Phosphonic acid, P-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-, sodium salt (1:1)

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Arrival of the Test Item: 16 June 2011 Date of Final Report: 18 October 2011

Reliability:

1 (reliable without restriction)

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

yes

Remarks:

Yes see the atthaced copy of the study, the deviations do not affect the validity of the study

Principles of method if other than guideline:

OECD Guidelines for Testing of Chemicals, number 429, “Skin Sensitisation: Local Lymph Node Assay” (adopted: July 22, 2010) [3]

Commission Regulation (EC) No. 440/2008, L 142, Annex Part B, 30 May 2008 [4]

EPA Health Effects Test Guidelines, OPPTS 870.2600 “Skin Sensitization” EPA 712-C-03-197, March 2003 [5]

EPA Health Effects Test Guidelines, OPPTS 870.1000 “Acute toxicity testing background”, EPA 712-C-02-189, December 2002 [6]

Ministry of Health and Welfare (MHW, Japan): Japanese Guidelines for Nonclinical Studies of Drugs Manual 1995, Yakuji-Nippo Co. Ltd., 1995 (unofficial translation) [7]

Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Guidelines for Preparation of Study Results, Skin Sensitization Studies, Guideline 2-1-6. Notification 12 NohSan No. 8147, as partly revised in 16-Shouan-9260, on March 2005. English
translation by IAI:ACIS on 17 October 2005 [8]

[1] Chemikaliengesetz (“Chemicals Act”) of the Federal Republic of Germany, Appendix 1 to § 19a as amended and promulgated on June 20, 2002 (BGBI. I Nr. 40 S. 2090), revised 31 October 2006 (BGBI. I Nr. 50 S. 2407).

[2] OECD Principles of Good Laboratory Practice (as revised in 1997); OECD Environmental Health and Safety Publications; Series on Principles of Good Laboratory Practice and Compliance Monitoring - Number 1. Environment Directorate, Organisation for Economic Co-operation and Development, Paris 1998

[3] OECD Guidelines for Testing of Chemicals, number 429, “Skin Sensitisation: Local Lymph Node Assay” (adopted: July 22, 2010)

[4] Commission Regulation (EC) No 440/2008, L 142, Annex Part B of 30 May 2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)

[5] EPA Health Effects Test Guidelines, OPPTS 870.2600 Skin Sensitization, EPA 712-C-03-197, March 2003, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)

[6] EPA Health Effects Test Guidelines, OPPTS 870.1000 Acute Toxicity Testing - Background, EPA 712-C-02-189, December 2002, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)

[7] Ministry of Health and Welfare (MHW, Japan): Japanese Guidelines for Nonclinical Studies of Drugs Manual 1995, Yakuji-Nippo Co. Ltd., 1995 (unofficial translation)

[8) Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Guidelines for Preparation of Study Results, Skin Sensitization Studies, Guideline 2-1-6. Notification 12 NohSan No. 8147, as partly revised in 16-Shouan-9260, on March 2005. English translation by IAI:ACIS on 17 October 2005

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann, 33178 Borchen, Germany
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: No details provided in report
- Housing: The animals were kept in groups of 5 animals in IVC cages, type II L, polysulphone cages on Altromin saw fibre bedding (lot no. 060411)
- Diet (e.g. ad libitum): Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1455)
- Water (e.g. ad libitum): Free access to tap water, sulphur acidified to a pH value of approx. 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals) (Certificates of food, water and bedding are filed at BSL BIOSERVICE)
- Acclimation period: Adequate acclimatisation period (at least five days)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

- The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on Animal Welfare [6] the animals are bred for experimental purposes.
- Full barrier in an air-conditioned room

[6] EPA Health Effects Test Guidelines, OPPTS 870.1000 Acute Toxicity Testing - Background, EPA 712-C-02-189, December 2002, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)

Vehicle:

other: Due to the solubility properties of the test item 2% carboxymethylcellulose (CMC) in aqua ad inject was used as vehicle. (CMC, Alfa Aesar, lot no. 10159638, aqua ad inject, Berlin-Chemie, lot no. 0195A191, expiry date; 04/2013)

Concentration:

Dose Groups: 3 test groups (3 different concentrations), 1 positive control group (25% alpha-hexylcinnamaldehyde in 2% CMC) and 1 negative control group (vehicle) were tested.

No. of animals per dose:

Number of animals: 5 mice / group
3 mice / preliminary test

Details on study design:

Test Regime

Topical Application: Each mouse was treated by topical application of 25 µL of the selected solution to the entire dorsal surface of each ear. Topical applications were performed once daily over three consecutive days.

Administration of 3H-Methyl Thymidine: Five days after the first topical application all mice were dosed with 20 µCi 3H-methyl thymidine by intravenous injection (tail vein) of 250 µL of 3H-methyl thymidine, diluted to a working concentration of 80µCi/mL.

Preparation of Cell Suspension: Approximately 5 hours after the injection of 3H-methyl thymidine all mice were sacrificed by cervical dislocation, The draining “auricular lymph nodes” were excised, individually pooled for each animal (2 lymph nodes per animal) and collected in phosphate buffered saline (PBS). A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through polyamide gauze (200 mesh size). After washing the gauze with PBS the cell suspension was pelleted in a centrifuge. The supernatant was discarded and the pellets were resuspended with PBS. This washing procedure was repeated.

After the final wash each pellet was resuspended in approx. 1 mL 5% TCA at approx. 40 C for approximately 18 hours for precipitation of macromolecules. Each precipitate was once washed again, resuspended in 1 mL 5% TCA and 7 mL scintillation fluid was added. Then this solution was transferred into scintillation vials and stored at room temperature overnight.

Determination of Incorporated 3H-Methyl Thymidine: The 3H-methyl thymidine - incorporation was measured in a B-counter and expressed as the number of disintegrations per minute (DPM). Similarly, background H-methyl thymidine levels were also measured (5% TCA). Determination of radioactivity was performed individually for each animal.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

other: 2% Carboxymethylcellulose (CMC) in aqua ad injectionem served as negative control (see attached report)

Statistics:

N/A

Positive control results:

Alpha hexylcinnamaldehyde was the positive control used in the study. Details of the results are described in the study attached.

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

For individual data see Table 3 in Appendix of attached report. One of the three tested concentrations of the test item corresponded to a stimulation index of 3 and one of the concentrations exceeded the stimulation index of 3. The stimulation index at a concentration of 6.25% was 1.3 The stimulation index at a concentration of 12.5% was 4.1 The stimulation index at a concentration of 25% was 3.0

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: see Remark

Remarks:

For individual data see Table 3 in Appendix of attached report. Mean Values: Negative control: 1206.6 Positive control: 10693.8 FHP-OHS (Concentration: 6.25%): 1568.5 FHP-OHS (Concentration: 12.5%): 4941.6 FHP-OHS (Concentration: 25%): 3546.8 Background Szinti and TCA: 12.2

All animals survived throughout the test period without showing any clinical signs. For individual data see Table 5 in the appendix of the attached report.   Body Weight Development: All animals showed the expected weight development, which includes a weight loss of up to 2 g throughout the study. For individual data see Table 4 in the appendix of the attached report.

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The EC3 value (derived by linear interpolation) could not be calculated.

One of the tested concentrations corresponded to a stimulation index of 3 and one concentration exceeded the stimulation index of 3. Consequently, according to OECD 429 [3] the test item FHP-OHS is considered to be a dermal sensitiser.

Since it was not possible to calculate the EC3 value, the test item cannot be classified into any of the sub-categories (category 1A or B) according to Commission Regulation (EU) No 286/2011 [6] and OECD-GHS (Globally Harmonized Classification System) [12]. Therefore, according to Commission Regulation (EU) No 286/2011 [6] and OECD-GFTS [12], the test item FHP-OHS is classified into Category 1 and has obligatory labelling requirement for skin sensitisation.

For details of the classification criteria see Evaluation of Results.

[3] OECD Guidelines for Testing of Chemicals, number 429, “Skin Sensitisation: Local Lymph Node Assay” (adopted: July 22, 2010)

[6] EPA Health Effects Test Guidelines, OPPTS 870.1000 Acute Toxicity Testing - Background, EPA 712-C-02-189, December 2002, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)

[12] OECD-GHS - Globally Harmonized System of Classification and Labelling of Chemicals. Third revised edition, United Nations. New York / Geneva, July 2009

Executive summary:

On the basis of the test results given below and in conformity with the criteria given in Commission Regulation (EU) No 286/2011 [6] as well as OECD-GHS (Globally Harmonized Classification System) [12] the substance should be: classified into category 1   Based on the results of the preliminary test the test item was assessed for sensitising properties at concentrations of 6.25 %, 12.5 % and 25 % (w/v).   At the daily clinical observation the animals did not show any visible clinical symptoms and no case of mortality was observed.   Species/strain: Mice, CBA/CaOlaHsd Number of animals: 20/main test Vehicle: AOO (4:1 (v/v) acetone/olive oil)   Summary Results: One of the three tested concentrations of the test item reached the stimulation index of 3, one of the concentrations exceeded the stimulation index of 3.   The stimulation index at a concentration of 6.25% was 1.3 The stimulation index at a concentration of 12.5% was 4.1 The stimulation index at a concentration of 25% was 3.0   Conclusion: The EC3 value (derived by linear interpolation) could not be calculated.   One of the tested concentrations corresponded to a stimulation index of 3 and one concentration exceeded the stimulation index of 3. Consequently, according to OECD 429 [3] the test item FHP-OHS is considered to be a dermal sensitiser.   Since it was not possible to calculate the EC3 value, the test item cannot be classified into any of the sub-categories (category 1A or B) according to Commission Regulation (EU) No 286/2011 [6] and OECD-GHS (Globally Harmonized Classification System) [12]. Therefore, according to Commission Regulation (EU) No 286/2011 [6] and OECD-GHS [12], the test item FHP-OHS is classified into Category 1 and has obligatory labelling requirement for skin sensitisation.   For details of the classification criteria see Evaluation of Results.   [3] OECD Guidelines for Testing of Chemicals, number 429, “Skin Sensitisation: Local Lymph Node Assay” (adopted: July 22, 2010)   [6] EPA Health Effects Test Guidelines, OPPTS 870.1000 Acute Toxicity Testing - Background, EPA 712-C-02-189, December 2002, United States, Environmental Protection Agency, Prevention, Pesticides and Toxic Substances (7101)   [12] OECD-GHS - Globally Harmonized System of Classification and Labelling of Chemicals. Third revised edition, United Nations./, July 2009

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

Migrated from Short description of key information:
Evalaution of FHP-OHS to OECD 429 has shown the substance to be a skin sensitizer Cat 1 under CLP/GHS regulation

Justification for selection of skin sensitisation endpoint:
Only study