Registration Dossier
Registration Dossier
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EC number: 950-347-5 | CAS number: -
Endpoint summary
Administrative data
Description of key information
Adequate substance specific data are available for assessing the skin sensitisation potential of Santicizer P1700.
Skin sensitization: Not a dermal sensitizer
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2020-01-02 to 2020-01-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 22nd July 2010
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- Deviations:
- no
- GLP compliance:
- yes
- Type of study:
- mouse local lymph node assay (LLNA)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Sponsor (Valtris Speciality Chemicals, Lot #: 5084; PSL REFERENCE NO.:191121-1H)
- Expiration date of the lot/batch: 2020-09-05
- Purity test date: Not specified
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: stored at room temperature
- Stability under storage conditions: Test substance was stable for the duration of testing
- Stability under test conditions: Test substance was stable for the duration of testing
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: Solubility testing conducted by the laboratory indicated that the test substance was soluble in acetone/olive oil (4:1 v/v) AOO.
FORM AS APPLIED IN THE TEST (if different from that of starting material) : Clear liquid - Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Animals received from Envigo RMS Inc. on December 11, 2019 (Preliminary Irritation Group Animals) and on December 18, 2019 (Test and Control Group Animals)
- Females (if applicable) nulliparous and non-pregnant: Yes. Females ,nulliparous and non-pregnant
- Microbiological status of animals, when known: Not specified
- Age at study initiation: Preliminary Animals: Young adult (12 weeks); Test and Control Animals: Young adult (11 weeks) at experimental start.
- Weight at study initiation: Test and Control Animals: 19.1 - 25.3 grams at experiment start
- Housing: individually housed in plastic solid bottom cages during the dosing and resting phase of the study. After final weighing until sacrifice, animals were housed in their respective dose groups in plastic cages with bedding. Bedding in the plastic, solid bottom cages was changed at least once per week. Enrichment (e.g., nesting material) was placed in each cage. All caging conformed to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals (Natl. Res. Council, 2011).
- Diet (e.g. ad libitum): Envigo Teklad Global 16% Protein Rodent Diet® #2016 ad libitum
- Water (e.g. ad libitum): Filtered tap water supplied ad libitum.
- Acclimation period: 21 or 22 days
- Indication of any skin lesions: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-23ºC
- Humidity (%): 38-51%
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle
- IN-LIFE DATES: From: 2019-12-11 To: 2020-01-14 - Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- Concentrations of 10%, 25%, 50% and 100% were selected for the main test based on results of the preliminary screening test. Dilutions of the test substance were prepared as w/w mixtures in AOO. The vehicle control, AOO, and a single concentration of a 25% w/w mixture of the positive control HCA in AOO were also prepared. All dosage preparations were freshly prepared on the day of application.
- No. of animals per dose:
- Preliminary Irritation: 2/group
Test (4 groups): 5/group
Vehicle (Negative) Control: 5/group
Positive Control: 5/group - Details on study design:
- PRE-SCREEN TESTS:
- Compound solubility: The test substance, as received (neat), was mixed well prior to use. Solubility testing conducted by the laboratory indicated that the test substance was soluble in acetone/olive oil (4:1 v/v) AOO. All preparations were mixed well prior to dosing.
- Irritation: The test substance concentration of 100% was tested to determine the highest achievable level. This concentration did not produce excessive local irritation.
- Systemic toxicity: The test substance concentration of 100% was tested to determine the highest achievable level. This concentration did not produce overt systemic toxicity.
- Ear thickness measurements & erythema scores: The ears of each mouse were evaluated for erythema and edema pre-dose on Day 1 and on Days 2, 3, and 6 according to the modified Draize scoring system (Draize, Woodard, & Calvary,1944).
25 µL of the test substance or the vehicle alone was applied to the dorsum of both ears of each mouse for three consecutive days. Application was done using an appropriate size I micropipette to accurately deliver25 µL. The dose was gently spread as evenly as possible over the dorsal surface of the ear using the tip of the pipette. No treatment was made on Days 4 and 5. On Day 6, the sites for each mouse were evaluated for local reactions (erythema & edema). Animals were observed daily for signs of toxicity. The Study Director used this data in conjunction with any pre-existing data to select the four concentrations to be tested. The test substance at 10%, 25%, and 50% w/w mixtures in AOO and the neat test substance were selected for the main test.
MAIN STUDY
Selection of Animals / Dose Levels
Prior to dosing, the animals were weighed and the ears were checked for any abnormalities or clinical signs of diseases or injury. Thirty healthy naïve female mice without pre-existing ear irritation were selected and distributed (5 mice per group) into the following groups:
Group Purpose Concentration
1 Vehicle Control 0%
2 Positive Control Substance 25% HCA
3 Test Substance 10%
4 Test Substance 25%
5 Test Substance 50%
6 Test Substance 100%
Concentrations were selected based on toxicity, solubility, irritancy, and viscosity
Sample Preparation
Concentrations of 10%, 25%, 50% and 100% were selected for the main test based on results of the preliminary screening test. Dilutions of the test substance were prepared as w/w mixtures in AOO. The vehicle control, AOO, and a single concentration of a 25% w/w mixture of HCA in AOO were also prepared. All dosage preparations were freshly prepared on the day of application.
Test Substance Application
Beginning on Day 1, a quantity of 25 µL of the appropriate test substance concentration, the positive control substance, or the vehicle alone was applied to the dorsum of both ears of each mouse once per day for three consecutive days (Days 1, 2,and3) using a micropipette. During application, the material was gently spread as evenly as possible over the dorsal surface of the ear using the tip of the pipette.
Dermal Scoring
Prior to each application (Days 1, 2,and 3) and on Day 6, the ears were evaluated for erythema and edema according to the modified Draize scoring system (Draize, Woodard, & Calvary,1944).
3H-methyl Thymidine Injections
On Day6 of the study (three days after the final topical application) 250 µL of sterile phosphate buffered saline (PBS) containing 20 µCi of 3H-methylthymidine was injected intravenously via the tail vein of each mouse.
Lymph Node Assessment
Approximately five hours after the injections, all test and control mice were euthanized via overdose of inhaled Isoflurane and the draining auricular lymph nodes were excised. The lymph nodes were evaluated for each individual mouse. A single cell suspension of lymph node cells (LNC) was prepared in PBS by gently massaging the lymph nodes between the frosted ends of two microscope slides over a collection vessel. The slides were then rinsed briefly with PBS into the vessel.
The contents of the vessel were transferred to a centrifuge tube and washed with an excess of PBS and centrifuged for approximately 10 minutes at 1800 rpm, with an relative centrifugal force (RCF) of 489G. This process was carried out twice. In both cases, the supernatant was decanted and discarded following each centrifugation. After the second wash, 5 mL of the 5% trichloroacetic acid (TCA) in distilled water was then added to the sediment and the tube was vortexed briefly. The DNA was then precipitated in the 5% TCA in distilled water at approximately 4°C overnight (approximately 18 hours).
Following the overnight precipitation of the DNA, the tubes were centrifuged again for approximately 10 minutes at 1800 rpm and the supernatant was discarded. The resulting precipitate was re-suspended using 1 mL of the 5% TCA in distilled water and transferred to 10 mL of scintillation fluid. Incorporation of 3H-methyl thymidine was measured by β-scintillation counting and expressed as disintegrations per minute, minus background dpm.
In-life Observations
All test, control and preliminary mice were observed for signs of mortality, gross toxicity, and/or behavioral changes daily. Preliminary mice were euthanized via CO2 inhalation and all test and control mice were euthanized via overdose of inhaled Isoflurane anaesthetic on Day 6.
Body Weights
Individual body weights of test and control animals were recorded on Day 1 (initial) shortly before test substance application and prior to IV injections on test Day 6.
EVALUATION
The mean and standard deviation of the dpm values were calculated for each dose group. A stimulation index (SI) was derived for each experimental group by dividing the mean dpm of each experimental group by the mean dpm of the vehicle control group. Any test substance that produces an SI 3 in the LLNA is normally considered “positive” for dermal sensitization potential (Kimberetal., 1994).
The EC3 value was not calculated since all dose levels induced a stimulation index of less than 3.0. - Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Statistics:
- Statistical analysis was performed on the DPM minus background values. Significance was judged at p < 0.05. The treated groups and negative vehicle control group were compared using a One-Way Analysis of Variance (ANOVA), followed by comparison of the treated groups to control by Dunnett’s t-test for multiple comparisons. Where variances are considered significantly different by Bartlett’s test, groups were compared using a non-parametric method (Kruskal-Wallis non parametric analysis of variance followed by Dunn’s test) (INSTAT Biostatistics, Graph Pad Software, San Diego, CA). Outlier analysis was conducted using Grubbs (1969).
- Positive control results:
- Group 2 (Positive Control – 25% HCA in AOO): Very slight erythema (score of 1) was evident at four positive control sites on Day 2, at all sites on Day 3 and at four sites on Day 6. Slight edema (score of 1) was present at two sites on Day 6. The positive control (HCA) at 25% produced a dermal sensitization response in mice (SI = 5.57). Therefore, the LLNA test system was valid for this study with Santicizer Platinum P-1700.
- Key result
- Parameter:
- SI
- Value:
- < 3
- Test group / Remarks:
- 100%
- Remarks on result:
- other: No skin sensitisation potential
- Key result
- Parameter:
- SI
- Value:
- < 3
- Test group / Remarks:
- 50%
- Remarks on result:
- other: No skin sensitisation potential
- Key result
- Parameter:
- SI
- Value:
- < 3
- Test group / Remarks:
- 25%
- Remarks on result:
- other: No skin sensitisation potential
- Key result
- Parameter:
- SI
- Value:
- < 3
- Test group / Remarks:
- 10%
- Remarks on result:
- other: No skin sensitisation potential
- Key result
- Parameter:
- EC3
- Remarks on result:
- other: EC3 was not calculated since all dose levels induced a stimulation index of <3.0
- Cellular proliferation data / Observations:
- CELLULAR PROLIFERATION DATA
Preliminary Test: Preliminary irritation group scoring results are presented in Table 1.
Main Test:
Group 1 (Vehicle Control—AOO): No dermal irritation was observed for any of the vehicle control sites.
Group 2 (Positive Control—25% HCA in AOO): Very slight erythema (score of 1) was evident at four positive control sites on Day 2, at all sites on Day 3 and at four sites on Day 6. Slight edema (score of 1) was present at two sites on Day 6.
Group 3 (10% Test Substance in AOO): No dermal irritation was observed for any of the test sites.
Group 4 (25% Test Substance in AOO): No dermal irritation was observed for any of the test sites.
Group 5 (50% Test Substance in AOO): No dermal irritation was observed for any of the test sites.
Group 6 (100% Test Substance): No dermal irritation was observed for any of the test sites.
DETAILS ON STIMULATION INDEX CALCULATION
Main Test: Treatment of mice with 10%, 25%, 50% and 100% of Santicizer Platinum P-1700 resulted in stimulation index values of 0.64, 1.14, 1.21, and 1.09, respectively. As a stimulation index (SI) of less than 3.0 was observed in all the treatment groups, the test substance was not considered positive for a dermal sensitization potential. The positive control (HCA) at 25% produced a dermal sensitization response in mice (SI =5.57). Therefore, the LLNA test system was valid for this study with Santicizer Platinum P-1700.
EC3 CALCULATION
The EC3 value was not calculated since all dose levels induced a stimulation index of less than 3.0.
CLINICAL OBSERVATIONS:
Preliminary Test: All preliminary animals appeared active and healthy. Very slight erythema (score of 1) was evident at two test sites on Day 3. Preliminary irritation group scoring results and individual in-life observations are presented in Tables 1 and 2.
Main Test: All test and control animals appeared active and healthy throughout the study.
BODY WEIGHTS
Main test: Five mice from the test groups, two mice from the vehicle control group, and one mouse from the positive control group lost body weight during the study. All other mice gained body weight during the study. Results are presented in Table 3. - Interpretation of results:
- GHS criteria not met
- Conclusions:
- Treatment of mice with 10%, 25%, 50% and 100% of Santicizer Platinum P-1700 resulted in stimulation index values of 0.64, 1.14, 1.21, and 1.09, respectively. As a stimulation index (SI) of less than 3.0 was observed in all the treatment groups, the test substance was not considered positive for a dermal sensitization potential. The positive control (HCA) at 25% produced a dermal sensitization response in mice (SI =5.57). Therefore, the LLNA test system was valid for this study with Santicizer Platinum P-1700. The EC3 value was not calculated since all dose levels induced a stimulation index of less than 3.0.
Based on the results observed, the test material Santicizer Platinum P-1700 was not considered to be a contact dermal sensitizerin the mouse local lymph node assay. - Executive summary:
The dermal sensitization potential of the test material (Santicizer Platinum P-1700) was evaluated in a key OECD Guideline 429 study using the mouse local lymph node
assay (LLNA). Three concentrations of the test substance (10%, 25% and 50% w/w) in acetone/olive oil (4:1 v/v) (AOO), the neat test substance (100%), and the vehicle alone were topically applied to twenty-five healthy test CBA/J female mice (five/group) for three consecutive days. Three days after the last application, the mice were given an IV injection containing 20 µCi of 3H-methyl thymidine. Approximately five hours later, all animals were euthanized via an overdose of inhaled Isoflurane and the draining (auricular) lymph nodes were harvested and prepared for analysis in a scintillation counter. The results are presented in disintegrations per minute per mouse (dpm/mouse). Each animal’s ears were also evaluated for erythema and edema prior to each application and again on Day 6, prior to the IV injection. A positive control group (five animals) was maintained under the same environmental conditions and treated with a 25% w/w mixture of HCA in AOO in the same manner as the test animals.
All preliminary test animals appeared active and healthy. Very slight erythema (score of 1) was evident at two test sites on Day 3. All test and control animals in the main test appeared active and healthy throughout the study. In the main test, five mice from the test groups, two mice from the vehicle control group, and one mouse from the positive control group lost body weight during the study. All other mice gained body weight during the study.
No dermal irritation was observed for any of the vehicle control sites Group 1 (Vehicle Control-AOO). No dermal irritation was observed for any of the test sites in animals in the test material treated groups (10%, 25%, 50%, and 100% w/w). In Group 2 (Positive Control—25% HCA in AOO), very slight erythema (score of 1) was evident at four positive control sites on Day 2, at all sites on Day 3 and at four sites on Day 6. Slight edema (score of 1) was present at two sites on Day 6.
Treatment of mice with 10%, 25%, 50% and 100% of Santicizer Platinum P-1700 resulted in stimulation index values of 0.64, 1.14, 1.21, and 1.09, respectively. As a stimulation index (SI) of less than 3.0 was observed in all the treatment groups, the test substance was not considered positive for a dermal sensitization potential. The positive control (HCA) at 25% produced a dermal sensitization response in mice (SI =5.57). Therefore, the LLNA test system was valid for this study with Santicizer Platinum P-1700. The EC3 value was not calculated since all dose levels induced a stimulation index of less than 3.0.
Based on the results observed, the test material Santicizer Platinum P-1700 was not considered to be a contact dermal sensitizer in the mouse local lymph node assay.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 09/06/2015 - 25/09/2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Deviations:
- no
- Principles of method if other than guideline:
- There were no amendments to the protocol
- GLP compliance:
- yes
- Type of study:
- Buehler test
- Justification for non-LLNA method:
- EPA Guideline OPPTS 870.2600 delayed contact dermal sensitization test in guinea pigs that followed sound scientific principles.
- Specific details on test material used for the study:
- Label Identity: Santicizer P-1700
Batch No: VSC1002-2
Supplied by: Valerus Specialty Chemicals
Data received: 12/05/15
Storage: Room temperature and humidity
Description: Clear light-yellow liquid
Sample preparation: Induction and challange - The test article was used as received. - Species:
- guinea pig
- Strain:
- Hartley
- Remarks:
- Albino
- Sex:
- male/female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Elm Hill Breeding Labs, Inc., Chelmsford, MA
- Age at study initiation:
- Weight at study initiation: Male: 315-374 g - Female: 288-340 g
- Housing: suspended wire cages
- Diet (e.g. ad libitum): PMI Guinea Pig Chow (Diet #5025) - ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): not specified
- Humidity (%): not specified
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hr light and 12 hr dark
IN-LIFE DATES: From: 16/05/15 to 16/07/15 - Route:
- epicutaneous, occlusive
- Vehicle:
- unchanged (no vehicle)
- Concentration / amount:
- 100% / 0.4mL
- Day(s)/duration:
- 6 Hours
- Route:
- epicutaneous, occlusive
- Vehicle:
- unchanged (no vehicle)
- Concentration / amount:
- 100%/ 0.4mL
- Day(s)/duration:
- 6 hr
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- Group 1 - induction and challenge (10 females and 10 males)
Group 2 - naive to induction (5 females and 5 males) - Details on study design:
- Method Synopsis: 15 healthy male and 15 healthy female Hartley Albino Guinea Pigs were assigned to the study. Group 1 (10 males and 10 females) was induced with Santicizer P1700 at a concentration of 100%. Group 2 (5 males and 5 females) was not induced and served as a naïve control. Group 1 received three topical induction applications, one per week for three weeks. Skin reactions of the animals in Group 1 were recorded 24 and 48 hours following patch removal. Based on the results of the induction application, 100% was chosen as the highest non-irritating concentration for the challenge and was administered to both groups two weeks after the third induction. The skin reactions of all animals were recorded at 24 and 48 hours following patch removal. Body weights were recorded pretest and at termination. All animals were observed once per day for mortality, toxicity and systemic observations. The sensitivity of guinea pigs to a positive control, 85% α-hexylcinnemaldehyde (HCA) is confirmed in this laboratory (approximately every six months).
Dosing
Induction Group 1. 100% . Ten males and 10 females in Group 1 were dosed with 0.4ml of the test article. The dose was applied to the left shoulder area (site 1) using a 25mm Hilltop Chamber which is designed to keep the test article on a 25mm area of the site. The chamber contained a cotton pad used to facilitate contact of the liquid test article with the site. The chamber was covered with a strip of rubber dental dam sufficient to cover the treated sites. The torso was wrapped with no-irritating tape to provide occlusion. After 6hr the dam and test article were removed. Any residual test article was cleansed from the sites with distilled water and the sites were dried with soft toweling. This procedure was performed once per week on the same day each week for a three week period, a total of three 6-hour exposures.
Group 2 Five males and five females were intreated for the three week induction period and served as the naïve control (upon challenge dose).
Fourteen days after the last induction exposure, animals in both Groups 1 and 2 were dosed using the same procedure as the induction phase. Based on the results of induction, 100% was chosen as the highest non-irritating concentration for the challenge. The [challenge] doses were applied to a naïve site on the lower left dorsal area (Site 3).
Results were assessed according to GHS skin irration criteria.
- Challenge controls:
- Yes - naive to induction, 5 males and 5 females
- Positive control substance(s):
- yes
- Remarks:
- 85% alpha hexyl-cinnemaldehyde
- Positive control results:
- HCA Incidence index 24 and 48 hr: 0.40; Severity index 24 and 48 hr: 0.48
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 4
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to very faint
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 2
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to very faint. Soiling and yellowing of anogenital area
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 14
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to moderate
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 14
- Total no. in group:
- 20
- Clinical observations:
- Soiling and yellowing of anogenital area
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- other: 3rd reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 12
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to moderate
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- other: 3rd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 14
- Total no. in group:
- 20
- Clinical observations:
- Soiling and yellowing of anogenital area
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 5
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to faint
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 6
- Total no. in group:
- 20
- Clinical observations:
- Erythema was absent to faint. Soiling and yellowing of anogenital area
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- other: Naive challenge
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- Untreated
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Clinical observations:
- Erythema was absent to very faint
- Key result
- Reading:
- other: Naive challenge
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- Untreated
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Clinical observations:
- Erythema was absent to very faint. Soiling and yellowing of anogenital area
- Key result
- Reading:
- other: positive control historic data retested every 6 months
- Hours after challenge:
- 24
- Group:
- positive control
- Dose level:
- 85% HCA
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- incidence index 0.40
- Key result
- Reading:
- other: positive control historic data retested every 6 months
- Hours after challenge:
- 24
- Group:
- positive control
- Dose level:
- 85% HCA
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- Severity index 0.48
- Key result
- Reading:
- other: positive control historic data retested every 6 months
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- 85% HCA
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- Incidence index 0.40
- Key result
- Reading:
- other:
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- 85% HCA
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- Severity index 0.48
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- Based on the results observed, Santicizer P1700 was determined to be a dermal sensitizer in the under the conditions of this delayed contact dermal sensitization test in guinea pigs.
- Executive summary:
In a key EPA OPPTS Guideline 870.2600 delayed contact dermal sensitization study, the potential of the test material (Santicizer P1700) to promote skin sensitization reactions following repeated applications was evaluated in guinea pigs using the Buehler method.
Healthy male and female Hartley Albino guinea pigs were divided into two groups. Group 1 (10 males and 10 females) was induced with the test material at a concentration of 100%. Group 2 (5 males and 5 females) animals were not induced and served as the naïve control. Group 1 animals received three topical induction applications, one per week for three weeks. Skin reactions of the animals in Group 1 were recorded 24 and 48 hours following patch removal. Based on the results of the induction applications, 100% was selected as the highest non-irritating concentration for the challenge and was administered to both groups two weeks after the third induction. The skin reactions of all animals were recorded at 24 and 48 hours following patch removal. Body weights were recorded pretest and at termination and all animals were observed once per day for mortality, toxicity, and systemic effects. 85% hexylcinnemaldehyde (HCA) was the historical positive control with its toxicity confirmed in the laboratory every six months.
No mortality was observed and all animals gained body weight through the study period. Soiling and yellowing of the anogenital area was observed in treated as well as control animals. Erythema, absent to very faint was observed subsequent to the 1st induction while it was absent to moderate subsequent to the 2nd and 3rd inductions. Post challenge, absent to faint erythema was observed in the induced animals while absent to very faint erythema was observed in the naïve control group.
Based on the results observed, Santicizer P1700 was determined to be a dermal sensitizer in the under the conditions of this delayed contact dermal sensitization test in guinea pigs.
- Endpoint:
- skin sensitisation: in vitro
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available
- Justification for type of information:
- JUSTIFICATION FOR DATA WAIVING
Adequate in vivo skin sensitisation data is also available for assessment from studies conducted to fulfill requirements as mandated by the United States Environmental Protection Agency (US EPA).
Referenceopen allclose all
Table 1. Preliminary Test: Individual Dermal Irritation Scores |
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Animal Number |
Sex |
Day |
|||||||
1 |
2 |
3 |
6 |
||||||
Left |
Right |
Left |
Right |
Left1 |
Right1 |
Left |
Right |
||
Group 1P – 100%2 |
|||||||||
3680 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
1/0 |
1/0 |
0/0 |
0/0 |
3681 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
1 Oily fur around the dose site
2 25 µL of the test substance was applied as received to each ear (50 µL total)
Table 2. Preliminary Test: In-life Observations |
||||||||||
Animal Number |
Sex |
Group |
Dose Conc. (%) |
Observation |
Day of Observation (x = observation is present) |
|||||
1 |
2 |
3 |
4 |
5 |
6 |
|||||
3680 |
F |
1P |
100 |
Active and Healthy |
X |
X |
X |
X |
X |
X |
3681 |
F |
1P |
100 |
Active and Healthy |
X |
X |
X |
X |
X |
X |
Table 3. Main Test: Individual Body Weights |
||||
Animal Number |
Group |
Sex |
Day 1 (g) |
Day 6 (g) |
3601 |
1 Vehicle Control (AOO) |
Female |
21.9 |
22.0 |
3602 |
Female |
22.8 |
22.6 |
|
3603 |
Female |
20.5 |
21.0 |
|
3604 |
Female |
22.6 |
22.3 |
|
3605 |
Female |
21.6 |
21.8 |
|
|
||||
3606 |
2 Positive Control (25% HCA in AOO) |
Female |
22.0 |
22.3 |
3607 |
Female |
20.9 |
21.2 |
|
3608 |
Female |
20.9 |
21.0 |
|
3609 |
Female |
22.4 |
22.0 |
|
3610 |
Female |
21.8 |
21.9 |
|
|
||||
3611 |
3 10% Test Substance in AOO |
Female |
22.4 |
22.0 |
3612 |
Female |
24.6 |
23.0 |
|
3613 |
Female |
20.7 |
21.0 |
|
3614 |
Female |
20.2 |
20.8 |
|
3615 |
Female |
19.9 |
20.3 |
|
|
||||
3616 |
4 25% Test Substance in AOO |
Female |
25.3 |
23.9 |
3617 |
Female |
24.0 |
23.5 |
|
3618 |
Female |
23.0 |
23.2 |
|
3619 |
Female |
20.8 |
21.3 |
|
3620 |
Female |
20.2 |
20.8 |
|
|
||||
3621 |
5 50% Test Substance in AOO |
Female |
19.1 |
19.8 |
3622 |
Female |
23.5 |
22.7 |
|
3623 |
Female |
21.6 |
21.8 |
|
3624 |
Female |
20.4 |
20.9 |
|
3625 |
Female |
22.2 |
22.3 |
|
|
||||
3626 |
6 100% Test Substance in AOO |
Female |
22.6 |
23.1 |
3627 |
Female |
22.0 |
22.1 |
|
3628 |
Female |
21.0 |
21.4 |
|
3629 |
Female |
20.5 |
20.6 |
|
3630 |
Female |
20.9 |
21.4 |
|
Table 4. Main Test: Individual Dermal Irritation Scores (Erythema/Edema) |
|||||||||
Animal Number |
Sex |
Day |
|||||||
1 |
2 |
3 |
6 |
||||||
Left |
Right |
Left |
Right |
Left |
Right |
Left |
Right |
||
Group 1 – Vehicle Control1 |
|||||||||
3601 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3602 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3603 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3604 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3605 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
Group 2 - Positive Control2 |
|||||||||
3606 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
1/0 |
1/0 |
0/1 |
0/1 |
3607 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
1/0 |
1/0 |
1/0 |
1/0 |
3608 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
1/0 |
1/0 |
1/0 |
1/0 |
3609 |
F |
0/0 |
0/0 |
1/0 |
1/0 |
1/0 |
1/0 |
0/0 |
0/0 |
3610 |
F |
0/0 |
0/0 |
1/0 |
1/0 |
1/0 |
1/0 |
0/0 |
0/0 |
Group 3 – 10% Test Material3 |
|||||||||
3611 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3612 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3613 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3614 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3615 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
Group 4 – 25% Test Material3 |
|||||||||
3616 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3617 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3617 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3619 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3620 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
Group 5 – 50% Test Material3 |
|||||||||
3621 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3622 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3623 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3624 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3625 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
Group 6 – 100% Test Material4 |
|||||||||
3626 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3627 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3628 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3629 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
3630 |
F |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
0/0 |
1 25 µL of AOO was applied to each ear (50 µL total)
2 25 µL of a 25% w/w mixture of HCA in AOO was applied to each ear (50 µL total)
3 25 µL of the test material was applied as a w/w mixture in AOO to each ear (50 µL total)
4 25 µL of the test material was applied as received to each ear (50 µL total)
Table 5. Main Test: Individual and Mean DPM Values1 |
|||||||
Group |
Animal Number |
dpm |
dpm minus background2 |
Group Mean dpm minus background |
Std. Dev |
S.I3 |
SI ≥ 3 |
Background: 58.93 |
|||||||
1 Vehicle Control (AOO) |
3601 |
2280.09 |
2221.16 |
2711.08 |
666.63 |
- |
- |
3602 |
3829.58 |
3770.65 |
|||||
3603 |
2947.72 |
2888.79 |
|||||
3604 |
2624.33 |
2565.40 |
|||||
3605 |
2168.31 |
2109.38 |
|||||
|
|||||||
2 Positive Control (25% HCA in AOO) |
3606 |
12978.92 |
12919.99 |
15111.45 |
9874.68 |
5.57 |
Yes |
3607 |
14248.77 |
14189.84 |
|||||
3608 |
8715.97 |
8657.04 |
|||||
3609 |
32139.30 |
32080.37* |
|||||
3610 |
7768.92 |
7709.99 |
|||||
|
|||||||
3 10% Test Substance in AOO |
3611 |
2114.94 |
2056.01 |
1728.88 |
239.67 |
0.64 |
No |
3612 |
1610.14 |
1551.21 |
|||||
3613 |
1934.07 |
1875.14 |
|||||
3614 |
1754.14 |
1695.21 |
|||||
3615 |
1525.78 |
1466.85 |
|||||
|
|||||||
4 25% Test Substance in AOO |
3616 |
4821.19 |
4762.26 |
3081.15 |
1207.87 |
1.14 |
No |
3617 |
2031.04 |
1972.11 |
|||||
3617 |
2073.97 |
2015.04 |
|||||
3619 |
3897.53 |
3838.60 |
|||||
3620 |
2876.69 |
2817.76 |
|||||
|
|||||||
5 50% Test Substance in AOO |
3621 |
2872.26 |
2813.33 |
3275.35 |
742.81 |
1.21 |
No |
3622 |
2663.18 |
2604.25 |
|||||
3623 |
4410.35 |
4351.42 |
|||||
3624 |
3800.79 |
3741.86 |
|||||
3625 |
2924.80 |
2865.87 |
|||||
|
|||||||
6 100% Test Substance in AOO |
3626 |
6500.07 |
6441.14* |
2945.76 |
2027.63 |
1.09 |
No |
3627 |
1650.66 |
1591.73 |
|||||
3628 |
1535.79 |
1476.86 |
|||||
3629 |
2740.35 |
2681.42 |
|||||
3630 |
2596.57 |
2537.64 |
|||||
* The dpm values for Animal Nos.3609 and 3626 were determined to be outliers, but were included in the average and the standard deviation calculation for the test groups. There was no indication in the raw data that these animals were dosed incorrectly; therefore these animals were not excluded from the calculations.
1 Disintegrations per minute
2 Values analyzed for outliers, Grubbs1969.
3 Stimulation Index = Average dpm of Test Substance / Average dpm of Vehicle
Table 6. Main Test: Stimulation Index |
|||
Group |
Group Mean dpm |
SI |
Sensitization Response |
1 Vehicle Control (AOO) |
2711.08 |
- |
N/A |
2 Positive Control (25% HCA in AOO) |
15111.45 |
5.57 |
Positive – Valid Study |
3 10% Test Substance in AOO |
1728.88 |
0.64 |
Not a sensitizer |
4 25% Test Substance in AOO |
3081.15 |
1.14 |
Not a sensitizer |
5 50% Test Substance in AOO |
3275.35 |
1.21 |
Not a sensitizer |
6 100% Test Substance in AOO |
2945.76 |
1.09 |
Not a sensitizer |
N/A = Not applicable
Table 1. Results |
||||
Challenge |
Incidence Index |
Severity Index |
||
24 hours |
48 hours |
24 hours |
48 hours |
|
Santicizer P1700 |
0.15 |
0.20 |
0.20 |
0.25 |
Naïve Control |
0 |
0 |
0.05 |
0.05 |
Historical Positive Controla |
|
|||
HCA Positive Control |
0.40 |
0.40 |
0.48 |
0.48 |
Naïve Control |
0 |
0 |
0.20 |
0.15 |
a Historical Positive Control data (MB 15-23427.06) attached in Appendix B of the study report.
Table 2. Dermal Observations: Group 1 Post Patch Removal (Induction) |
||||||
Animal No. / Sex |
Induction 1, Erythema Site# 1 |
Induction 2, Erythema Site# 1 |
Induction 3, Erythema Site# 1 |
|||
24 hours |
48 hours |
24 hours |
48 hours |
24 hours |
48 hours |
|
D6838 / Male |
0 |
0 |
1 |
0 |
1 |
1 |
D6839 / Male |
0 |
0 |
0 |
0 |
0 |
0 |
D6840 / Male |
0 |
0 |
1 |
0.5 |
0 |
0 |
D6841 / Male |
0.5 |
0 |
0 |
0 |
1 |
0.5 |
D6842 / Male |
0.5 |
0.5 |
1 |
1 |
0 |
0 |
D6843 / Male |
0 |
0 |
1 |
1 |
1 |
1 |
D6844 / Male |
0 |
0 |
0 |
0 |
0 |
0 |
D6845 / Male |
0.5 |
0 |
0 |
0 |
1 |
1 |
D6846 / Male |
0 |
0 |
0 |
1 |
0 |
0 |
D6847 / Male |
0 |
0 |
1 |
1 |
1 |
1 |
|
||||||
D6848 / Female |
0 |
0 |
2 |
1 |
0 |
0 |
D6849 / Female |
0 |
0 |
1 |
1 |
0 |
1 |
D6850 / Female |
0 |
0 |
0 |
0 |
1 |
2 |
D6851 / Female |
0 |
0 |
1 |
1 |
0 |
1 |
D6852 / Female |
0 |
0 |
2 |
1 |
1 |
1 |
D6853 / Female |
0 |
0.5 |
1 |
2 |
1 |
1 |
D6854 / Female |
0 |
0 |
2 |
2 |
2 |
2 |
D6855 / Female |
0 |
0 |
1 |
1 |
1 |
2 |
D6856 / Female |
0 |
0 |
2 |
1 |
1 |
1 |
D6857 / Female |
0.5 |
0 |
2 |
1 |
1 |
2 |
Table 3. Dermal Observations: Groups 1 and 2 Post Patch Removal (Challenge) |
|||||
Animal No. / sex |
Induced Animals Site# 3 |
Animal No. / sex |
Naïve Control Site# 3 |
||
24 hours |
48 hours |
24 hours |
48 hours |
||
D6838 / Male |
0.5 |
0.5* |
D6858 / Male |
0 |
0 |
D6839 / Male |
0 |
0* |
D6859 / Male |
0 |
0.5* |
D6840 / Male |
0 |
0* |
D6860 / Male |
0 |
0 |
D6841 / Male |
0 |
0 |
D6861 / Male |
0 |
0 |
D6842 / Male |
0 |
0.5* |
D6862 / Male |
0 |
0 |
D6843 / Male |
0 |
0 |
|
|
|
D6844 / Male |
0 |
0* |
|
|
|
D6845 / Male |
1 |
1* |
|
|
|
D6846 / Male |
0 |
0* |
|
|
|
D6847 / Male |
0 |
0* |
|
|
|
|
|||||
D6848 / Female |
0 |
0 |
D6863 / Female |
0 |
0 |
D6849 / Female |
0 |
0 |
D6864 / Female |
0 |
0 |
D6850 / Female |
0 |
0* |
D6865 / Female |
0.5 |
0* |
D6851 / Female |
0 |
0 |
D6866 / Female |
0 |
0 |
D6852 / Female |
1 |
1* |
D6867 / Female |
0 |
0 |
D6853 / Female |
1 |
1* |
|
|
|
D6854 / Female |
0.5 |
1* |
|
|
|
D6855 / Female |
0 |
0* |
|
|
|
D6856 / Female |
0 |
0 |
|
|
|
D6857 / Female |
0 |
0* |
|
|
|
* Reclipped
Table 4. Body Weights (g): Groups 1 and 2 |
|||||
Animal No. / sex |
Pretest (g) |
Term (g) |
Animal No. / sex |
Pretest (g) |
Term (g) |
D6838 / Male |
362 |
650 |
D6858 / Male |
348 |
556 |
D6839 / Male |
356 |
640 |
D6859 / Male |
343 |
521 |
D6840 / Male |
363 |
612 |
D6860 / Male |
374 |
696 |
D6841 / Male |
318 |
596 |
D6861 / Male |
335 |
577 |
D6842 / Male |
353 |
594 |
D6862 / Male |
352 |
602 |
D6843 / Male |
322 |
529 |
Mean |
350 |
590 |
D6844 / Male |
339 |
591 |
S.D. |
14.64 |
66.08 |
D6845 / Male |
322 |
560 |
# |
5 |
5 |
D6846 / Male |
315 |
598 |
|
|
|
D6847 / Male |
318 |
522 |
|
|
|
Mean |
337 |
589 |
|
|
|
S.D. |
19.94 |
42.08 |
|
|
|
# |
10 |
10 |
|
|
|
|
|||||
D6848 / Female |
312 |
498 |
D6863 / Female |
332 |
536 |
D6849 / Female |
312 |
481 |
D6864 / Female |
312 |
468 |
D6850 / Female |
333 |
536 |
D6865 / Female |
332 |
560 |
D6851 / Female |
292 |
492 |
D6866 / Female |
330 |
434 |
D6852 / Female |
310 |
523 |
D6867 / Female |
317 |
556 |
D6853 / Female |
297 |
472 |
Mean |
325 |
511 |
D6854 / Female |
340 |
569 |
S.D. |
9.42 |
56.63 |
D6855 / Female |
288 |
487 |
# |
5 |
5 |
D6856 / Female |
315 |
489 |
|
|
|
D6857 / Female |
303 |
532 |
|
|
|
Mean |
310 |
508 |
|
|
|
S.D. |
16.63 |
30.71 |
|
|
|
# |
10 |
10 |
|
|
|
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
Two key studies were conducted on Santicizer Platinum P-1700 to determine the skin sensitisation potential, firstly a Buehler (3-induction) method in guinea pigs (MB Research Laboratories, 2015e) and then subsequently a mouse Local Lymph Node Assay (Product Safety Laboratories, 2020a). Whilst the guinea pig Buehler study (MB Research Laboratories, 2015e) resulted in a sensitising outcome, it is considered that the concentration used to challenge the guinea pigs (100%) was not the most appropriate. The test guidelines (OPPTS 870.2600 and OECD TG 406) state that the highest non-irritating concentration should be used for the challenge exposure. However, this study identified skin irritation effects based on dermal irritation scores in the induction phase (100%) of the study (skin irritation scores 0.5-2). No other pilot studies were conducted during the study to determine an appropriate non-irritating concentration for the challenge exposure therefore only a 100% concentration was used in the challenge with no consideration of a second challenge at a lower concentration. The observed skin irritation scores in the control and test animals challenged with 100% Santicizer Platinum P-1700 cannot be attributed to sensitisation alone as the concentration was considered to cause mild to moderate skin irritation. Therefore, this study is considered invalid and a second, more appropriate Local Lymph Node Assay study was conducted (Product Safety Laboratories, 2020).
This LLNA was conducted according to OECD TG 429 (2010) and OPPTS 870.2600 (2003) at appropriate testing concentrations (100, 50, 25 and 10%) using an appropriate vehicle acetone/olive oil; 4:1 v/v and positive (25% alpha hexylcinnamaldehyde) and negative (acetone/olive oil; 4:1 v/v) control groups. The LLNA performance was confirmed to be valid based on the sensitising outcome of the positive control (Stimulation Index (SI) 5.57). Therefore, concluding overall that under the conditions of the study Santicizer Platinum P-1700 is not considered to be a sensitizer (SI 1.09, 1.21, 1.14, 0.64).
Therefore, based on the available and reliable data, Santicizer Platinum P-1700 will not be classified for skin sensitisation.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on a weight of evidence from available substance specific data, P1700 does not meet the criteria for classification as a dermal sensitizer under EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
