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Toxicological information

Repeated dose toxicity: dermal

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Administrative data

Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 March 1995 to 13 December 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPP 82-2 (Repeated Dose Dermal Toxicity -21/28 Days)
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Physical state: solid
- Storage condition of test material: in the dark at ambient room temperature

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Froxfield Farms (UK) Limited,
- Age at study initiation: 13 to 15 weeks
- Housing: individually in metal cages
- Diet: SDS Rabbit Diet SQC ad libitum
- Water: ad libitum
- Acclimation period: 21 days

ENVIRONMENTAL CONDITIONS
- Temperature: 16.5-22 °C
- Humidity: 40-68 %
- Air changes: approximately 19 per hours
- Photoperiod: 12 hours light/12 hours dark

IN-LIFE DATES: From: 15th March 1995 To: 20th June 1995

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: 12 x 8 cm
- % coverage: approximately 10%
- Type of wrap if used: impervious bandage consisting of gauze covered with 'Elastoplast' elastic adhesive dressing backed with impervious 'Sleek' plaster.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 6 hours on each day
Duration of treatment / exposure:
6 hours per day
Frequency of treatment:
Daily for 21 consecutive days
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Five
Control animals:
yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: three times daily

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: daily

BODY WEIGHT: Yes
- Time schedule for examinations: prior to dosing on day 1 and then weekly intervals

FOOD CONSUMPTION:
- Food consumption for each animal determined: Yes

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to termination
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, overnight
- How many animals: all
- Parameters checked: packed cell volume, haemoglobin, red blood cell count, mean corpuscular haemoglobin concentration, mean corpuscular volume, platelet counts, total white blood cell count, thrombotest, differential white blood cell count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: prior to termination
- Animals fasted: Yes, overnight
- How many animals: all
- Parameters checked: glucose, total protein, albumin, globulin, albumin/globulin ratio, urea nitrogen, creatinine, alkaline phosphatase, glutamic-pyruvic transaminase, gutamic-oxalaxetic transaminase, total bilirubin, sodium, potassium calcium, chloride, inorganic phosphorus, cholesterol

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- The following organs were weighed: adrenals, brain, kidneys, liver, ovaries, spleen, testes.

HISTOPATHOLOGY: Yes
- The appearance of the following tissues was recorded: adrenals, aorta, brain, caecum, colon, duodenum, eyes, gall bladder, heart, ileum, jejunum, kidneys, larynx, liver, lungs, cervical and mesenteric lymph nodes, mammary glands, oesophagus, ovaries, pancreas, pharynx, pituitary, prostate, salivary gland, sciatic nerve, skeletal muscle, skin, spleen, sternum, stomach, testes and epididymides, thymus, thyroid/parathyroid, tongue, trachea, urinary bladder, uterus, vagina and any macroscopically abnormal tissue.
Statistics:
Where appropriate, the following sequence of statistical tests was used:
- if the data consisted predominantly of one particular value, the proportion of values different from the mode was analysed by Fisher's exact test followed by Mantel's test for a trend in proportions. Otherwise:
- Bartlett's test was applied to test for heterogeneity of variance between treatments.
- If no significant heterogeneity was detected, one way ANOVA was performed followed by Williams' test
- If significant heterogeneity was detected and could not be removed by log transformation, Kruskal-Wallis analysis of ranks was used followed by non-parametric equivalent of Williams' test (Shirley's test).

Covariate of analysis of organ weight data was performed using adjusted organ weights where a correlation between organ weight and body weight was established at the 10% level of significance.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
There were no treatment related clinical signs.
Dermal irritation:
no effects observed
Description (incidence and severity):
There were no dermal reactions that were considered to be treatment related. In two animals from the control group, spots were observed during the last few days of the study.
Mortality:
no mortality observed
Description (incidence):
There were no treatment related mortalities.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
There were no statistically significant differences from the control group noted for body weight gains in any of the treatment groups.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
The food consumption for low and high dose group females was slightly higher than control; these differences were however not considered to be 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
Description (incidence and severity):
There were no differences from the controls in the haematological parameters measured that were related to treatment.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Group mean values of significantly different biochemistry values were recorded. Cholesterol levels were higher than control for females of the high dose group. However, individual values were within the expected background range and in the absence of any related biochemical or pathological changes in the liver, this finding was not considered to be of toxicological importance.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no differences from controls in organ weights that were considered to be related to treatment.
Gross pathological findings:
no effects observed
Description (incidence and severity):
The macroscopic examination performed at termination revealed no changes attributable to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
All of the lesions observed by microscopic pathology were minor in nature and not considered to be related to treatment.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined

Effect levels

open allclose all
Key result
Dose descriptor:
NOEL
Effect level:
100 other: mg/kg/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Elevated cholesterol levels in high dose females (considered to be equivocal)
Key result
Dose descriptor:
NOAEL
Effect level:
300 other: mg/kg/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Elevated cholesterol levels in high dose females (considered to be equivocal)

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1: Group mean values body weights (g)

     Dose group (mg/kg/day)
 Time (weeks)  Sex  Control  100  300  1000
 0  M  2823  2929  2954  3029
   F  3144  3206  3083  3189
 1  M  2911  3067  3102  3146
   F  3212  3319  3186  3370
 2  M  3060  3230  3261  3326
   F  3363  3474  3352  3564
 3  M  3200  3341  3337  3489
   F  3475  3631  3485  3631
 Weight gain  M  377  412  383  460
   F  331  425  402  442

Table 2: Group mean values of food consumption (g/animal/week)

     Dose group (mg/kg/day)
 Time (weeks)  Sex  Control  100  300  1000
 1  M  1289  1339  1257  1237
   F  1240  1358  1208  1409
 2  M  1277  1364  1357  1355
   F  1228  1416  1376  1397
 3  M  1210  1289  1217  1262
   F  1258  1357  1321  1313
 Cumulative value  M  3776  3992  3931  3855
   F  3725  4131  3905  4119

Table 3: Significantly different biochemistry values

     Dose group (mg/kg/day)
 Parameter  Sex  Control  100  300  1000
 Glucose (mg/dL)  M  128  139  140  147*
 Protein (g/dL), total  F  5.7  5.9  5.8  6.2*
 Glob  F  2.2  2.4  2.3  2.5*
 Creatinine (mg/dL)  F  1.3  1.2  1.3  1.0*
 GPT (mU/mL)  M  42  34  31*  31*
 GOT (mU/mL)  F  10  16***  16***  9
 Cholesterol (mg/dL)  F  43  50  47  64**

*significantly different from control (P<0.05), Williams' test

**significantly different from control (P<0.01), Williams' test

***significantly different from control (P<0.05), Student's t test

Table 4: Significantly different organ weights

     Dose group (mg/kg/day)
 Organ  Sex  Control  100  300  1000
 Adrenals  M  252  195***  237  232
 Brain  F  9.42  8.85***  8.77***  9.18
 Ovaries  F  0.223  0.350  0.267  0.365*

*significantly different from control (P<0.05), Williams' test

***significantly different from control (P<0.05), Student's t test

Applicant's summary and conclusion

Conclusions:
Under the conditions of the test, the test material had no toxic effect at any of the doses tested. The NOAEL was determined to be 1000 mg/kg/day and the NOEL was determined to be 300 mg/kg/day.
Executive summary:

In a GLP compliant repeat dose (dermal) study conducted in line with standardised guidelines OECD 401 and EPA OPP 82-2, the effects of the repeat dose of the test material in rats was determined.

Under the conditions of the test, no toxic effect was seen at any of the doses tested. The NOAEL was determined to be 1000 mg/kg/day and the NOEL was determined to be 300 mg/kg/day.