Benzene, 1,1'-oxybis-, tetrapropylene derivs., sulfonated, sodium salts

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1961-1963

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1963

Report date:

1963

Materials and methods

Test guideline

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% Benax 2A1. Four males and four females comprised each group, except for the 1.0% 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 2A1, 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

Remarks:

pre-GLP

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Benzene, 1,1'-oxybis-, tetrapropylene derivs., sulfonated, sodium salts

EC Number:

601-601-6

Cas Number:

119345-04-9

Molecular formula:

C24H32O7S2Na2

IUPAC Name:

Benzene, 1,1'-oxybis-, tetrapropylene derivs., sulfonated, sodium salts

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

CRI Name: Benzenesulfonic acid: X-(X-Dodecyl) -X-Phenoxy-; sodium salt
Trade name: Benax 2A1
Appearance: Light-colored powder
Purity: 90.03%
Composition: 90.03% active, 3.16% water, 5.19% NaCl, 1.62% Na2SO4, and 1.20 ppm Iron
pH- 10.10 (10% in water)
Source: F. C. Tuttle, Benzene Products Laboratory
Reference: Lot 180, Drum #2
M.W.: 440.6

Test animals

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 all times.

On October 17, 1961, the dogs were divided into matched groups.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Dogs were divided into matched groups and given diets containing 0.0 (control), 1.0, 0.5, 0.25, or 0.125% Benax 2A1. Four males and four females comprised each group, except for the 1.0% 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

Doses / concentrations

Doses / concentrations 1

Remarks:

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

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0, 319, 128, 65, and 34 mg/kg/day Benax 2A1
Basis:
nominal in diet

No. of animals per sex per dose:

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

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% Benax 2A1. Four males and four females comprised each group, except for the 1.0% 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:

Non

Examinations

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

Results and discussion

Results of examinations

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:

not examined

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 at the concentrations of 1.0, 0.5, 0.25, or 0.125%. Food consumption 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 given the 0.5% dietary 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% 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% 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 contributed 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% 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% 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% Benax 2A1 in their diets when compared with the controls.

Effect levels

Dose descriptor:

NOAEL

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.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

None

Applicant's summary and conclusion

Conclusions:

The systemic NOEL in Beagle dogs was determined to be 0.5% or 128 mg DOWFAX 2A1/kg bw/day.

Executive summary:

Male and female beagle hounds were maintained for two years on diets containing 0.5, 0.25, or 0.125% Benax/DOWFAX 2A1 (equivalent to 128, 65, and 34 mg/kg/day) 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 compared to control animals. At the highest dose level (1% or 319 mg/kg/day, study LOAEL), growth was retarded in the male and female dogs fed Benax/DOWFAX 2A1. These dogs also had loose stools and diarrhea for the first 45 days of the experiment; slightly increased alkaline phosphatase activity in both male dogs and in two out of four female dogs; variations in a few organ/body weight ratios with 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. Based on these effects, the systemic NOEL in Beagle dogs was determined to be 0.5% or 128 mg DOWFAX 2A1/kg bw/day.