after blood sampling was considered to be incidental.

Only signs of discomfort (i.e. pushing the head through the bedding material after administration, a single occasion of salivation) in animals treated at 600 mg/kg/day, but no test item-related clinical signs were observed.

At 160 and 600 mg/kg/day, mean body temperature was statistically significantly reduced in males. Locomotor activity was not affected by treatment with the test item.

 

For males at 600 mg/kg/day, mean food consumption was statistically significantly reduced on days 1-8 of the pre-pairing period.

For females at 160 and 600 mg/kg/day mean food consumption was dose-dependently reduced during the first week of the pre-pairing period.

 

At 160 and 600 mg/kg/day body weight gain was dose dependently decreased during the pre-pairing period in males and female. The decrease was statistically significant at 600 mg/kg/day for males and 160 and 600 mg/kg/day for females. Because body weight was only marginally affected for males at 160 mg/kg/day, the effect at this dosage level was not considered to be an adverse effect.

 

At 600 mg/kg/day, urea and potassium concentrations were statistically significantly increased in males. These alterations were considered to be possibly test item-related as they correlated with a slightly increased incidence and severity of hyaline/droplets in the kidneys.

 

At 600 mg/kg/day, relative kidney weight was increased in males. This finding was considered to be test item-related as it correlated with the histopathological findings in the kidney and the clinical biochemistry changes. Absolute and relative liver weights were statistically significantly increased in males and females. According to the histopathological evaluation that detected hepatocellular hypertrophy, the effect on liver weights was considered to be an adaptive response.

 

The findings noted during the macroscopic examinations did not provide any indication of any test item-related effects.

At 160 and 600 mg/kg/day, the liver cell hypertrophy was considered to represent an adaptive response to an increased need for metabolizing the test item.

The minimally increased incidence and severity of diffuse follicular cell hypertrophy observed in the thyroid gland of males and females treated at 600 mg/kg/day was considered to be secondary to the enhanced liver cell metabolism.

The stomach lesions, observed in both the forestomach (acanthosis associated with chronic inflammation) and the glandular stomach (focal/multifocal chronic inflammation) of several animals treated at 160 and 600 mg/kg/day were considered to be related to the local irritating effect of the test item.

The minimal increase in incidence of hyaline droplets/granules at 160 mg/kg/day was considered to be within the biological range. Because this finding, reflecting an increased content of α-2μ-globulin within the cytoplasm (phagolysosomes) of proximal convoluted tubules of sexually mature male rats, was accompanied by a slight increase in severity in males at 600 mg/kg/day, it was considered to be an adverse effect at this dosage level.

 

Based on the statistically significant reductions in food consumption and body weight gain observed in males and females at 600 mg/kg/day and the transient effects in females at 160 mg/kg/day, on the histopathological findings and clinical biochemistry changes in males at 600 mg/kg/day, and on reduced body temperature recorded in males at 160 and 600 mg/kg/day, the systemic NOEL (No Observed Effect Level) was established at 43 mg/kg body weight/day. The NOAEL can be set at 160 mg/kg bw because at this dose level only transient changes in food consumption and bodyweight were observed in females and slightly reduced body temperature was noted in males without clear toxicological relevance.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

160 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD test guideline and GLP-compliant study, of high reliability (Klimisch 1)

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

A study was conducted to generate information concerning the effects of sodium coco β- iminodipropionate (CAS # 3655-00-3) on the possible health hazards likely to arise from repeated oral exposure and provide information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition, according to OECD 422 test guideline and in compliance with GLP, resulting in high reliability. Administered dose levels were 0 (control group), 43, 160 and 600 mg/kg bw/day.

Based on statistically significant reductions in food consumption and body weight gain in males and females at 600 mg/kg/day and transient effects in females at 160 mg/kg/day, on histopathological findings and clinical biochemistry changes in males at 600 mg/kg/day, and on reduced body temperature recorded in males at 160 and 600 mg/kg/day, the systemic NOEL was established at 43 mg/kg body weight/day. The dose of 160 mg/kg bw can be considered a NOAEL because at this dose level only transient changes in food consumption and bodyweight were observed in females and slightly reduced body temperature was noted in males without clear toxicological relevance.

This highly reliable subacute study conducted on sodium coco β- iminodipropionate was effectively used for risk characterisation using appropriate and conservative assessment factors (for details see 7. Toxicological Information Endpoint Study Summary), because a longer term study appears to be unjustified according to Annex XI Regulation (EC) No 1907/2006 chapter 1.2 "weight of evidence .... Where sufficient weight of evidence for the presence or absence of a particular dangerous property is available:.....—further testing on vertebrate animals for that property shall be omitted, ... ", and also in line with the Title III 'Data sharing and avoidance of unnecessary testing' of Regulation (EC) No. 1907/2006.

There is a 91-day percutaneous toxicity study in rabbits performed for cosmetic purpose available and reported in a US Cosmetic Ingredient Report (CIR) on sodium lauriminodipropionate. As this study is considered not to be reliable, it was not integrated into the present dossier.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Only one highly reliable study available using the oral route.

Repeated dose toxicity: via oral route - systemic effects (target organ) urogenital: kidneys

Justification for classification or non-classification

As the lowest NOAEL obtained was 160 mg/kg bw/day (essentially with findings in kidneys) wich is above the classification reference value (100mg/kg bw for Cat 2) a classification according to GHS (Regulation (EU) 1272/2008) as specific target organ toxicity or an additional classification according to DSD (67/548/EEC) as harmful R48 is not indicated or justified.

Labelling for specific target organ toxicity or danger at prolonged exposure:

GHS:no additional labelling

DSD: no additional labelling