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Toxicological information

Acute Toxicity: dermal

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Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
12/1995-03/1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Pre-guideline method, but data are comprehensible and scientifically acceptable. (GLP: Yes) Bronaugh, R.L., R.F. Stewart and E. R. Congdon (1982) Methods for in vitro percutaneous absorption studies II. Animal Models for Human Skin Toxicology and Applied Pharmacology 62, 481-488
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
dermal absorption in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12/1995-03/1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Pre-guideline method, but data are comprehensible and scientifically acceptable. (GLP: Yes)
Qualifier:
no guideline available
Principles of method if other than guideline:
Skin Permeability In vitro Absorption Through Porcine Ear Skin with MPAAU according to:
Bronaugh, R.L., R.F. Stewart and E. R. Congdon (1982) Methods for in vitro percutaneous absorption studies II. Animal Models for Human Skin Toxicology and Applied Pharmacology 62, 481-488
GLP compliance:
yes (incl. certificate)
Radiolabelling:
no
Species:
other: porcine ears
Strain:
not specified
Sex:
not specified
Details on test animals and environmental conditions:
Porcine ears were obtained from a local slaughter-house on the day of slaughter and before the pigs were steam-cleaned. The outer ear region was washed and cleaned with cold water. After carefully shaving, the skin was removed by dissection. The thickness of the dissected skin was 2-3 millimetre. The surface of the skin which was in contact with the test substance during the permeation-assay was 1.13 cm2.

For the determination of the absorption of the test article the skin was mounted in glass diffusion chambers with an area of 1.13 cm2 (area of skin) and a volume of 7 ml. These chambers are subdivided in an upper part (donor chamber) and in a lower part (receptor chamber). A volume of 7 ml physiological saline (0.9 % NaCl-solution) was placed in the receptor chamber of each diffusion cell, followed by the application of 226 pi of the test article dissolved in water (2 mg/ml) to the donor chambers. The top of the donor chamber was covered with parafilm. The diffusion chambers were placed in an incubator at 37° C. Samples (0.5 ml) were drawn from the receptor chamber after 0, 0.5, 1, 2, 4, 6, 8 and 24 hours and analysed. The volume of the fluid in the receptor chamber was kept constant by the addition of 0.5 ml of fresh receptor fluid to the receptor chamber immediately after removal of each sample.
Type of coverage:
other: pig skin inbetween donor chamber and acceptor chamber
Vehicle:
water
Duration of exposure:
Samples (0.5 ml) were drawn from the receptor chamber after 0, 0.5, 1, 2, 4, 6, 8 and 24 hours and analysed.
Doses:
226 µl of the test material dissolved in water (2 mg/ml) were applied to the donor chamber in the first and second experiment.
No. of animals per group:
2 Experiments, each using 4 glass diffusion chambers in parallel.
Control animals:
no
Details on study design:
No check of integrity of the skin preparations.
Acceptor fluid (0.9% NaCl) of sampling interval 0 hours (blank) showed no detector signal within the retention time of the substance.
Signs and symptoms of toxicity:
not examined
Dermal irritation:
not examined
Dose:
2mg ai/ml
Parameter:
percentage
Absorption:
<= 0 %
Remarks on result:
other: 8 hours
Remarks:
The concentration (receptor fluid) did not exceed the lower limit of detection prior to 8 hours after start of experiments.
Dose:
2mg ai/ml
Parameter:
percentage
Absorption:
< 5 %
Remarks on result:
other: 24 hours
Remarks:
In the 24 hours specimens (receptor fluid) the concentration of the permeated test article varied in the range of 0 to 12.7 % of the amount applied and averaged to 4.6 % +/- 4.2 % (n=8).
Conversion factor human vs. animal skin:
In vitro dermal absorption studies are preferably carried out with human skin preparations; pig skin is considered a suitable alternative (EHC 235: Dermal Absorption, page 109). No interspecies conversion of animal data is necessary.
Percutaneous penetration refers to in vitro experiments and represents the amount of topically applied test substance that is found in the receptor fluid - this quantity is taken as systemically available (R.7C p.156). No conversion of in vitro data is necessary.

Since permeation of the test article through porcine skin was very low a graphical evaluation of the analytical data was impossible. The concentration of the permeated test article did not exceed the lower limit of detection prior to 8 hours after start of experiments.

The permeation rates (slope) and permeation constants (Kp) could not be determined since no measurable penetration of the skin occurred prior to 8 hours.

Conclusions:
MPAAU showed very low penetration through the skin of porcine ears. Within 8 hours no MPAAU were detected in the receptor chamber. Within 24 hours less than 5 % (mean value) of the amount of MPAAUN applied to the test system penetrated.
Executive summary:

The test article MPAAU was assessed for its potential to permeate through porcine skin.

The assay was performed in two independent experiments over a time scale of 0 to 24 hours. The test article was tested at the following concentration (a.i):

Experiment I and II: 0.94 mg/ml in water. Samples were drawn from the receptor chambers following a fixed schedule:

Exp. I and II: 0.0; 0.5; 1.0; 2.0; 4.0, 6.0; 8.0 and 24 hours following the application of the test article.

The samples (0.5 ml each) were labelled and transferred to the sponsor's facilities for subsequent analysis.

Specimens of a pig ear skin penetration study were separated by reversed phase HPLC followed by UV-detection at 200 nm. The separated and detected MPAAU a.i. was quantified by an external calibration function. The bottom quantification limit of the practical work amounted to 1.28 µg/mL MPAAU a.i. in aqueous solution. By means of a serial connection of two analytical HPLC-columns penetrated MPAAU a.i. in the specimens of the 24 h sampling interval were quantified. Within 24 hours less than 5 % (mean value) of the amount of FLOVAN CGN applied to the test system penetrated*. Within 8 hours no MPAAU were detected in the receptor chamber.

*) test substance applied to the test system: 212.44 µg MPAAU a.i.; 452.00 µg MPAAU

5 % of the test substance: 10.62 ug MPAAU a.i.; 22.60 ug MPAAU

Discussion: Since permeation of the test article through porcine skin was very low a graphical evaluation of the analytical data was impossible. The concentration of the permeated test article did not exceed the lower limit of detection prior to 8 hours after start of experiments.

In conclusion, it can be stated that MPAAU showed very low penetration through the skin of porcine ears. Within 8 hours no MPAAU were detected in the receptor chamber. Within 24 hours less than 5 % (mean value) of the amount of MPAAUN applied to the test system penetrated.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: pre-guideline method
GLP compliance:
yes (incl. certificate)
Test type:
other: single 24 hours exposure skin penetration test
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: solution
Details on test material:
technical product

Test animals

Species:
pig

Results and discussion

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
The test article MPAAU was assessed for its potential to permeate through porcine skin.
The assay was performed in two independent experiments over a time scale of 0 to 24 hours. The test article was tested at the following concentration (a.i):
Experiment I and II: 0.94 mg/ml in water. Samples were drawn from the receptor chambers following a fixed schedule:
Exp. I and II: 0.0; 0.5; 1.0; 2.0; 4.0, 6.0; 8.0 and 24 hours following the application of the test article.
The samples (0.5 ml each) were labelled and transferred to the sponsor's facilities for subsequent analysis.

Specimens of a pig ear skin penetration study were separated by reversed phase HPLC followed by UV-detection at 200 nm. The separated and detected FLOVAN
CGN a.i. was quantified by an external calibration function. The bottom quantification limit of the practical work amounted to 1.28 µg/mL FLOVAN CGN a.i. in aqueous solution. By means of a serial connection of two analytical HPLC-columns penetrated FLOVAN CGN a.i. in the specimens of the 24 h sampling interval were quantified. Within 24 hours less than 5 % (mean value) of the amount of FLOVAN CGN applied to the test system penetrated*. Within 8 hours no MPAAU were detected in the receptor chamber.

*) test substance applied to the test system: 212.44 µg MPAAU a.i.; 452.00 µg MPAAU
5 % of the test substance: 10.62 ug MPAAU a.i.; 22.60 ug MPAAU

Discussion: Since permeation of the test article through porcine skin was very low a graphical evaluation of the analytical data was impossible. The concentration of the permeated test article did not exceed the lower limit of detection prior to 8 hours after start of experiments.

In conclusion, it can be stated that MPAAU showed very low penetration through the skin of porcine ears. Within 8 hours no MPAAU were detected in the receptor chamber. Within 24 hours less than 5 % (mean value) of the amount of MPAAU applied to the test system penetrated.