Quaternary ammonium compounds, C12-14-alkylethyldimethyl, Et sulfates

Dermal absorption

Administrative data

Endpoint:

dermal absorption in vitro / ex vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From 18 August, 2005 to 17 August, 2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to the OECD guideline 428 as well as in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 428 (Skin Absorption: In Vitro Method)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Quaternary ammonium compounds, benzyl-C12-16-alkyldimethyl, chlorides

EC Number:

270-325-2

EC Name:

Quaternary ammonium compounds, benzyl-C12-16-alkyldimethyl, chlorides

Cas Number:

68424-85-1

IUPAC Name:

N-benzyl-N,N-dimethyltetradecan-1-aminium chloride

Constituent 2

Reference substance name:

Alkyl(C12-C16)dimethylbenzylammonium chloride (i.e., benzyl C12-16 alkyl dimethyl ammonium chloride)

IUPAC Name:

Alkyl(C12-C16)dimethylbenzylammonium chloride (i.e., benzyl C12-16 alkyl dimethyl ammonium chloride)

Details on test material:

- Name of test material: Alkyl(C12-C16)dimethylbenzylammonium chloride (non-radiolabelled ADBAC) and [14C]-ADBAC (radiolabelled ADBAC)
- Analytical purity: non-radiolabelled: 80.5% pure; radiolabelled: 99.7%
- Composition of test material, percentage of components:
- Lot/batch No.: radiolabelled: 990930; non-radiolabelled: J2220459
- Radiochemical purity (if radiolabelling): 99.7%
- Specific activity (if radiolabelling): 25 mCi/mmol; total radioactivity: 1 mCi/mL in etanol
- Storage condition of test material: non-radiolabelled: ambient temperature in dark

Radiolabelling:

yes

Test animals

Species:

human

Details on test animals or test system and environmental conditions:

Human skin membranes, in vitro

Administration / exposure

Type of coverage:

other: Automated flow-through diffusion cell system

Vehicle:

water

Doses:

0.03% (w/w) and 0.3% (w/w), in water

Details on in vitro test system (if applicable):

SKIN PREPARATION
- Source of skin: Eight samples from Plastic Surgery unit, St. Johns Hospital, West Lothian NHS Trust, Livingston, UK
- Ethical approval if human skin: Yes
- Type of skin: Full-thickness human skin (1 upper arm, 2 abdomen and 5 breast)
- Preparative technique: Skin was transferred to Charles River Laboratories on ice and cleaned of subcutaneous fat and connective tissue using scalpel. Skin was washed in cold running tap water and dried using tissue paper. Each sample was then cut into smaller pieces, wrapped in aluminium foil, put into self sealing plastic bags and stored at -20°C until required. Split-thickness membranes were prepared by pinning the full thickness skin, stratum corneum uppermost, onto a raised cork board and cutting at a setting equivalent to 200-400 µm depth using a Zimmer electric dermatome.

PRINCIPLES OF ASSAY
- Diffusion cell: Automated flow-through diffusion cell system (Scott/Dick, University of Newcastle-upon-Tyne, UK)
- Receptor fluid: Tissue culture medium containing approximately 5% (w/v) bovine serum albumin, 1% (w/v) streptomycin (approximately 0.1 mg/mL), and penicillin G (approximately 100 units/mL)
- Solubility of test substance in receptor fluid:
- Flow-through system: Automated
- Test temperature: The mean temperature 19°C (SD=0.8°C)
- Humidity: The mean relative humidity 35% (SD=0.6%)

Results and discussion

Total recovery:

Refer to Table-1

Percutaneous absorption

Percutaneous absorption 1

Dose:

0.03%

Parameter:

percentage

Absorption:

0.05 %

Remarks on result:

other: 24 h

Remarks:

96.80% was not absorbed

Percutaneous absorption 2

Dose:

0.3%

Parameter:

percentage

Absorption:

0.03 %

Remarks on result:

other: 24 h

Remarks:

94.68% was not absorbed

Any other information on results incl. tables

Low dose (0.030%): The mean mass balance was 99.03% of the applied dose (3.09 μg equiv./cm²). The mean dislodgeable dose was 60.53% of the applied dose (1.89 μg equiv./cm²). The mean total unabsorbed dose was 96.80% of the applied dose (3.02 μg equiv./cm²). This consisted of the dislodgeable dose, unexposed skin (0.02%) and the radioactivity associated with the stratum corneum (36.25%). The stratum corneum acted as a good barrier to the test substance as the bulk of the radioactivity (30.26%) was recovered in the outermost 5 tape strips (tape strips 1-5). Considerably less radioactivity was recovered with each of the subsequent 3 groups of tape strips (3.20%, 1.82% and 0.97% in tape strips 6-10, 11-15 and 16-20 respectively), suggesting that the test substance would be sloughed off with the skin in the future. The absorbed dose (0.05%, or <0.01 μg equiv./cm²) was the sum of the receptor fluid (0.05%) and the receptor rinse (<0.01%). Dermal delivery (2.22%, or 0.07 μg equiv./cm²) was the sum of the absorbed dose and the exposed skin (2.18%). There was no apparent lag time and the fluxes ranges from 0.03 to 0.12 ng equiv./cm²/h over the 1 to 24-h exposure period.

 

High dose (0.300%): The mean mass balance was 96.84% of the applied dose (29.91 μg equiv./cm²). The dislodgeable dose was 77.87% of the applied dose (24.05 μg equiv./cm²). The mean total unabsorbed dose was 94.68% of the applied dose (29.24 μg equiv./cm²). This consisted of the dislodgeable dose, unexposed skin (0.17%) and the radioactivity associated with the stratum corneum (16.64%). The stratum corneum acted as a good barrier to the test item as the bulk of the radioactivity (10.86%) was recovered in the outermost 5 tape strips (tape strips 1-5). Considerably less radioactivity was recovered with each of the subsequent 3 groups of tape strips (3.11%, 1.71% and 0.96% in tape strips 6-10, 11-15 and 16-20, respectively), again suggesting that the test item would be sloughed off with the skin in the future. The absorbed dose (0.03%, or 0.01 μg equiv./cm²) was the sum of the receptor fluid (0.03%) and the receptor rinse (<0.01%). Dermal delivery (2.16%, or 0.67 μg equiv./cm²) was the sum of the absorbed dose and the exposed skin (2.13%). There was no apparent lag time. There was no apparent lag time and the fluxes ranges from 0.22 to 0.74 ng equiv./cm²/h over the 1 to 24-h exposure period.

Table-1. Summary of recoveries after 24 h

Test Preparation	Low Concentration	High Concentration
Target test substance concentration (%, w/w)	0.03	0.30
Test substance concentration by Radioactivity (%, w/w)	0.031	0.306
Test preparation application rate (mg/cm^2)^a	10.01	10.09
Test substance application Rate (μg equiv./cm^2)	3.12	30.87
Dislodgeable Dose (% Applied Dose)	60.53	77.87
Unabsorbed Dose (% Applied Dose)	96.80	94.68
Absorbed Dose (% Applied Dose)	0.05	0.03
Dermal Delivery (% Applied Dose)	2.22	2.16
Mass Balance (% Applied Dose)	99.03	96.84
Dislodgeable Dose (μg equiv./cm^2)	1.89	24.05
Unabsorbed Dose (μg equiv./cm^2)	3.02	29.24
Absorbed Dose (μg equiv./cm^2)	<0.01	0.01
Dermal Delivery (μg equiv./cm^2)	0.07	0.67
Mass Balance (μg equiv./cm^2)	3.09	29.91

^a mg of test preparation per cm of skin

 

Applicant's summary and conclusion

Conclusions:

Under the study conditions, following topical application of [14C]-radiolabelled test substance in low (0.03%, w/w) and high (0.3%, w/w) concentration, the mean absorbed dose and mean dermal delivery of [14C]- radiolabelled test substance were 0.05% (<0.01 μg equiv./cm2) and 2.22% (0.07 μg equiv./cm2) of the applied dose for the low concentration test preparation, respectively, and 0.03% (0.01 μg equiv./cm2) and 2.16% (0.67 μg equiv./cm2) of the applied dose for the high concentration test preparation, respectively. The maximum fluxes for the low and high doses were 0.12 ng equiv./cm2/h and 0.74 ng equiv./cm2/h, respectively, at 2 h.

Executive summary:

A guideline compliant in vitro study was conducted to determine the dermal absorption of C12-16 ADBAC. Split-thickness human skin membranes were mounted into flow-through diffusion cells. Receptor fluid was pumped underneath the skin at a flow rate of 1.5 mL/hour. The skin surface temperature was maintained at approximately 32°C. A barrier integrity test using tritiated water was performed and any skin sample exhibiting a permeability coefficient (kp) greater than 2.5 x 10 ^-3 cm/hour was excluded from subsequent absorption measurements. Two test preparations containing [¹⁴C] - radiolabelled test substance (i.e., 0.03% and 0.3%), were applied at an application rate of 10 mg/cm². Absorption was assessed by collecting receptor fluid in hourly intervals from 0-6 hours post dose and then in 2-hourly intervals from 6-24 hours post dose. At 24 hours post dose, exposure was terminated by washing and drying the skin. The stratum corneum was then removed from the skin by 20 successive tape strips. All samples were analysed by liquid scintillation counting. Following topical application of ¹⁴C- radiolabelled test substance in low (0.03%, w/w) and high (0.3%, w/w) concentration test preparations to human skin in vitro, the mean absorbed dose and mean dermal deliveries were 0.05% (<0.01 ηg equivalent/cm²) and 2.22% (0.07 ηg equivalent/cm²) of the applied dose for the low concentration test preparation, respectively, and 0.03% (0.01 ηg equivalent /cm²) and 2.16% (0.67 ηg equivalent/cm²) of the applied dose for the high concentration test preparation, respectively. The stratum corneum acted as a barrier to absorption, with the mean total unabsorbed doses (recovered in skin wash, tissue swabs, pipette tips, cell wash, stratum corneum and unexposed skin) of 96.80 and 94.68% of the applied dose for the low and high concentration test preparations, respectively. The maximum fluxes for the low and high doses were 0.12 ηg equivalent/cm²/hour and 0.74 ηg equivalent/cm²/hour, respectively, at 2 hours.