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Administrative data

Description of key information

Several GLP and non-GLP studies containing sufficient data for the interpretation and assessment are available, including rat and dog studies on the REACH-registered substance and its category ADPODS analogues.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
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.
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
Species:
dog
Strain:
Beagle
Sex:
male/female
Details on test animals 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.
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
Remarks:
Doses / Concentrations:
0.0 (control), 1.0, 0.5, 0.25, or 0.125% Benax 2A1
Basis:
nominal in diet
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
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:
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.
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.
Critical effects observed:
not specified

None

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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
128 mg/kg bw/day
Study duration:
chronic
Species:
dog
Quality of whole database:
acceptable

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

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting 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.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
no guideline followed
Principles of method if other than guideline:
The skin irritation test included topical application of 0.5 ml of neat Dowfax 2A1 to the ear, and to intact and abraded skin on the abdomen of a male New Zealand White (NZW) rabbit. The animal was bandaged for 24 hours after application of the test material to provide occlusion and prevent grooming of the application site. Twenty four hours after application the bandage was removed and the application sites were graded. Five consecutive daily doses of Dowfax 2A1 were applied to the intact abdominal skin, and the apex of the left pinna, and three consecutive daily applications to the abraded abdominal site. Observations were recorded up to 72 hours after the final dose.
GLP compliance:
no
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male
Details on test animals and environmental conditions:
No additional data
Type of coverage:
other: ear- open; intact and abraded abdomen- occluded
Vehicle:
unchanged (no vehicle)
Details on exposure:
The skin irritation test included topical application of 0.5 ml of neat Dowfax 2A1 to the ear, and to intact and abraded skin on the abdomen of a male New Zealand White (NZW) rabbit. The animal was bandaged for 24 hours after application of the test material to provide occlusion and prevent grooming of the application site. Twenty four hours after application the bandage was removed and the application sites were graded
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not applicable
Duration of treatment / exposure:
24 hours
Frequency of treatment:
Five consecutive daily doses of Dowfax 2A1 were applied to the intact abdominal skin, and the apex of the left pinna, and three consecutive daily
applications to the abraded abdominal site.
Remarks:
Doses / Concentrations:
0.5 ml
Basis:
other: amount applied
No. of animals per sex per dose:
1
Control animals:
no
Details on study design:
The skin irritation test included topical application of 0.5 ml of neat Dowfax 2A1 to the ear, and to intact and abraded skin on the abdomen of a male New Zealand White (NZW) rabbit. The abdomen of the rabbit was shaved with a straight razor at least three days prior to test initiation. The animal was
bandaged for 24 hours after application of the test material to provide occlusion and prevent grooming of the application site. Twenty four hours
after application the bandage was removed and the application sites were graded. Five consecutive daily doses of Dowfax 2A1 were applied to the intact abdominal skin, and the apex of the left pinna, and three consecutive daily applications to the abraded abdominal site. The study was terminated 72 hours after the final dose.
Positive control:
None
Observations and examinations performed and frequency:
Twenty four hours after application the bandage was removed and the application sites were graded. Observations were recorded out to 72 hours after the last dose.
Sacrifice and pathology:
No data
Other examinations:
None
Statistics:
None
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not examined
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:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
The animal survived the test period. No clinical signs indicative of systemic toxicity were observed.

No irritation was observed on the ear. Very slight erythema was observed at the intact site after the last application. Very slight erythema was observed at the abraded site on test day two through test day five. The study was terminated 72 hours after the final dose.
Critical effects observed:
not specified

None

Conclusions:
No irritation was observed on the ear. Very slight erythema was observed at the intact site after the last application. Very slight erythema was observed at the abraded site on test day two through test day five. The study was terminated 72 hours after the final dose.
Executive summary:

The skin irritation test included topical application of 0.5 ml of neat Dowfax 2A1 to the ear, and to intact and abraded skin on the abdomen of a male New Zealand White (NZW) rabbit. Five consecutive daily doses of Dowfax 2A1 were applied to the intact abdominal skin, and the apex of the left pinna, and three consecutive daily applications to the abraded abdominal site. No irritation was observed on the ear. Very slight erythema was observed at the intact site after the last application. Very slight erythema was observed at the abraded site on test day two through test day five. The study was terminated 72 hours after the final dose.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
5.75 mg/cm²
Study duration:
subacute
Species:
rabbit
Quality of whole database:
adequate

Additional information

ORAL STUDIES OVERVIEW:

In the 1963 KS (Klimisch 2), groups of male and female beagle hounds were maintained for two years on diets containing 0, 34, 65, 128 mg/kg/day Benax/DOWFAX 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. Growth was retarded in the male and female dogs fed higest dose of 319 mg/kg/day DOWFAX 2A1 (study LOAEL). These dogs also had loose stools and diarrhea for the first 45 days of the experiment; slightly increased alkaline phosphatase activity in males and half of females; and variations in a few organ/body weight ratios without a difference in organ weights when considered on the absolute basis in grams. Based on all these observations, the general systemic toxicity NOAEL (and point of departure for DNEL derivation) for the dog two year study is 128 mg/kg/day.

In the 1963 ss (Klimisch 2), groups of 30 male and female rats were maintained for a period of two years on diets containing 0, 15, 50, 150, 500 mg DOWFAX 2A1/kg bw/day 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. Based on depressed growth and statistically significant decrease in the final average body weight at the end of two years seen in female rats fed 500 mg/kg/day BOWFAX 2A1 (study LOAEL), the general systemic toxicity NOAEL for the rat two year study is 150 mg/kg/day.

In the 1957 ss (Klimisch=2), rats were administered 0, 0.01, 0.03, 0.1, 0.3 and 1.0 % DOWFAX 2A1 in the diet for 90 days. Livers of male rats at the 1% level showed central lobular necrosis of the parenchymal cells, and fatty degeneration and early, but slight fibrosis with some slight bile duct epithelium proliferation in the portal areas when examined microscopically. Female rats at the 1% level showed a retardation of growth together with a statistically significant increase in the average weight of the liver and the kidneys. Grossly, the livers were light in color with a mottled appearance. Upon microscopic examination, effects similar to those noted above in male rats were observed. Based on the above findings the NOEL was 0.3% in the diet.

In the 1963 ss (Klimisch 2), dogs were administered 0, 40, 131 or 350 mg/kg/day (0, 0.1, 0.3 and 1.0%) DOWFAX 2A1 in the diet for 95 days. The dogs in the 1% or 350 mg/kg/day dose group did not accept their diet to the same degree as the animals on the control or other dietary levels. This decreased food consumption was measured and, on the average, these dogs consumed 73% of the quantity of food consumed by the control group. The corresponding failure to gain weight in the dogs on this level was proportional to the decrease in food consumption in comparison with the controls. Indirectly some organ to body weight ratios were affected in the dogs an this level. However, there was no difference in organ weights when considered on the absolute basis in grams, and the variation in the ratios may be attributed to the decrease in body weight when compared with the controls. There was an increase in the serum alkaline phosphatase values in the dogs which received 1% Benax 2A1 in their diet. However, there were no accompanying pathological changes in the liver nor any significant difference in the liver/body weight ratio when compared with the controls. Based on the above findings the study NOEL was 131 mg/kg/day.

In the 1976 ss (Klimisch 2), rats were given 0, 50, 100, 200 or 600 mg of powdered analogue DOWFAX XD-8390/kg/day in the diet for 90-91 days. Morphological manifestations which could be associated with ingestion of the test material were observed only at the two higher dose levels, 200 and 600 mg/kg/day and were limited to effects on the kidneys. The weights of the kidneys were significantly increased at the two higher dose levels among the female rats and at the highest dose level among the male rats and were accompanied by some microscopic observations. The kidney morphology and function was not different from that of control rats at the two lower dose levels of the test material. The SGPT activity was increased at all dose levels in the male rats in the absence of any histological or other biochemical evidence of liver damage. Except for these changes no alterations in other parameters were observed at any dose level. Based on these findings the NOAEL in rats was 200 and 600 mg/kg/day for female and male rats, respectively.

In the other 1976 ss (Klimisch 2), dogs were given 0, 50, 100 or 200 mg/kg/day DOWFAX XD 8390 analogue in the diet for 90 days. Effects which could be associated with ingestion of DOWFAX surfactant XD-8390 in the diet for 90 days were limited to the kidneys of male and female dogs at the high dose level, 200 mg/kg/day. Among these dogs, the weight of the kidneys and liver was higher than that of control dogs. The increased weight of these organs was not associated with histological alterations. Thus, no significant toxicological manifestations were observed among any of the groups of dogs at any dose level of DOWFAX surfactant XD-8390. The NOAEL was 200 mg/kg/day for dogs.

DERMAL:

In the 1993 (Klimisch 2) subacute study, the skin irritation test included topical application of 0.5 ml of neat DOWFAX 2A1 to the ear, and to intact and abraded skin on the abdomen of a male New Zealand White rabbit. Five consecutive daily doses of DOWFAX 2A1 were applied to the intact abdominal skin, and the apex of the left pinna, and three consecutive daily applications to the abraded abdominal site. No irritation was observed on the ear. Very slight erythema was observed at the intact site after the last application. Very slight erythema was observed at the abraded site on test day two through test day five. The study was terminated 72 hours after the final dose.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Longest repeat dose study with DOWFAX 2A1 with lowest NOAEL. All durations of exposure to test material showing similar effects in test animals.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
sufficient data available on the registered substance for assessment.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
sufficient data available on the registered substance for assessment.

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

Justification for classification or non-classification

Non-classification based on the chronic NOAEL of 128 mg/kg/day in accordance with the CLP.