Dodecylbenzenesulphonic acid, compound with 2-amino-2-methylpropan-1-ol (1:1)

Endpoint summary

Description of key information

In a study according to OECD 422, oral administration of dodecylbenzenesulfonic acid to rats resulted in soft faeces, and squamous cell hyperplasia of stomach in both sexes at 400 mg/kg bw/day, and liquid faeces and soled perineal region, a decrease in body weight and food consumption in males at 400 mg/kg bw/day. In histopathology examination, squamous cell hyperplasia of stomach was observed in both sexes at 200 mg/kg bw/day and forestomach erosion/ulcer was observed in males at 400mg/kg bw/day. Based on these effects the NOAEL value was 100 mg/kg bw/day for male and female rats and the LOAEL value was 200 mg/kg bw/day for male and female rats. From these results, the target organ for oral dosing of dodecylbenzenesulfonic acid was considered to be the stomach.

On AMP oral repeated dose studies are available in dogs (maximum concentrations in diet tested 2.8 and 62.5 mg/kg bw) and rats (90-day diet NOAEL 25 mg/kg bw). The effects seen in both species are in the liver which can be related to an effect on choline synthesis, but a clear species difference becomes apparent with the dog being more sensitive (chronic NOAEL2.8 mg/kg bw).

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Remarks:

the tested substance is DDBSA one of the components of the compound

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well-documented study.

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

1) Test animals- Supplier: Orient Bio Co. Ltd. 143-1, Sangdaewon-dong, Jungwon-ku, Sungnam, Gyunggi-do, 462-120 Korea- Age at study initiation: 7-week-old animals for male and female- No. of animals at receipt: 57 for male and female- Body weights at study initiation: 212.5 – 243.8 g for males and 147.6 -168.6 g for females- Age at the first day of treatment: 8 weeks for male and female- Body weight range at the first day of treatment: 274.2∼311.1 g for males and 175.7∼213.4 g for females- All animals were visually examined on acquisition. Only the animals remained in good physical condition during the 6-day acclimatization in the animal room were selected for the test.
2) Environmental condition- Temperature 23 +/- 3 deg C, relative humidity of 50 +/- 10%; ventilation of 10 to 20 times/hours; light/dark cycle 12 h/12 h- All animals used in this study were cared for in accordance with the principles outlined in the "Guide for the Care and Use of Laboratory Animals", a NIH publication.
3) Monitoring- Room temperature was generally in the range 20 ~ 26 deg C, relative humidity was generally in the range 40 ~ 60%. No significant deviations, which can affect the experiment, were observed.
4) Housing and identification of animals- Equal or less than five for the quarantine and acclimatization- Equal or less than two for the pre-mating, treatment and recovery period5) Diet, water and bedding material- Pelleted maintenance diet and tap water ad libitum; no contaminants (analysed)

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The test article of the highest dose group was mixed with water for injection, and the low dose group's test article was prepared by dilution of that of the highest dose group. The test article solutions were prepared once a day before completion of the analytical method validation, and after completion the test article was formulated over once a week.

Duration of treatment / exposure:

From 2 weeks before mating to the end of the mating period for male (at least 28 days)From 2 weeks before mating to day 4 of lactation including the mating and gestation periods for female- Post exposure period: 15 days in both sexes

Frequency of treatment:

Once daily

Remarks:

Doses / Concentrations:
0, 100, 200, and 400 mg/kg bw/day (Dosing volume 10ml/kg/day)
Basis:
nominal in diet

No. of animals per sex per dose:

10 males and females for 100 and 200 mg/kg bw/day and 16 males and females for 400 mg/kg bw/day (10 was for test group and 6 was for recovery group), 16 males and females for vehicle control (10 was for test group and 6 was for recovery group)

Control animals:

yes

Details on study design:

- Dose levels determined in a pilot toxicity study of dodecylbenzenesulfonic acid in rats- Constant dosage volume of 10 mL/kg bw/day: calculated with Path/Tox system according to the basis of recently measured body weight.- Dosing of both sexes was begun at 2 weeks prior to mating. Dosing was continued in both sexes during the mating period. Males were dosed after the mating period at least until the minimum total dosing period of 28 days had been completed. Daily dosing of the parental females was continued throughout pregnancy and at least up to day 4 post-partum

Statistics:

- Body weights, food consumption, organ weights, and clinical pathology : means the standard deviation of each mean. - Bartlett's test : analyzing for homogeneity of variance- Dunnett's t test : analyzing for the significance of inter-group differences- Analysis of Variance : analyzing for homogeneous data- Kruskal-Wallis test : analyzing for Heterogeneous data- Dunn's Rank Sum test : analyzing for the significance of inter-group differences between the control and treated groups- F test : analyzing the data of recovery groups for homogeneity of variance- Dunnett's t test : analyzing for homogeneous data- Dunn's Rank Sum test : analyzing for the significance of inter-group differences- t test : analyzing for Heterogeneous data- Kruskal-Wallis test : analyzing for the significance of inter-group differences between the control and treated group-Statistical analyses were performed by comparing the different dose groups with the vehicle control group using Path/Tox System. - p<0.05 or p<0.01

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Details on results:

-Clinical signs: Treatment-related clinical signs such as salivation and soft feces were observed in all males of the 400 mg/kg bw/day group during the premating and mating periods. Soiled perineal and liquid feces were observed in 2 males of 400 mg/kg bw/day group during the premating. Salivation and soft feces were observed in 9 and 8 females during the premating and in 8 and 5 during the mating period in the 400 mg/kg bw/day group, respectively. Thin appearance, salivation, staining around mouth, soiled perineal region and soft feces were observed in 1, 10, 1, 1 and 4 females during the gestation period in the 400 mg/kg group, respectively. Thin appearance and salivation were observed in 1 and 10 animals in the 400 mg/kg bw/day group during the lactation period, respectively.

- Body weights and food consumption: In males, a statistically significant decrease was observed on days 8 and 14 of premating. In females, there was no statistically significant changes except a decrease on day 20 of gestation at 400 mg/kg bw/day. In males, a statistically significant decrease in food consumption was observed in the 400 mg/kg bw/day group on days 2 and 9 of the premating, and in females it was observed on day 2 of premating and day 8 of gestation.

- Neurobehavioral evaluation: No treatment-related changes were observed in any of the treatment group.- Urinalysis for males: There were no treatment-related changes for males.

- Hematological test : In hematology test, activated partial thromboplastin time (APTT) was statistically significantly decreased in male of the 400 mg/kg bw/day group. No treatment-related changes were observed in females. In recovery group, a statically significant decrease in RBC count (RBC) was observed in males of the 400 mg/kg bw/day group and an increase in mean corpuscular hemoglobin (MCH) was observed in females.

- Biochemical test: In serum biochemistry test, a statistically significant increase in A/G ratio andalanine aminotransferase (ALT) was observed in males of the 400 mg/kg bw/day group. In females, a statistically significant decrease in blood urine nitrogen (BUN) observed in all treatment groups and an decrease in albumin (ALB) was also observed in the 400 mg/kg bw/day group. In recovery group, a statistically significant decrease in ALT and increase in ALP were observed in females of the 400 mg/kg bw/day group.

- Gross findings: There were no treatment-related changes for all animals

- Organ weights: In males, no a statistically significant changes were observed in any of the treatment groups. In females, a statistically significant increase in absolute weight of ovaries was observed in the 200 mg/kg bw/day group, and a decrease in absolute weight of salivary gland and heart was observed in the 400 mg/kg bw/day groups. The mean weight of ovaries for main group was 0.108 g in compared with vehicle control of 0.093 g and the mean weight of salivary glands for main group was 0.462 g in compared with vehicle control of 0.532 g. Also, the mean weight of heart for main group was 0.816 g in compared with vehicle control of 0.955 g. In recovery groups, a significant decrease in liver and kidney was observed in males the 400 mg/kg bw/day group. No significant differences were observed between vehicle control and test group for organ weights in females.

- Histopathological findings: Squamous cell hyperplasia in stomach was observed in males and females at 200 and 400 mg/kg bw/day. Two cases of minimal forestomach erosion/ulcer were also observed in males at 400 mg/kg bw/day.

Key result

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: local effects to the GI tract

Dose descriptor:

LOAEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: local effects to the GI-tract

Critical effects observed:

not specified

Conclusions:

Oral administration of dodecylbenzenesulfonic acid to rats resulted in soft feces, and squamous cell hyperplasia of stomach in both sexes at 400mg/kg bw/day, and liquid feces and soled perineal region, a decrease in body weight and food consumption in males at 400 mg/kg bw/day. In histopathology examination, squamous cell hyperplasia of stomach was observed in both sexes at 200 mg/kg bw/day and forestomach erosion/ulcer was observed in males at 400mg/kg bw/day. Based on these effects the NOAEL value was 100 mg/kg bw/day for male and female rats and the LOAEL value was 200 mg/kg bw/day for male and female rats. From these results, the target organ for oral dosing of dodecylbenzenesulfonic acid was considered to be the stomach.

Executive summary:

Repeated oral dosing of dodecylbenzenesulfonic acid resulted in soft feces, and squamous cell hyperplasia of stomach in both sexes, and liquid feces and soled perineal region, a decrease in body weight and food consumption, forestomach erosion/ulcer in males at 400 mg/kgbw/day, and squamous cell hyperplasia of stomach in both sexes at 200 mg/kgbw/day.Also,activated partial thromboplastin time (APTT) was statistically significantly decreased in male of the 400 mg/kg bw/day group. Inserum biochemistry test, a statistically significant increase in A/G ratio and alanine aminotransferase (ALT) was observed in males of the 400 mg/kg bw/day group.As a result of organ weight test, no a statistically significant changes were observed in any of the treatment groups in males. A statistically significant increase in absolute weight of ovaries was observed in the 200 mg/kg bw/day group, and a decrease in absolute weight of salivary gland and heart was observed in the 400 mg/kg bw/day groups.

Based on results, all findings were completely or partially reversed during the15daysrecovery period. The target organ for oral dosing of dodecylbenzenesulfonic acid was considered as stomach. The NOAEL and the LOAEL for repeated toxicity of dodecylbenzenesulfonic acid was considered to be 100 mg/kg bw/day and 200 mg/kg bw/day in both sexes, respectively.

Reference 2

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Remarks:

the tested substance is AMP one of the components of the compound

Adequacy of study:

key study

Study period:

5/19/1988 to 7/25/1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: meets generally accepted scientific standards, well-documented, and acceptable for assessment

Remarks:

maximum dose level low and without effects

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 452 (Chronic Toxicity Studies)

GLP compliance:

yes (incl. QA statement)

Remarks:

contains an audit certificate

Limit test:

no

Species:

dog

Strain:

Beagle

Sex:

male/female

Details on test animals or test system and environmental conditions:

Twenty nine male and 29 female beagle dogs were received from Laboratory Research Enterprises, Kalamazoo, MI, and 25 of the healthiest of each sex were used for study. The animals were selected based on criteria of the US FDA (1982). The dogs were 4 months of age at arrival to the testing laboratory. Animals were identified by an assigned identification number on a collar, housed singly for an acclimation period, and were examined by a laboratory veterinarian for general health status. Animals were stratified by body weight, and assigned randomly to their treatment groups.

Route of administration:

oral: feed

Vehicle:

other: test material was dissolved in ethanol and then mixed into the feed

Details on oral exposure:

Animals were dosed via the diet, which was tested by the supplier (Purina) for contaminants. The test article was incorporated into the diet on a weight/weight basis using a premix. Fresh food was prepared with the test article weekly. The animals were offered 400g of the diets daily. Any uneaten food was weighed and recorded.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

1 year

Frequency of treatment:

continuous

Remarks:

Doses / Concentrations:0, 1.1, 11, 110 ppmBasis:nominal in diet

Remarks:

Doses / Concentrations:0, 0.031, 0.31, 2.8 mg/kg bw/dayBasis:nominal in diet

No. of animals per sex per dose:

6/sex/dose

Control animals:

yes, concurrent vehicle

Details on study design:

Animals were dosed via the diet, which was tested by the supplier (Purina) for contaminants. The test article was incorporated into the diet on a weight/weight basis using a premix. Fresh food was prepared with the test article weekly. The animals were offered 400g of the diets daily. Any uneaten food was weighed and recorded. Drinking water was provided throughout the study ad libitum. Six males and six females were assigned to each of 4 dose groups (0, 1.1, 11, 110 ppm).Daily observations were made of the general appearance, behavior, the presence of any signs of toxicity or pharmacologic effects and mortality. Body weights were recorded prior to dosing, and weekly thereafter. The fasting terminal body weight was also recorded. While weighing, the animals were examined for lesions or other signs of toxicity. Each animal had an ophthalmoscopic examination once pre-study, weeks 1, month 3,6,9,12 by a veterinarian.Blood samples were obtained pre-study, and at 3, 6, 9, 12 months via venipuncture of fasted animals. The following parameters were recorded: hematocrit, hemoglobin, erythrocyte count, leukocyte count, platelet count, prothrombin time, MCH, MCHC, and MCV, Calcium, Phosphorous, Chloride, Sodium, Potassium, glucose, serum alkaline phosphatase, serum aspartate aminotransferase, serum analine aminotransferase, gamma glutamyl, transpeptidase, blood urea nitrogen, total protein, albumin, globulin, creatinine, bilirubin, and serum protein. Urine was examined for pH, specific gravity, volume and appearance, protein, glucose, acetone bodies, creatinine, and microscopic sediment.Post-mortem examinations included a complete gross examination of the external surfaces, all orifices, the cranial cavity, the external surface of the brain, the thoracic, pelvic, and abdominal cavities and their viscera, cervical tissues and organs, and the carcass. Two animals per sex per dose were sacrificed at 6 months, with the remainder at 12 months. Organ weights were recorded for the liver, kidneys, testes, thyroids, adrenals, and brain. Tissues that were removed and preserved are the same as current OECD guidelines

Positive control:

no

Observations and examinations performed and frequency:

Daily observations were made of the general appearance, behavior, the presence of any signs of toxicity or pharmacologic effects and mortality. Body weights were recorded prior to dosing, and weekly thereafter. The fasting terminal body weight was also recorded. While weighing, the animals were examined for lesions or other signs of toxicity. Each animal had an ophthalmoscopic examination once pre-study, weeks 1, month 3,6,9,12 by a veterinarian.

Sacrifice and pathology:

Blood samples were obtained pre-study, and at 3, 6, 9, 12 months via venipuncture of fasted animals. The following parameters were recorded: hematocrit, hemoglobin, erythrocyte count, leukocyte count, platelet count, prothrombin time, MCH, MCHC, and MCV, Calcium, Phosphorous, Chloride, Sodium, Potassium, glucose, serum alkaline phosphatase, serum aspartate aminotransferase, serum analine aminotransferase, gamma glutamyl, transpeptidase, blood urea nitrogen, total protein, albumin, globulin, creatinine, bilirubin, and serum protein. Urine was examined for pH, specific gravity, volume and appearance, protein, glucose, acetone bodies, creatinine, and microscopic sediment.Post-mortem examinations included a complete gross examination of the external surfaces, all orifices, the cranial cavity, the external surface of the brain, the thoracic, pelvic, and abdominal cavities and their viscera, cervical tissues and organs, and the carcass. Two animals per sex per dose were sacrificed at 6 months, with the remainder at 12 months. Organ weights were recorded for the liver, kidneys, testes, thyroids, adrenals, and brain. Tissues that were removed and preserved are the same as current OECD guidelines

Statistics:

yes

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Test material intake is estimated based on week 36 body weights and food consumption.Males estimated (mg/kg/day)Control 01.1ppm .03111ppm 0.31110ppm 2.98Females estimated (mg/kg/day)Control 01.1ppm .02911ppm 0.31110ppm 2.55There were no general in-life observations made for any treated or control animal. Likewise, there were no ophthalmoscopic observations noted for any animal. Neither an ANOVA or a Kruskal-Wallis non-parametric ANOVA revealed any statistical significance in food consumption patterns for any dose level in either sex. There was no statistically significant differences in body weights for any of the animals throughout the study. Clinical chemistry findings for males revealed a slight decrease in the low and mid-dose groups' albumin-globulin ratio. Since there was no dose-response (the high dose group was not different than the control), it was judged to be a spurious finding and not treatment-related. High-dose males at 9 and 12 months showed differences in serum albumin via serum electrophoresis, but was not corroborated by actual clinical chemistry measurements, and was not considered by the authors to be biologically-significant. Throughout the study, findings for females included differences in serum sodium, serum ALT, and serum AST, but were transient, did not appear dose-related, and were judged by the authors to be not related to the administration of AMP. There was no effect of AMP administration on hematology, and likewise the urinalysis revealed no treatment-related effects. There were no effects on organ weights or organ/body weight ratios that were attributed to test material administration by the authors.At necropsy, evaluation of the tissues revealed several isolated observations, none of which were attributed to the administration of the test material. There were no neoplastic observations in any of the dogs sacrificed at either 6 months or one year of administration in their diets.

Dose descriptor:

NOAEL

Effect level:

> 110 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Dose equivalent to 2.8 mg/kg bw/day based on average male and female test material consumption

Critical effects observed:

not specified

Conclusions:

Based on the findings under these study conditions, there is no effect at any dose level on general appearance, behavior, body weight, food consumption, ophthalmoscopic exams, clinical chemistry, hematology, organ weights, or tissue histopathology. Based on the absence of statistically and biologically significant findings in dose-response patterns, the No-Observed Effect Level for AMP in the diets of Beagle dogs in greater than 110 ppm (>2.8 mg/kg bw).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

13.3 mg/kg bw/day

Study duration:

chronic

Species:

dog

Quality of whole database:

The NOAEL is based on one of the components in a worst case approach (the substance consists of 21%w/w AMP and 79 %w/w DDBSA). Taking into account only the toxicity of AMP the chronic NOAEL is calculated to be 13.3 mg/kg bw (based on a NOAEL of 2.8 mg/kg bw in dogs). Taking into account in addition the subactute NOAEL for DDBSA of 100 mg/kg bw with the standard safety factor for extrapolation to a sub-chronic NOAEL this leads to an overall NOAEL of 55 mg/kg bw (2.8/0.21 + 33.3/0.79). This last approach is based on addition and is considered less suitable for these components, as the target tissue is the liver for AMP and the GI-tract for DDBSA.

System:

hepatobiliary

Organ:

liver

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

Justification for classification or non-classification

Based on the data on the components, the substance does not need to be classified for repeated dose toxicity according to CLP (Regulation EC No 1272/2008).