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Toxicological information

Dermal absorption

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Administrative data

Endpoint:
dermal absorption in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 428 (Skin Absorption: In Vitro Method)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Di-octyl ether
- Physical state: liquid
- Analytical purity: 99.1%
- Lot/batch No.: CE72530027
Radiolabelling:
no

Test animals

Species:
other: in vitro

Administration / exposure

Type of coverage:
open
Vehicle:
unchanged (no vehicle)
Duration of exposure:
24 hours
Doses:
30 µL
Details on study design:
The aim of the present study was to investigate the in vitro penetration of dioctyl ether (DOE) from the product Cetiol OE at human skin of 3 different donors in vitro according to SCCP requirements and to OECD Guideline 428. The test item was only checked for penetration characteristics into the skin, not for permeation properties across the skin into an aqueous acceptor medium. Because of its low water solubility it is not possible to detect the emollient in the usually used acceptor buffer KRB, pH 7.4.
The product provided by the customer was used undiluted. The first step of the study was to establish an analytical method for quantification of DOE. The described analytical method was developed to quantify the compound in the used extraction medium. The method was validated under GLP conditions for this purpose. At the same time the stability of DOE was tested over 24 hours at 25 °C in the extraction medium employed in the penetration studies (methanol). At the beginning of the study, the DOE content in the test item was determined by LC-MS in triplicate. Additionally the content of DOE in Cetiol OE after skin contact was determined. The penetration of DOE from the test item was investigated at human skin from 3 donors in two fold (n=2) for each donor (totalizing n=6) conformable to the SOP M 019 at Across Barriers. The skin specimen was internally encoded according to the corresponding SOP A 005 at Across Barriers. Full thickness skin was used. The skin thickness was measured immediately before
performing the studies. At the end of the penetration experiment the remaining DOE content at the skin surface was determined. For that goal the test item was collected with cotton swabs and transferred to a falcon tube with the extraction medium; this is the so-called wash procedure. After removing residual emollient, the concentration of DOE in the skin - in the Stratum corneum and deeper skin layers - was quantified. The upper corneous layer of the skin was stripped off and the residual skin was cryo-sectioned. The content of DOE in the filter placed under the skin was measured. After the in vitro study a mass recovery was carried out to determine the mass balance and local distribution of DOE in the different skin compartments. For that goal a quotient of total mass of DOE at the end of the study on the skin surface, in Stratum corneum, Epidermis/Dermis and the
used filter versus the applied amount of DOE in the test item at the start of the study was calculated. Parallel to the in vitro studies with DOE at human skin, the permeability of Caffeine was carried out (MEA: multiple endpoint analysis) at a concentration of 10 mg/L in Krebs-Ringer-buffer (KRB) at pH 7.4 (n=2 for 2 skin donors, n=3 for 1 skin donor). Caffeine is a recommended marker molecule of the OECD Guideline for the quality control of human skin. The results of the Caffeine permeation were compared with the permeability coefficients of previous studies on historical human skin membranes of different origin at Across Barriers.
Details on in vitro test system (if applicable):
SKIN PREPARATION
- Source of skin: human skin
- Type of skin: abdomen

PRINCIPLES OF ASSAY
- Diffusion cell: Franz cell
- Test temperature: 25%
- Occlusion: no
- Reference substance(s): caffeine

Results and discussion

Signs and symptoms of toxicity:
not examined
Dermal irritation:
not examined
Absorption in different matrices:
The mass recovery calculations present amounts and percentages of compound which penetrated and remained in the donor compartment. The mean amount of DOE removed from the skin surface (skin wash) ranged from 103.90 % to 120.51 % of the dose applied. The mean recovery in the two first tape strips was 0.20 % ±0.09 % during all performed experiments. In the further 18 tape strips a mean recovery of 0.52 % ±0.27 % was documented. The mean absorbed dose of DOE, sum of the amounts found in the viable epidermis, dermis and filter, was 0.30 % ±0.15 %. That means, most of the test item remains at the skin surface, a small amount migrates into the Stratum corneum and an even smaller amount enters the deeper skin layers.
Total recovery:
The results from the extraction method have shown that methanol was suitable for performing the penetration experiments presenting recovery values which has ranged between 95.01 % to 118.42 % for the higher concentration of DOE, which complies to the acceptance limit 100 ±20 %. For the lower concentration the mean recovery values were between 90.05 % - 125.34 %.
Percutaneous absorption
Dose:
30 µL
Parameter:
percentage
Absorption:
> 0 - < 1 %
Remarks on result:
other: 24 hours

Applicant's summary and conclusion

Conclusions:
Most of the test item remains at the skin surface, a small amount migrates into the Stratum corneum and an even smaller amount enters the deeper skin layers.
Executive summary:

The LC-MS method was successfully developed and validated with regard to method selectivity, linearity, accuracy and precision in the extraction medium employed for the penetration experiments (methanol). Furthermore, the test compound has demonstrated good stability over the period of 24 hours at RT in methanol. The content of DOE was determined in triplicate in the test solution and mean value was 108.0 %, which complies to the acceptance criteria of 100±10 %. The results from the extraction method have shown that methanol was suitable for performing the penetration experiments presenting recovery values which has ranged between 95.01 % to 118.42 % for the higher concentration of DOE, which complies to the acceptance limit 100±20 %. For the lower concentration the mean recovery values were between 90.05 % - 125.34 %.

The mass recovery calculations present amounts and percentages of compound which penetrated and remained in the donor compartment. The mean amount of DOE removed from the skin surface (skin wash) ranged from 103.90 % to 120.51 % of the dose applied. The mean recovery in the two first tape strips was 0.20 %±0.09 % during all performed experiments. In the further 18 tape strips a mean recovery of 0.52 %±0.27 % was documented. The mean absorbed dose of DOE, sum of the amounts found in the viable epidermis, dermis and filter, was 0.30 %±0.15 %. That means, most of the test item remains at the skin surface, a small amount migrates into the Stratum corneum and an even smaller amount enters the deeper skin layers.