Registration Dossier

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:
30 May 2006 - 01 September 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed accrding to guideline and under GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 428 (Skin Absorption: In Vitro Method)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Non-radiolabeled
Test material name: Halamid
Other name: Chloramine T trihydrate
Batch number: 50210
Appearance: white powder
Chemical purity: 99.7%
Supplier: Sigma-Aldrich
Storage conditions: ambient temperature

radiolabeled
Test material name: [14C]-Halamid
Other name: [ring-U-14C]-Chloramine T (trihydrate)
Batch number: CFQ14688
Specific activity: 23 mCi.mmol
Radiochemical purity: 97.2% (HPLC, on 29 March 2006)
Supplier: GE Healthcare
Storage conditions: ≤ 18 ºC
Radiolabelling:
yes

Test animals

Species:
human

Administration / exposure

Type of coverage:
open
Vehicle:
water
Duration of exposure:
8 hours
Doses:
300 or 50 µg/cm2, 3% or 0.5% solutions in water.
No. of animals per group:
6 skin membranes were used per dose and 2 skin membranes were prepared from each donor for each test group (A and B)
Control animals:
no
Details on in vitro test system (if applicable):
SKIN PREPARATION
- Source of skin: Human, 3 donors, freshly excised skin following abdominal surgery.
- Preparative technique: dermatomed to approx. 400μm.
Recorded: - Thickness of skin (in mm): - Membrane integrity
PRINCIPLES OF ASSAY
- Diffusion cell: The skin membranes were placed in 9 mm flow-through automated diffusion cells
- Solubility of test substance in receptor fluid sufficient: exceeds the required solubility by a factor 6.
- Flow-through system: The receptor fluid was pumped at a speed of cu 1.6 mL/h
- Test temperature: The mean skin surface tempenture was 32 ± 1 ºC
- Reference substance(s): none

Results and discussion

Absorption in different matrices:
see: remarks on results including tables and figures
Total recovery:
- Total recovery: The mean recovery was 95.4% (high dose) and 94.0% (low dose)
- Recovery of applied dose acceptable: yes
- Results adjusted for incomplete recovery of the applied dose: no
- Limit of detection (LOD):
- Quantification of values below LOD or LOQ:

Any other information on results incl. tables

Stability:

In the receptor fluid, a rapid degradation of Chloramine-T trihydrate towards p-TSA was observed. A few minutes after preparing a 30mg/l solution 71.7% Chloramine-T trihydrate was left. The concentration further decreased from 48.9%, 29.2% to 14.5% after 7, 24 and 48 hours.

Directly after dosing, the dose solutions were analyzed by radio-HPLC. The relative amount of Chloramine-T trihydrate in the both dose solutions was 94 70 based on W detection. Based on radio-activity, this percentage was lower (ca  72%) due to additional peaks in the beginning of the radio-chromatogram  most probable related to a breakdown of the radio-label. Degradation will most likely take place to p-TSA (as is observed in the stability test using receptor fluid). It is not expected that only the radio-activity associated with the unknown peaks will be absorbed through the skin. A rapid a degradation of Chloramine-T trihydrate to p-TSA on the skin surface was confirmed by analyzing the cotton swab extracts. After 8 hours contact time, only ca 23 and ca 2.3 % Chloramine-T trihydrate was left in the coton swab extracts of the high and low dose group, respectively. It is therefore reasonable to assume that the main compound reaching the receptor fluid will be p-TSA. In the skin wash solution, Chloramine-T trihydrate was relatively stable deceted in time at 92% until 48 hours.

* The exact concetration is described in the method section.

**Total absorption is defined as the amount in the receptor fluid, the receptor compartment wash and the skin membrane, excluding the amount in the tape strips.

The in vitro percutaneous penetration of Halamid

 Halamid  A        B
 concentration measured (g/L)  30.0*  5.0*
 Dose (µg/cm2)  300*  50 * 
 n  6     6   
 penetration into the receptor fluid after 24 h  % of dose  µg/cm2  % of dose  µg/cm2
   4.01  12.0  11.49  5.74
 maximal flux (µg/cm2/h)  0.652     0.367   
 Lag time (h)  2.7     4.0   
 Total absorption (%)**  9.7     20.0   

Applicant's summary and conclusion

Conclusions:
The mean total adsorption of a 3% aqueous Chloramine-T trihydrate solution on human skin is 9.7%. The mean total adsorption of a 0.5% aqueous Chloramine-T trihydrate solution on human skin is 20.0%
Executive summary:

According to OECD 428 and under GLP the percutaneous absorption of  [14C]-Chloramine-T trihydrate in a 3% and 0.5% aqueous solution was evaluated on 6 freshly excised human membranes (2 from one donor). The chemical stability of Chloramine-T trihydrate was determined prior to the conduct of the study aiming at a later analysis of receptor fluid and skin wash fractions obtained in the main study. Skin membranes from three different human donors were used. The exposure time was 8 hours, after which the test compound was washed from the skin and the post-exposure time was 16 hours. Samples were taken from receptor fluid samples, skin wash. Receptor compartment wash, donor compartment wash, tape strips, and digested skin. All collected samples were analyzed with liquid scintillation counting (LSC).To determine the extent of (metabolic) degradation of Chloramine-T trihydrate into p-TSA, samples of the skin wash were analyzed by radio- LC. The total absorption is defined as the amount in the receptor fluid, the receptor compartment wash and the skin membrance, excluding the amount in the tape strips.

A rapid degradation of Chloramine-T trihydrate towards p-TSA was observed in the receptor fluid. Based on UV detection, the relative amount of Chloramine-T trihydrate decreased from 71.7 % established a few, minutes after mixing (t=0) to 14.5 % after 48 hours. In the skin wash solution, Chloramine-T trihydrate was relatively stable detected in time at 92% until 48 hours. For the high dose, the mean penetration of test compound-related radioactivity into the receptor fluid after 24 hours was 12.0 μg/cm2 which was 4.01% of the dose applied. The mean maximal flux was 0.652 μg/cm2/h  the lag time was 2.7 h. The mean total absorption, was 9.7%. For the low dose, the mean penetration of test compound-related radioactivity into the receptor fluid after 24 hours was 5.74 μg/cm2 which was 11.49 % of the dose applied The mean maximal flux was 0.367 μg/cm2/h and the lag time was 4.0 h. Slight differences between donors were observed. The mean total absorption was 20.0 %. The mean recovery was 95.4% (high dose) and 94.0 % (low dose). The mean total adsorption of a 3% aqueous Chloramine-T trihydrate solution on human skin  is 9.7%. The mean total adsorption of a 0.5% aqueous Chloramine T trihydrate solution on human skin is 20.0%