Reaction mass of octan-2-ylbenzene, octan-3-ylbenzene and octan-4-ylbenzene

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
According to conditions described in Column 2 of Section 8.5.3 of Annex VIII testing by the dermal route is appropriate if: (1) inhalation of the substance is unlikely; and (2) skin contact in production and/or use is likely; and (3) the physicochemical and toxicological properties suggest potential for a significant rate of absorption through the skin. Since SHR 1396 is only used in hermetically sealed compressors skin contact in production and/or use is unlikely to result in exposure to SHR 1396. Since SHR 1396 is expected to have limited absorption through the skin based on its low water solubility (0.04 mg/L) and high Log Pow (7.0 – 7.2), showed no systemic effects in an in vivo study (LLNA) with dermal exposure and has already low acute oral toxicity it is unlikely that the acute dermal toxicity will exceed the oral toxicity. Since criteria (2) and (3) in Column 2 of section 8.5.3 of Annex VIII are not fulfilled, testing by the dermal route for SHR 1396 is considered not appropriate and thus testing is waived.

See Cross references for relevant supporting data on uses, toxicological data and physical and chemical properties.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 Jul - 06 Aug 2019

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:

2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Version / remarks:

2003

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

2012

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France.
- Females nulliparous and non-pregnant: yes.
- Microbiological status of animals: SPF-quality.
- Age at study initiation: 10 weeks old.
- Weight at study initiation: 19.6 to 23.0 grams.
- Housing: Animals were group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum.
- Water: Municipal tap-water was freely available to each animal via water bottles.
- Acclimation period: at least 5 days.
- Indication of any skin lesions: no.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): The daily mean temperature during the study period was 22 to 23°C.
- Humidity (%): The daily mean relative humidity of 53 to 79%.
- Air changes (per hr): at least 10.
- Photoperiod (hrs dark / hrs light): 12/12
- IN-LIFE DATES: From 17 Jul to 05 Aug.

Vehicle:

acetone/olive oil (4:1 v/v)

No. of animals per dose:

5 animals

Details on study design:

PRE-SCREEN TESTS:
Two test item concentrations were tested; a 50% (w/w) and 100% (w/w) concentration.
Two young adult females per concentration were selected. Each animal was treated with one concentration on three consecutive days. Ear thickness measurements were conducted using a digital thickness gauge (Kroeplin C110T-K) prior to dosing on Days 1 and 3, and on Day 6. Animals were sacrificed after the final observation.

MAIN STUDY

Three groups of five animals were treated with one test item concentration per group. The highest test item concentration was selected from the pre-screen test. One group of five animals was treated with the vehicle.
Group 1 was treated with the vehicle (vehicle control group). Group 2, 3 and 4 were treated with 25%, 50% and 100% concentration, respectively.

TREATMENT PREPARATION AND ADMINISTRATION:

Test item dosing formulations (w/w) were homogenized to visually acceptable levels at appropriate concentrations to meet dose level requirements.
The dosing formulations were prepared daily and dosed within 4 hours after adding the vehicle to the test item.
The dosing formulations were kept at room temperature until dosing.
The dosing formulations were stirred until and during dosing.
No adjustment was made for specific gravity of the vehicle and no correction was made for the purity/composition of the test item.
- Induction: Test item (25 μL) was applied in the dorsal surface of both ears of each animal, for 3 consecutive days.
- Excision of the Nodes. In day 6, each animal was injected via the tail vein with 0.25 mL of sterile phosphate buffered saline (PBS) containing 20 μCi of 3H-methyl thymidine. After five hours, all animals were euthanized by intraperitoneal injection (0.2 mL/animal) of Euthasol® 20%. The draining (auricular) lymph node of each ear was excised. The relative size of the nodes (as compared to normal) was estimated by visual examination and abnormalities of the nodes and surrounding area were recorded. The nodes were pooled for each animal in PBS.
- Tissue processing for radioactivity: On day 6 and following the excision of the nodes, a single cell suspension of limph node cells was prepared in PBS. LNC were washed twice and DNA was precipitated using 5% trichloroacetic acid (TCA).
- Radioactivity measurements: On day 7, precipitates were recovered by centrifugation, resuspended in 1 mL TCA and transferred to 10 mL of Ultima Gold cocktail as the scintillation fluid. Radioactivity measurements were performed using a Packard scintillation counter (2910TR). Counting time was to a statistical precision of ± 0.2% or a maximum of 5 minutes whichever came first. The scintillation counter was programmed to automatically subtract background and convert Counts Per Minute (CPM) to Disintegrations Per Minute (DPM).

ANALYSIS
DPM values are presented for each animal and for each dose group. A Stimulation Index (SI) is calculated for each group using the individual SI values. The individual SI is the ratio of the DPM/animal compared to the DPM/vehicle control group mean.

INTERPRETATION
If the results indicate a SI ≥ 3, the test item may be regarded as a skin sensitizer.
The results were evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2017) (including all amendments) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of items and mixtures, including all amendments.
Consideration was given to the EC3 value (the estimated test item concentration that will give a SI =3) (see table 1 in 'any other information on material and methods')

Key result

Parameter:

SI

Value:

1.9

Test group / Remarks:

25%

Key result

Parameter:

SI

Value:

2.4

Test group / Remarks:

50%

Key result

Parameter:

SI

Value:

2.5

Test group / Remarks:

100%

Cellular proliferation data / Observations:

PRE-SCREENING TEST:
- At a 50% and 100% test item concentration, no signs of systemic toxicity were noted and up to very slight erythema was observed and therefore the 100% concentration was selected as highest concentration for the main study.

MAIN STUDY

SKIN REACTIONS / IRRITATION:
The very slight erythema of the ears as shown by all test item treated animals on at 1 to 6 Days during the observation period was considered not to have a toxicologically significant effect on the activity of the nodes.

CLINICAL OBSERVATIONS: No mortality occurred and no clinical signs of systemic toxicity were observed in the animals.

BODY WEIGHTS: Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

MACROSCOPIC EXAMINATION OF LyMPH NODES:
The majority of auricular lymph nodes were considered normal in size, except for the nodes in two animals treated at 100%, which were considered to be slightly enlarged.
No macroscopic abnormalities of the surrounding area were noted for the majority of animals. Pale discoloration of the surrounding area of the lymph nodes of the animals treated at 100% was noted, which was considered not to have affected the DPM values of these animals.

DETAILS ON STIMULATION INDEX CALCULATION : DPM (Disintegrations Per Minute) values are presented for each animal and for each dose group. A Stimulation Index (SI) is calculated for each group using the individual SI values. The individual SI is the ratio of the DPM/animal compared to the DPM/vehicle control group mean.

Interpretation of results:

other: Not skin sensitising

Remarks:

According to Regulation (EC) No. 1272/2008 and its amendments.

Conclusions:

In conclusion, since there is no indication that the test item elicited a SI ≥ 3 when tested up to 100%, SHR 1396 was considered not to be a skin sensitizer.

Executive summary:

A LLNA study was performed following OECD guidelines and GLP principles. Three experimental groups of five females were treated with test item concentrations of 15%, 50% and 100%. One control group of five females was treated with the vehicle (Acetone/olive oil 1:4 v/v). All animals were treated for 3 consecutive days. Three days after the last exposure all animals were injected with 3H-methyl thymidine for measuring radioactivity. The activity was expressed as the number of disintegrations per minute (DPM) and a stimulation index (SI) was calculated for each group. The six-month reliability check with Alpha-hexylcinnamaldehyde indicates that the Local Lymph Node Assay as performed at Charles River Den Bosch is an appropriate model for testing for contact hypersensitivity. No macroscopic abnormalities of the surrounding area were noted for the majority of animals. Pale discoloration of the surrounding area of the lymph nodes of the animals treated at 100% was noted, which was considered not to have affected the DPM values of these animals. Mean DPM/animal values for the experimental groups treated with test item concentrations 25, 50 and 100% were 717, 938 and 972 DPM, respectively. The mean DPM/animal value for the vehicle control group was 384 DPM. The SI values calculated for the test item concentrations 25, 50 and 100% were 1.9, 2.4 and 2.5, respectively. Since there was no indication that the test item elicited a SI ≥ 3 when tested up to 100%, SHR 1396 was considered not to be a skin sensitizer. Based on these results, SHR 1396 would not be regarded as a skin sensitizer according to the recommendations made in the test guidelines. The test item does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Cross-reference 2

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

1.18 mg/m³

Most sensitive endpoint:

effect on fertility

Route of original study:

Oral

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

150

Dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Modified dose descriptor starting point:

NOAEC

Value:

176 mg/m³

Explanation for the modification of the dose descriptor starting point:

Kinetics (metabolism, distribution and excretion) are considered to be similar for oral and inhalation intake in the absence of evidence of the opposite.

Starting point: NOAEL of 100 mg/kg bw/day in a 28-day combined repeated dose repro screener toxicity study in rat.

Conversion of an oral NOAEL into a corrected NOAEC:

For workers (8h exposure/day), the corrected inhalatory NOAEC = oral NOAEL * 1/sRVrat * ABSoral-rat /ABSinhal-human * sRVhuman/wRV

          = 100 * 1/0.38 m³/kg/8h * ABSoral-rat /ABSinhal-human * 6.7 m³(8h)/10 m³(8h)

          = 100 * 1/0.38 m³/kg/8h * 1 * 6.7 m3 (8h)/10 m³(8h)

= 100 /0.38 * 1* (6.7/10) = 176.3 mg/m³.

 

With ABS: Absorption, sRV: Standard Respiratory Volume; wRV: Worker Respiratory Volume; ABSoral-rat /ABSinhal-human= 50/50= 1, assuming no differences in inhalation absorption between rats and humans.

AF for dose response relationship:

2

Justification:

Value is NOAEL. An additional factor 2 is included to take account of the lower sensitivity the OECD 422 study for fertility effects.

AF for differences in duration of exposure:

6

Justification:

Default (DNEL calculator)

AF for interspecies differences (allometric scaling):

1

Justification:

Default (DNEL calculator)

AF for other interspecies differences:

2.5

Justification:

Default (DNEL calculator)

AF for intraspecies differences:

5

Justification:

Default (DNEL calculator)

AF for the quality of the whole database:

1

Justification:

Default (DNEL calculator)

AF for remaining uncertainties:

1

Justification:

No remaining uncertainties

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

0.833 mg/kg bw/day

Most sensitive endpoint:

effect on fertility

Route of original study:

Oral

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

600

Dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Modified dose descriptor starting point:

NOAEL

Value:

500 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Kinetics (metabolism, distribution and excretion) are considered to be 50% for oral and 10% for dermal intake.Correction dermal NOAEL: 50/10^a= 500 mg/kg bw/day (^a% oral/dermal absorption)

AF for dose response relationship:

2

Justification:

Value is NOAEL. An additional factor 2 is included to take account of the lower sensitivity the OECD 422 study for fertility effects

AF for differences in duration of exposure:

6

Justification:

Default (DNEL calculator)

AF for interspecies differences (allometric scaling):

4

Justification:

Default (DNEL calculator)

AF for other interspecies differences:

2.5

Justification:

Default (DNEL calculator)

AF for intraspecies differences:

5

Justification:

Default (DNEL calculator)

AF for the quality of the whole database:

1

Justification:

Default (DNEL calculator)

AF for remaining uncertainties:

1

Justification:

No remaining uncertainties

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

0.29 mg/m³

Most sensitive endpoint:

effect on fertility

Route of original study:

Oral

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

300

Dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Modified dose descriptor starting point:

NOAEC

Value:

87 mg/m³

Explanation for the modification of the dose descriptor starting point:

Kinetics (metabolism, distribution and excretion) are considered to be similar for oral and inhalation intake in the absence of evidence of the opposite.

Starting point: NOAEL of 100 mg/kg bw/day in a 28-day combined repeated dose repro screener toxicity study in rat.

Conversion of an oral NOAEL into a corrected NOAEC:

For general population (24h exposure/day), the corrected inhalatory NOAEC = oral NOAEL * 1/sRVrat * ABSoral-rat /ABSinhal-human

= 100 * 1/1.15 m³/kg/d * ABSoral-rat /ABSinhal-human

= 100 * 1/1.15 m³/kg/d * 1

= 100 /1.15 * 1 = 87.0 mg/m³

 

With ABS: Absorption, sRV: Standard Respiratory Volume; ABSoral-rat /ABSinhal-human= 50/50= 1, assuming no differences in inhalation absorption between rats and humans.

AF for dose response relationship:

2

Justification:

Value is NOAEL. An additional factor 2 is included to take account of the lower sensitivity the OECD 422 study for fertility effects.

AF for differences in duration of exposure:

6

Justification:

Default (DNEL calculator)

AF for interspecies differences (allometric scaling):

1

Justification:

Default (DNEL calculator)

AF for other interspecies differences:

2.5

Justification:

Default (DNEL calculator)

AF for intraspecies differences:

10

Justification:

Default (DNEL calculator)

AF for the quality of the whole database:

1

Justification:

Default (DNEL calculator)

AF for remaining uncertainties:

1

Justification:

No remaining uncertainties

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

0.417 mg/kg bw/day

Most sensitive endpoint:

effect on fertility

Route of original study:

Oral

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

1 200

Dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Modified dose descriptor starting point:

NOAEL

Value:

500 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Kinetics (metabolism, distribution and excretion) are considered to be 50% for oral and 10% for dermal intake.Correction dermal NOAEL: 50/10^a= 500 mg/kg bw/day (^a% oral/dermal absorption)

AF for dose response relationship:

2

Justification:

Value is NOAEL. An additional factor 2 is included to take account of the lower sensitivity the OECD 422 study for fertility effects.

AF for differences in duration of exposure:

6

Justification:

Default (DNEL calculator)

AF for interspecies differences (allometric scaling):

4

Justification:

Default (DNEL calculator)

AF for other interspecies differences:

2.5

Justification:

Default (DNEL calculator)

AF for intraspecies differences:

10

Justification:

Default (DNEL calculator)

AF for the quality of the whole database:

1

Justification:

Default (DNEL calculator)

AF for remaining uncertainties:

1

Justification:

No remaining uncertainties

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

83.3 µg/kg bw/day

Most sensitive endpoint:

effect on fertility

Route of original study:

Oral

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

1 200

Dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Modified dose descriptor starting point:

NOAEL

Value:

100 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Default (DNEL calculator)

AF for dose response relationship:

2

Justification:

Value is NOAEL. An additional factor 2 is included to take account of the lower sensitivity the OECD 422 study for fertility effects.

AF for differences in duration of exposure:

6

Justification:

Default (DNEL calculator)

AF for interspecies differences (allometric scaling):

4

Justification:

Default (DNEL calculator)

AF for other interspecies differences:

2.5

Justification:

Default (DNEL calculator)

AF for intraspecies differences:

10

Justification:

Default (DNEL calculator)

AF for the quality of the whole database:

1

Justification:

Default (DNEL calculator)

AF for remaining uncertainties:

1

Justification:

No remaining uncertainties

Hazard assessment conclusion:

no hazard identified

Hazard assessment conclusion:

no hazard identified

Cross-reference 3

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Registration/ Notification status for the use:

use registered according to REACH Article 10; total tonnage manufactured/imported >=10tonnes/year per registrant

Use name:

Use at industrial site by the registrant

Name of activity / technique:

Use of the substance

Environmental release category (ERC):

ERC5: Use at industrial site leading to inclusion into/onto article

Name of activity / technique:

Use of the substance

Process category (PROC):

PROC 1: Chemical production or refinery in closed process without likelihood of exposure or processes with equivalent containment conditions

Substance supplied to that use in form of:

as such

Subsequent service life relevant for this use:

yes

Related assessment:

use assessed in an own CSR

Cross-reference 4

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

In an acute oral toxicity study with female rats, performed according to OECD 423 and GLP principles, a LD50 >2000 mg/kg bw was determined.

The determination of Acute inhalation toxicity and Acute Dermal toxicity is waived based on exposure considerations and the physicochemical and toxicological properties of SHR 1396.

Endpoint conclusion:

no adverse effect observed

Endpoint conclusion:

no study available

Endpoint conclusion:

no study available

An acute oral toxicity study was performed according to OECD 423 test guideline and GLP principles. SHR 1396 was administered by oral gavage to two consecutive groups of three female Wistar Han rats at 2000 mg/kg body weight. No mortality occurred. Hunched posture and/or piloerection was noted for all animals between Days 1 and 5. The mean body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals. Based on these results, a LD50 >2000 mg/kg body weight was determined.

Based on the current available information, the substance does not have to be classified for acute toxicity according to the CLP Regulation.