Reaction mass of Benzene sulfonic acid, hexadecyl(sulfophenoxy)-,disodium salt and Benzene sulfonic acid, - oxibis[hexadecyl]-, disodium salt

Carcinogenicity

Key value for chemical safety assessment

Carcinogenicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

carcinogenicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This study was conducted prior to GLP and test guidelines, but sufficient data is available for interpretation of results.

Justification for type of information:

Please see category approach document for further details.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Beagle dogs were divided into matched .groups and started on diets containing 0.0 (control), 1.0, 0.5, 0.25, or 0.125
percent Benax 2A1. Four males and four females comprised each group, except for the 1.0 percent level, which consisted of two
males and four females. The percentage levels given above were approximately equivalent to the administration of 0, 319, 128,
65, and 34 mg/kg/day Benax 281, respectively, over the two-year period.

General appearance and behavior, growth, food consumption, hematological values, determinations of serum urea nitrogen content, alkaline
phosphatase and transaminase activity and bromsulfophthalein dye retention, final organ and body weights, and gross and microscopic examination of the tissues were conducted.

GLP compliance:

no

Species:

dog

Strain:

Beagle

Sex:

male/female

Details on test animals or test system and environmental conditions:

Male and female beagle hounds of approximately three months of age were obtained from a commercial kennel and housed at the Biochemical
Research Laboratory. The pups remained in the laboratory three months prior to the beginning of the experiment, during which time
they were vaccinated for distemper, hepatitis, and leptospira. The stock diet for the first two months was Famo Labomtory Chow; it was then changed to Purina Laboratory Chow for the remaining time. Dogs of each sex per level were housed together and had free access to food and water at a l l times.
On October 17, 1961, the dogs were divided into matched .groups

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

other: Not applicable

Vehicle:

unchanged (no vehicle)

Details on exposure:

Dogs were divided into matched .groups and given diets containing 0.0 (control), 1.0, 0.5, 0.25, or 0.125 percent Benax 2A1. Four males and four females comprised each group, except for the 1.0 percent level, which consisted of two males and four females. The percentage levels given above were approximately equivalent to the administration of 0, 319, 128, 65, and 34 mg/kg/day Benax 281, respectively, over the two-year
period.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

2 years

Frequency of treatment:

Daily

Post exposure period:

None

Remarks:

Doses / Concentrations:
0.0 (control), 1.0, 0.5, 0.25, or 0.125 percent Benax 2A1.
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
0, 319, 128, 65, and 34 mg/kg/day Benax 281.
Basis:
nominal in diet

No. of animals per sex per dose:

Four males and four females comprised each group, except for the 1.0 percent level, which consisted of two males and four females. The

Control animals:

yes, plain diet

Details on study design:

Beagle dogs were divided into matched .groups and started on diets containing 0.0 (control), 1.0, 0.5, 0.25, or 0.125
percent Benax 2A1. Four males and four females comprised each group, except for the 1.0 percent level, which consisted of two
males and four females. The percentage levels given above were approximately equivalent to the administration of 0, 319, 128,
65, and 34 mg/kg/day Benax 281, respectively, over the two-year period.

Each dog was weighed weekly for the first three months of the experiment and twice a week thereafter. Food consumption was recorded
3 during the first , 12th, and 16th months, and one week out of each month from 18 months to the end of the experiment. Hematological
studies and determinations of serum urea nitrogen content and alkaline phosphatase activity were made before the beginning of the experiment and at three, six, nine, 12, 18, and 24 months. Transaminase activity (SGPT) was determined at one and two years and bromsulfophthalein dye retention at one year. Urea nitrogen content and transaminase and alkaline phosphatase activity was determined. Pre-exposure liver biopsies were performed on one dog of each sex per level, as well as at six, 12, and 18 months on the experiment.

At the end of the two-year period the animals were fasted overnight and weighed before examination at autopsy. The lungs, heart, liver,
kidney, spleen, testes, and brain were removed and weighed. Portions of each organ,as well as spinal cord, peripheral nerve, pituitary,
thyroid, adrenal, aorta, lymph node, thymus, esophagus, stomach, small and large intestine, pancreas, urinary bladder, ovary, uterus,
and skeletal muscle were preserved. The tissues were then sent to the International Research and Development Corporation in Mattawan,
Michigan,for preparation of hematoxylin-eosin stained sections and microscopic examination.

Positive control:

None

Observations and examinations performed and frequency:

Each dog was weighed weekly for the first three months of the experiment and twice a week thereafter. Food consumption was recorded
3 during the first , 12th, and 16th months, and one week out of each month from 18 months to the end of the experiment.

Sacrifice and pathology:

Hematological studies and determinations of serum urea nitrogen content and alkaline phosphatase activity were made before the beginning of the experiment and at three, six, nine, 12, 18, and 24 months. Transaminase activity (SGPT) was determined at one and two years and bromsulfophthalein dye retention at one year. Urea nitrogen content and transaminase and alkaline phosphatase activity was determined. Pre-exposure liver biopsies were performed on one dog of each sex per level, as well as at six, 12, and 18 months on the experiment.

At the end of the two-year period the animals were fasted overnight and weighed before examination at autopsy. The lungs, heart, liver,
kidney, spleen, testes, and brain were removed and weighed. Portions of each organ,as well as spinal cord, peripheral nerve, pituitary,
thyroid, adrenal, aorta, lymph node, thymus, esophagus, stomach, small and large intestine, pancreas, urinary bladder, ovary, uterus,
and skeletal muscle were preserved. The tissues were then sent to the International Research and Development Corporation in Mattawan,
Michigan,for preparation of hematoxylin-eosin stained sections and microscopic examination.

Other examinations:

None

Statistics:

No data

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

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

Urinalysis findings:

not examined

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:

Benax 2A1 was fed in the diets of male and female beagle hounds for a period of two years in the concentrations of 1.0, 0.5, 0.25, or 0.125 percent. Food congumption data indicated that the dietary concentrations given above administered the test substance in amounts of 319, 128, 65 and 34 mg/kg/day respectively. No evidence of adverse effect whatsoever was observed in the dogs on the 0.5 percent level or below as judged by general
appearance and behavior, growth , food consumption, hematological values, determinations of serum urea nitrogen content, alkaline phosphstase and transaminase activity, and bromsulfophthalein dye retention, final body and organ weights, and gross and microscopic examination of the tissue.

The two males and four females which received 1.0 percent Benax 2A1 in their diets showed growth retardation. Because male dog #330 and female dog #343 lost approximately one-third of their original weights, they were sacrificed after 14 months of the experiment. The final weight of the other male was essentially the same as his original weight, while two of the remaining females lost weight and the third gained. The 1.0% level was not readily acceptable to the dogs. Persistent scratching at the feeders was noted during the early months of the experimental period. This was noted to some extent also in the group of dogs receiving the 0.5% level. Food consumption records for the first month reflect this observation in that the unusually high figures in comparison with the controls was due to spillage and wastage. Therefore, it is possible that the growth retardation in the dogs maintained on the diet containing 1.0 percent Benax 2A1 may be related somewhat to decreased food intake, at least during the early part of the experiment.Intestinal irritation , as evidenced by loose stools and diarrhea which these dogs exhibited for the first 45 days on the experiment, also may have contribtoted to their failure to gain weight.

Alkaline phosphatase determinations at various intervals throughout the two-year period showed a slight increase in activity in both of the male dogs and in two of the four females on the 1.0 percent level. However, the determination of two other liver function tests , that of bromsulfophthalein dye
retention at approximately one year and transaminase activity at one and two years, gave normal results in comparison with the control values.]

An increased liver/body weight ratio was found in the male dog which was sacrificed after 14 months. The organ/body weight ratio of the kidney of the 1.0 percent male which was carried through to the end of the two-year period was also increased. However, these variations are due to decreased body weights, since there was no increase in the organ weights when considered on the absolute basis in grams.

Hematological values, serum urea nitrogen determinations, and gross and microscopic examination of the tissues gave no indication of any adverse effects in the dogs which received 1.0 percent Benax 2A1 in their diets when compared with the controls.

Relevance of carcinogenic effects / potential:

No significant lesions were observed in the tissues of dogs given diets containing 1.0% Benax 2A1.

Dose descriptor:

NOEL

Effect level:

128 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: Based on growth retardation, loose stools and diarrhea, and increased alkaline phosphatase observed in dogs given 319 mg/kg/day or 1.0% in diet. Note: 128 mg/kg/day= 0.5% on diet;

Remarks on result:

other:

Remarks:

Effect type: other: toxicity/carcinogenicity (migrated information)

None

Conclusions:

The NOEL in Beagle dogs was determined to be 0.5% or 128 mg/kg/day.

Executive summary:

Male and female beagle hounds were maintained for two years on diets containing 0.5, 0.25, or 0.125 percent Benax 2A1 without evidence of adverse effect as judged by general appearance and behavior, growth, food consumption, hematological values, determinations of serum urea nitrogen content, alkaline phosphatase and transaminase activity and bromsulfophthalein dye retention, final organ and body weights, and gross and microscopic examination of the tissues. Food consumption data indicated that these dogs indicated that these dogs ingested the test material in amounts of 128, 65, and 34 mg/kg/day, respectively.

Growth was retarded in the male and female dogs which received 1.0 percent (319 mg/kg/day) Benax 2A1 in their diets. These dogs also had loose stools and diarrhea for the first 45 days of the experiment. Alkaline phosphatase activity was increased slightly in both of the male dogs and in two out of four female-dogs on this level. Although there were variations in a few organ/body weight ratios, there was no difference in organ weights when considered on the absolute basis in grams. Hematological determinations, serum urea nitrogen and transaminase values, determination of bromsulfophthalein dye retention, and gross and microscopic examination of the tissues gave no indication of adverse effects when compared with the controls.

Reference 2

Endpoint:

carcinogenicity: oral

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This study was conducted prior to GLP and test guidelines, but sufficient data is available for interpretation of results.

Justification for type of information:

Please see category approach document for further details.

Qualifier:

no guideline followed

Principles of method if other than guideline:

Groups of male and female rats were maintained for a period of two years on diets containing 1.0, 0.3, 0.1, or 0.03 percent Benax 2A1. On a body weight basis, the test substance was consumed in the amounts of 500, 150, 50, or 15 mg/kg/day by the rats on these levels. General appearance and behavior, mortality, incidence of tumorous growths, food consumption, hematological studfes, serum urea nitrogen and alkaline phosphatase determinations, bone marrow examination, final average body and organ weights, and gross and microscopic examination of the tissues were conducted.

GLP compliance:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

Weanling rats from the stock colony of the Biochemical Research Laboratory were placed on a diet of ground Famo Chow and observed
for a period of about four weeks before being divided into well matched groups according to body weight of 30 of each sex per
group. As many as ten males and ten females in each group were designated to be sacrificed for examination after 12 and 18 months
on the experiment. When approximately 50 days of age, the groups of rats were started on diets containing 0.0 (control, 500, 150,
50, or 15 mg/kg/day)of Benax 2A1. These levels are equivalent to the administration of 1.0, 0.3, 0.1 and 0.03 percent Benax 2A1,
respectively, in the diet of adult rats. For the first five months of the experiment, the percent of chemical in feed was adjusted according to body weight and food intake in order to administer the specified levels on a mg/kg/day basis. Experimental diets were prepared by thoroughly mixing the test material with the basic diet of ground Famo Chow. After five months of the experiment, the stock diet was changed to ground Purina Laboratory Chow. The rats were caged individually and allowed food and water ad libitum.

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

other: Not applicable

Vehicle:

unchanged (no vehicle)

Details on exposure:

Rats were given diets containing 0.0 (control, 500, 150, 50, or 15 mg/kg/day)of Benax 2A1. These levels are equivalent to the administration of 1.0, 0.3, 0.1 and 0.03 percent Benax 2A1, respectively, in the diet of adult rats. For the first five months, the percent of chemical in feed was adjusted
according to body weight and food intake in order to administer the specified levels on a mg/kg/day basis.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

2 years

Frequency of treatment:

Daily

Post exposure period:

None

Remarks:

Doses / Concentrations:
1.0, 0.3, 0.1 and 0.03 percent Benax 2A1 and controls
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
0.0 (control, 500, 150, 50, or 15 mg/kg/day)of Benax 2A1
Basis:
nominal in diet

No. of animals per sex per dose:

30/sex/dose

Control animals:

yes, plain diet

Details on study design:

Weanling rats from the stock colony of the Biochemical Research Laboratory were placed on a diet of ground Famo Chow and observed for a period of about four weeks before being divided into well matched groups according to body weight of 30 of each sex per group. As many as ten males and ten females in each group were designated to be sacrificed for examination after 12 and 18 months on the experiment. When approximately 50 days of age, the groups of rats were started on diets containing 0.0 (control, 500, 150, 50, or 15 mg/kg/day)of Benax 2A1. These levels are equivalent to .
the administration of 1.0, 0.3, 0.1 and 0.03 percent Benax 2A1, respectively, in the diet of adult rats. For the first five months of the experiment, the percent of chemical in feed was adjusted according to body weight and food intake in order to administer the specified levels on a mg/kg/day basis. Experimental dietrs were prepared by thoroughly mixing the test material with the basic diet of ground Famo Chow. After five months of the experiment, the stock diet was changed to ground Purina Laboratory Chow, The rats were caged individually and allowed food and water ad libitum.

During the course of the experiment, the rats were observed frequently for any changes in appearance or behavior. Each rat was weighed twice a week for the first month, weekly for the next five months, and every two weeks to the end of the experiment. Whenever possible, failing animals were autopsied when moribund in an effort to ascertain the cause of impending death. In addition, records were kept of mortality, and food consumption was recorded , during the second month of the experiment.

Hematological studies, including hematocrit, hemoglobin content, white blood cell count, and differential white cell count were made on five male and five female rats from the control, 1.0, and 0.3 percent levels after four months on the experiment. At the end of 6, 9, 12, 18, and 24 months, hematological values were obtained from five rats of each sex from each of the groups.

At the end of the two-year period, all surviving rats were fasted overnight, sacrificed by decapitation and examined grossly at autopsy. The lungs, heart, liver, kidneys, spleen, testes, and brain were removed and weighed. Portions of each organ, as well as adrenal, pancreas, urinary bladder, prostate gland, spinal cord, aorta, thymus, peripheral nerve, large intestine, small intestine, stomach, esophagus, pharynx, and skeletal muscle were
preserved in formalin, and hematoxylin-eosin stained sections were prepared for microscopic examination. Bone marrow smears were prepared from the femurs of male and female rats on each level and stained with Wrights' s stain. Samples of blood serum were obtained for the determination of urea nitrogen content and alkaline phosphatase activity using the Technicon Auto-Analyzer.

This same procedure was followed for the interim sacrifices after 12 and 18 months.

When appropriate, the Fisher " t " test was used in comparing the mean values obtained on the experimental groups with those of the control; in general, probability values (P) of less than 0.05 were interpreted as indicating a significant difference.

Positive control:

None

Observations and examinations performed and frequency:

During the course of the experiment, the rats were observed frequently for any changes in appearance or behavior. Each rat was weighed twice a week for the first month, weekly for the next five months, and every two weeks to the end of the experiment. Whenever possible, failing animals were autopsied when moribund in an effort to ascertain the cause of impending death. In addition, records were kept of mortality, and food consumption was recorded , during the second month of the experiment.

Sacrifice and pathology:

Hematological studies, including hematocrit, hemoglobin content, white blood cell count, and differential white cell count were made on five male and five female rats from the control, 1.0, and 0.3 percent levels after four months on the experiment. At the end of 6, 9, 12, 18, and 24 months, hematological values were obtained from five rats of each sex from each of the groups.

At the end of the two-year period, all surviving rats were fasted overnight, sacrificed by decapitation and examined grossly at autopsy. The lungs, heart, liver, kidneys, spleen, testes, and brain were removed and weighed. Portions of each organ, as well as adrenal, pancreas, urinary bladder, prostate gland, spinal cord, aorta, thymus, peripheral nerve, large intestine, small intestine, stomach, esophagus, pharynx, and skeletal muscle were
preserved in formalin, and hematoxylin-eosin stained sections were prepared for microscopic examination. Bone marrow smears were prepared from the femurs of male and female rats on each level and stained with Wrights' s stain. Samples of blood serum were obtained for the determination of urea nitrogen content and alkaline phosphatase activity using the Technicon Auto-Analyzer.

This same procedure was followed for the interim sacrifices after 12 and 18 months.

Other examinations:

None

Statistics:

When appropriate, the Fisher " t " test was used in comparing the mean values obtained on the experimental groups with those of the control; in general, probability values (P) of less than 0.05 were interpreted as indicating a significant difference.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

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:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

Groups of male and female rats were maintained for a period of two years on diets containing 1.0, 0,3, 0.1, or 0.03 percent Benax 2A1. On a body weight basis for adult rats, the test substance was consumed in amounts of 500, 150, 50, or 15 mg/kg/day, respectively, by these groups, For the first five months of the experiment, the percentage levels were adgusted to maintain a uniform administration of Benax 2A1 on a mg/kg/day basis.

In general, all the groups of rats appeared normal in appearance and behavior throughout the experimental period. There was no significant difference in food consumption between the control rats and those receiving the test material in their diets. Records of mortality and incidence of tumorous
growths show no relationship to the inclusion of Benax 2A1 in the diet of rats.

Growth was normal for the groups of rats which received 0.3 percent (150 mg/kg/day or below) of the test material in feed. The females on the 1.0 percent (500 mg/kg/day) level began to show growth retardation after six months of the experiment. Slighter growth depression was observed in the male rats on this level.

There was no evidence of adverse effect observed in any of the groups of rats maintained on diets containing Benax 2A1 when compared with the controls as Judged by periodic hematological examinations or determination of serum urea nitrogen content and alkaline phosphatase activity.

Twelve Months - Final average body and organ weight ratios showed no significant differences between groups of male control rats and those receiving the diets containing 1.0, 0.3, 0.1 or 0.03 percent Benax 2A1. The organ/body weight ratios of the kidney and spleen of the females on the 0.1 percent level and of the liver of the 0.1 and 0.03 percent females were decreased when compared with the controls. However, these variations were not seen at the higher dose levels and are not considered to be due to the inclusion of Benax 2A1 in the diet of rats. Gross and microscopic examination of the tissues revealed no significant pathological findings in the animals that received the test substance in feed for one year when compared with the
controls.

Eighteen Months - A decrease in the final average body weight of the group of female rats on the 1.0 percent level which were autopsied after 18 months was the only evidence of any adverse effect noted at this time. Gross and microscopic examination of the tissues again showed no significant changes attributable to the test substance.

Twenty-four Months - Upon gross and microscopic examination of the tissues, no significant changes were seen in the animals receiving 1.0, 0.3, 0.1 or 0.03 percent Benax 2A1 in their diets for two years in comparison with the controls. The final average body weight of the females on the 1.0 percent level was significantly decreased resulting in an increase in the brain/body weight ratio. The increased testes weight of the male rats which received 1.0 or 0.3 percent Benax 2A1 in their diets is not considered to be of practical significance.

Relevance of carcinogenic effects / potential:

Upon gross and microscopic examination of the tissues, no significant changes were seen in the animals receiving 1.0, 0.3, 0.1 or 0.03 percent Benax 2A1 in their diets for two years in comparison with the controls.

Dose descriptor:

NOEL

Effect level:

150 mg/kg bw/day (nominal)

Sex:

male/female

Basis for effect level:

other: NOEL was based on growth depression observed in rats given 500 mg/kg/day or 1.0% in diet. Note: 150 mg/kg bw/day= 0.3% in diet

Remarks on result:

other:

Remarks:

Effect type: other: toxicity and carcinogenicity (migrated information)

None

Conclusions:

The NOEL was determined to be 150 mg/kg bw/day or 0.3% Benax 2A1 in the diet.

Executive summary:

Groups of male and female rats were maintained for a period of two years on diets containing 1.0, 0.3, 0.1, or 0.03 percent Benax 2A1 without evidence of adverse effect as judged by general appearance and behavior, mortality, incidence of tumorous growths, food consumption, hematological studies, serum urea nitrogen and

alkaline phosphatase determinations, bone marrow examination, final average body and organ weights, and gross and microscopic examination of the tissues. On a body weight basis, the test substance was consumed in the amounts of 500, 150, 50, or 15 mg./kg/day by the rats on these levels.

The only evidence of any adverse effect whatsoever was seen in the rats which received 1.0 percent (500 mg./kg./day) Benax 2A1 in their diets. Growth was depressed in the group of female rats on this level, with a statistically significant decrease in the final average body weight at the end of two years. The group of male rats showed a slighter, statistically insignificant growth retardation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Acceptable quality for evaluation and assessment with highest NOAEL for carcinogenicity. Two different species were tested in 2-year oral chronic studies.

Carcinogenicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Carcinogenicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available studies with DOWFAX 2A1 category member, with no evidence of tumor formation up to the highest (1% in the diet) doses in rats and dogs, DOWFAX category members are not expected to be carcinogenic. Therefore, no CLP classification for carcinogenicity is proposed for this substance.

Additional information

Category member DOWFAX 2A1 was administered in diet up to 1% dose level to rats and dogs over 2 year period. No significant differences in lesion incidence were observed in the tissues of dogs and rats given diets containing up to the highest DOWFAX 2A1 dose compared with the respective controls. NOAEL for carcinogenicity is > 1% in the diet (>319 mg/kg bw/day for dogs and >500 mg/kg bw/day for rats).