Benzenesulfonic acid, mono-C16-24-alkyl derivs., calcium salts , overbased including distillates (petroleum), hydrotreated heavy paraffinic C10-C50

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Exposure was initiated November 28, 1987 and terminal sacrifice was December 21, 1987.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

To evaluate the potential dermal effects of OS#68022C when administered dermally to Sprague-Dawley rats over four weeks to the shaved intact dorsal skin.

GLP compliance:

yes

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Sulfonic acids, petroleum, calcium salts

EC Number:

263-093-9

EC Name:

Sulfonic acids, petroleum, calcium salts

IUPAC Name:

263-093-9

Details on test material:

Sponsors Identification FDRL Identuification No. Specific Gravity

OS#65657B #125092 11-10-87 87-0461 0.96
OS#68022C #125092 11-10-87 87-0462 1.22
OS#82288 #125092 11-10-87 87-0463 0.83
OS#82289 #125092 11-10-87 87-0464 0.91
OS#44321Y #125092 11-10-87 (Label 31) 87-0465 1.04
OS#82287 #125092 11-10-87 (Label 31) 87-0466 0.908
OS#54090A #125092 11-10-87 87-0467 0.88
OS#75788 #125092 11-10-87 87-0468 0.86

Storage Conditions: room temperature in the dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

Test Animals- Source: Sixty-six CD male rats were received in good condition from Charles River Breeding Laboratories, Inc., Wilmington, Massachusetts.

- Age at study initiation: Approximately 6 weeks old

- Weight at study initiation: 140 to 172g

- Fasting period before study:Not applicable

- Housing: Individually in stain less - steel wire-mesh cages suspended above cage paper.

- Diet: The animals were allowed free access to food; basal ration, NIH Open Formula 07 Rat and Mouse Diet (certified), (Zeigler Brothers, Inc, Gardners, PA).

- Water - animals were allowed free access to tap water supplied to the test facility and monitored for contaminants at periodic intervals according to FDRL Standard Operating Procedures.

- Acclimation period: 20-day acclimation and pre test period.

ENVIRONMENTAL CONDITIONS

- Temperature Range: Not advised

- Humidity Range : Not advised

- Air changes (per hr): Not advised

- Photoperiod (hr dark / hrs light): 12 hours continuous light and 12 hours darkness

IN-LIFE DATES: The study was initiated on November 23, 1987 (first day of treatment) and the in-life phase completed on December 21, 1987.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on exposure:

The test articles were applied undiluted to the shaved intact dorsal skin of each test animal five days per week for four weeks for a total of 20 applications.

The test articles were applied evenly by gentle inunction over the test site using a syringe and a glass rod.

The test articles were held in place with 2 layers of gauze and non irritating tape. All rats were fitted with collars to prevent ingestion of the test articles during the exposure period.

At the end of the six hour exposure period, the dressings were removed and the test sites wiped with a mineral oil gauze to remove as much unabsorbed test article as possible. These sites were then wiped with a clean, dry gauge t o remove any excess mineral oil.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

Six hour exposure period.

Frequency of treatment:

Once daily for five days per week over a four week treatment period.

Doses / concentrations

Remarks:

Doses / Concentrations:
1000 mg/kg bodyweight
Basis:
nominal per unit body weight

No. of animals per sex per dose:

Five males per test article

Control animals:

other: Although not identified as such, two groups were considered to be control groups.

Details on study design:

- Dose selection rationale:

The selected route of administration was topical, since dermal contact is a likely route of exposure for the general population.

- Rationale for animal assignment (if not random):

Random

- Rationale for selecting satellite groups:

Not applicable

- Post-exposure recovery period in satellite groups:

Not applicable

- Section schedule rationale (if not random):

Random

Positive control:

No

Examinations

Observations and examinations performed and frequency:

- Mortality Check:
All animals were observed for mortality and overt signs of toxicity twice daily at least 5 hours apart.

- Physical Examination: see appended Appendix 2.
All animals received a detailed physical examination weekly.

- Body Weights: see appended Appendices 3 and 4.
Individual body weights were measured on the first day of test article administration and weekly thereafter.

- Food Consumption: see appended Appendix 5.
Individual food consumption was measured weekly beginning with the first day of test article administration.

Dermal Irritation: see appended Appendix 5.
Application sites were examined for dermal irritation once daily throughout the study period.

Dermal irritation was evaluated according to the method of Draize, 1965.

Sacrifice and pathology:

See attached Pathology Report (Appendix 7):

A complete necropsy examination was conducted on all animals sacrificed at study termination.

Necropsies were performed under the supervision of a board certified veterinary pathologist.

Animals were euthanized using CO2 gas.

Gross necropsy included examination of the external surfaces and orifices, the cranial cavity, carcass, the external and cut surfaces of the brain, the thoracic, abdominal and pelvic cavities and their viscera and the cervical tissues and organs. Untreated and treated skin and all gross lesions were removed from each animal and fixed in 10% neutral buffered formalin.

Other examinations:

Microscopic examination of paraffin embedded hematoxylin and eosin stained tissue sections was performed on the treated and untreated skin and gross lesions from all animals. All tissues were examined by S.W. Thompson, D.V.M., M.S., Diplomate A.C.V.P.

Statistics:

Continuous data including body weight, body weight gain and food consumption w ere analyzed using analysis of variance (Snedecor and Cochran,
1967).

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No test article related signs of toxicity were noted throughout the 4-week treatment period.

Three instances of sores and two observations of hair loss all in the neck region were noted. These findings were attributed to the collars worn by the rats during the 6-hour exposure period.

Dermal irritation:

effects observed, non-treatment-related

Description (incidence and severity):

No dermal irritation was noted when test articles OS#65657B, OS#68022C, OS#82288, OS#82289 and OS#75788 were applied at 1000 mg/kg to the skin for five days per week for four weeks.

Pinpoint scabbed areas were noted on the skin of one animal on study on Day 13 when OS#54090A was dosed at 1000 mg/kg. No other signs of dermal irritation were noted for this test article throughout the study period.

Pinpoint scabbed areas were noted on the skin of one animal on study Days 10 through 18 when OS#82287 was dosed at 1000 mg/kg. No other signs of dermal irritation were noted for this test article throughout the study period.

All animals exhibited dermal irritation when dosed with OS#44321P. Well-defined erythema was noted for one animal on study Day 3 and a second animal on study Day 3. The other 3 animals exhibited very slight erythema on study Day 3.

Other findings noted for this group included pinpoint scabbed areas and dry flaking skin. Two animals exhibited dermal irritation for the remainder of the 26 day study period. For animal numbers 0025, 0021 and 0023, no dermal irritation was noted on study days 20, 23 and 24 through day 28 respectively.

Mortality:

no mortality observed

Description (incidence):

No deaths occurred during the study period.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No differences in mean body weight or body weight gain were noted among the eight treated groups throughout the study period.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Mean food consumption, evaluated as grams per animal per week, was comparable among the eight treated groups throughout the study period

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

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Compound-related findings were noted at necropsy for two animals treated with GS#34321Y.
Animals numbered 0022 and 0023 exhibited pinpoint scabbed areas over the site of administration. These were the same rats that exhibited dermal irritation to the end of the study period when observed during the in-life phase.
The other findings noted at necropsy were instances of sores around the neck. These findings correlate with the in-life findings of sores and hair loss for the same animals. This was attributed to the collars worn during the 6-hour dosing period and not considered treatment related.
No other findings were noted at necropsy.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Compound-related microscopic effects were noted in 3 of the 5 rats treated with OS#44321Y. The lesions were observed on the treated skin sites and consisted of minimal (rat numbers 0023 and 0025) to mild (rat number 0022) multifocal eschars. In rat number 0022, mild multifocal hemorrhages in the underlaying dermis also were noted.

Histopathological findings: neoplastic:

not examined

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

All rats exhibited minimal hyperkeratosis of the epithelium accompanied by minimal acanthosis of the epidermis at the site of test article administration. Since this finding occurred in all groups and with essentially no differences in severity, it was considered a response of the skin to repeated shaving and exposure to a foreign material.

Effect levels

Key result

Dose descriptor:

NOEL

Effect level:

> 1 000 other: mg/kg to the skin

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Dermal applications of OS#68022C did not elicit a test article related effect when applied at a dose level of 1000 mg/kg to the skin for five days per week over a four week treatment period

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Dermal applications of OS#68022C did not elicit a test article related effect as determined by weekly observations for physical changes and skin irritation and weekly determinations of body weight and food consumption when applied at a dose level of 1000 mg/kg to the skin for five days per week over a four week treatment period

OS#54090A and OS#82287 both elicited minimal dermal irritation. These effects appeared to be transitory as they occurred in only one animal for one day (OS#54090A) and nine days (OS#82287). No treatment-related effects were noted at necropsy or microscopically.

OS#44321Y produced signs of dermal irritation in all treated animals throughout the majority of the 28 day study period.

These signs included slight to well defined erythema, pinpoint scabbed areas and dry flaking skin.

Microscopic observations of minimal to mild multifocal eschar were noted in two rats and mild multifocal hemorrhages in the underlaying dermis in one rat.

OS#44321Y was irritating when applied at 1000 mg/kg to the skin five days per week for four weeks.

Executive summary:

Dermal applications of OS#68022C did not elicit a test article related effect as determined by daily observations for physical changes and skin irritation and weekly determinations of body weights and food consumption when applied at a dose level of 1000 mg/kg to the skin for five days per week over a four week treatment period. No treatment-related effects were noted at necropsy or microscopically.