Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 July 2010 - 30 August 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
Species Mouse, CBA/J strain, inbred, SPF-Quality.
Recognized by the international guidelines as the recommended test system (e.g. OECD, EC, EPA).
Source: Charles River France, L’Arbresle Cedex, France.
Number of animals 20 females (nulliparous and non-pregnant), five females per group.
Age and bodyweight Young adult animals (approx. 10 weeks old) were selected. Body weight variation was within +/- 20% of the sex mean.

Conditions
Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0ºC, a relative humidity of 40-70% and 12 hours artificial fluorescent light and 12 hours darkness per day.
Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70% for relative humidity. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.

Accommodation
Individual housing in labeled Macrolon cages (MI type; height 12.5 cm) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France). Paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) was supplied as cage-enrichment.
The paper was removed on Day 1 prior to dosing and was supplied again after scoring of the ears on Day 3.

Acclimatization period
The acclimatization period was at least 5 days before the start of treatment under laboratory conditions. Accommodation was as described above except that the animals were group housed in Macrolon cages (MIII type; height 18 cm).

Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Water
Free access to tap water.

Results of analysis for diet (nutrients and contaminants), sawdust, paper and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

Study design: in vivo (LLNA)

Vehicle:
other: watery L92
Concentration:
0, 25, 50%
No. of animals per dose:
5
Positive control substance(s):
other: 25% formalin, containing 37% formaldehyde (w/w)

Results and discussion

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
The SI values calculated for the substance concentrations 10, 25 and 50% were 0.7, 0.6 and 1.6 respectively. Since there was no indication that the test substance elicits an SI ≥ 3 when tested up to 50%, Complexation products of sodium tartrate with iron trichloride was considered not to be a skin sensitizer. It was established that the EC3 value (the estimated test substance concentration that will give a SI =3) (if any) exceeds 50%. The positive control group added to the study showed that the vehicle is suitable for eliciting an SI>3 in this batch of animals and with the procedures used for this study.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50% were 272, 240 and 598 DPM respectively. The mean DPM/animal value for the vehicle control group was 384 DPM and a mean DPM/animal value of 4643 DPM was obtained for the positive control group.

Any other information on results incl. tables

Skin reactions / Irritation No irritation of the ears was observed in any of the control animals and animals treated with the test substance. The slight irritation of the ears in the positive control group was considered not to have a toxicologically significant effect on the activity of the nodes. No oedema was observed in any of the animals examined. Macroscopy of the auricular lymph nodes and surrounding area All auricular lymph nodes of the animals treated with the test substance and control animals were considered normal in size. The auricular lymph nodes of the positive control group animals appeared larger in size as compared to the other treated groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals. Body weights Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. The slight body weight loss, noted in several animals, was considered not toxicologically significant. Toxicity and mortality No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The SI values calculated for the substance concentrations 10, 25 and 50% were 0.7, 0.6 and 1.6 respectively.
Since there was no indication that the test substance elicits an SI ≥ 3 when tested up to 50%, Complexation products of sodium tartrate with iron trichloride was considered not to be a skin sensitizer. It was established that the EC3 value (the estimated test substance concentration
that will give a SI =3) (if any) exceeds 50%. The positive control group added to the study showed that the vehicle is suitable for eliciting an SI>3 in this batch of animals and with the procedures used for this study. The six-month reliability check with Alpha-hexylcinnamicaldehyde indicates that the Local Lymph Node Assay as performed at NOTOX is an appropriate model for testing for contact hypersensitivity.
Based on these results Complexation products of sodium tartrate with iron trichloride would not be regarded as a skin sensitizer according to the recommendations made in the test guidelines.
Executive summary:

Assessment of Contact Hypersensitivity to Complexation products of sodium tartrate with iron trichloride in the Mouse (Local Lymph Node Assay).

The study was carried out based on the guidelines described in:

OECD, Section 4, Health Effects, No.429 (2002),

EC, No 440/2008; B42: "Skin Sensitization: Local Lymph Node Assay"

EPA, OPPTS 870.2600 (2003) “Skin Sensitization”.

Test substance concentrations selected for the main study were based on the results of a preliminary study.

In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 10, 25 or 50% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (1% L92)

and another group of five animals received a positive control substance (25% formalin in 1% L92).

Three days after the last exposure, all animals were injected with 3H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and pooled for each animal. After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index (SI) was subsequently calculated for each group.

No irritation of the ears was observed in any of the control animals and animals treated with the test substance. The slight irritation of the ears in the positive control group was considered not to have a toxicologically significant effect on the activity of the nodes. No oedema was observed

in any of the animals examined.

All auricular lymph nodes of the animals treated with the test substance and control animals were considered normal in size. The auricular lymph nodes of the positive control group animals appeared larger in size as compared to the other treated groups.

No macroscopic abnormalities of the surrounding area were noted in any of the animals. Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50% were 272, 240 and 598 DPM respectively. The mean DPM/animal value for the vehicle control group was 384 DPM and a mean DPM/animal value of 4643 DPM was obtained for the positive control group. The SI values calculated for the substance concentrations 10, 25 and 50% were 0.7, 0.6 and 1.6 respectively.

Since there was no indication that the test substance elicits an SI ≥ 3 when tested up to 50%, Complexation products of sodium tartrate with iron trichloride was considered not to be a skin sensitizer. It was established that the EC3 value (the estimated test substance concentration that will give a SI =3) (if any) exceeds 50%. The positive control group added to the study showed that the vehicle is suitable for eliciting an SI>3 in this batch of animals and with the procedures used for this study. The six-month reliability check with Alpha-hexylcinnamicaldehyde indicates that the Local Lymph Node Assay as performed at NOTOX is an appropriate model for testing for contact hypersensitivity.

Based on these results Complexation products of sodium tartrate with iron trichloride would not be regarded as a skin sensitizer according to the recommendations made in the test guidelines.