Bis-O-(benzylidene)-D-glucitol

Endpoint summary

Description of key information

Acute Oral Toxicity:

The study was performed to assess the acute oral toxicity of the test item (EC 251-136-4) in the Wistar strain rat.

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

Results

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Body Weight. All animals showed expected gains in body weight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The acute oral median lethal dose (LD₅₀) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System-Unclassified).

Acute Dermal Toxicity:

The acute dermal toxicity of the substance (EC 251-136-4) has been assessed by reading across the results of studies conducted on two structurally similar analogue substances, which are detailed below.

Source Substance; Gel All DX (EC-413 -110 -2):

A study was performed to assess the acute dermal toxicity of the test material (EC 413-110-2) in the Sprague-Dawley CD strain rat.

A group of 10 animals (5 males and 5 females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for 14 days after the day of treatment and were then killed for gross pathological examination.

There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study.

All animals showed an expected gain in bodyweight during the study.

No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD₅₀) of the test material in the Sprague-Dawley CD strain rat was found to be > 2000 mg/kg bodyweight.

Source Substance; Gel All MD (EC 402-950-5):

The purpose of the study is to determine the acute dermal toxicity of technical GEL-ALL-MD (EC 402-950-5) to the rat.

Groups of 5 male and 5 female Sprague-Dawley rats received a single, 24 hour occluded, topical application of 2100 mg Technical Gel-ALL-MD/kg bodyweight, moistened with 0.5% w/v sodium carboxymethylcellulose in distilled water. A further 5 male and 5 female control animals were treated similarly except that no test material was applied to the skin. Animals were observed for 14 days after treatment and then examined post mortem.

There were no mortalities, no treatment-related clinical signs and no evidence of skin irritation. No treatment-related effects on bodyweight were recorded and no abnormalities were seen post-mortem.

In both sexes the acute dermal LD50 was greater than 2100 mg/kg bw.

Acute inhalation toxicity:

Study waived for target substance (EC 251-136-4).

An acute inhalation study conducted on source substance EC 402-950-5 gave an LC50 (4 hr) >0.67 mg/L air, the maximum attainable concentration.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental start date: 13 March 2017. Experimental completion date: 29 March 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

Identification: Geniset D
Batch: 5434
Purity: 99.4%
Physical state/Appearance: white powder
Expiry Date: 31 October 2018
Storage Conditions: room temperature in the dark

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Animal Information
Female Wistar (RccHan™:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non pregnant. After an acclimatization period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were 8 to 12 weeks of age. The body weight variation did not exceed ±20% of the mean body weight at the start of treatment.

Animal Care and Husbandry
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon,
UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

Test Item Preparation and Analysis:
For the purpose of the study the test item was freshly prepared, as required, as a suspension in arachis oil BP. Arachis oil BP was used because the test item did not dissolve/suspend in distilled water.
The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration.

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 females at 2000 mg/kg

Control animals:

no

Details on study design:

Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.
In the absence of toxicity at a dose level of 2000 mg/kg (single female), an additional group of animals was treated at 2000 mg/kg.

All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals.

Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.

Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.

At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and
opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted during the observation period.

Gross pathology:

No abnormalities were noted at necropsy

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System - Unclassified).

Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

Methods

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test item, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

Results

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Body Weight. All animals showed expected gains in body weight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion

The acute oral median lethal dose (LD₅₀) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg body weight (Globally Harmonized Classification System-Unclassified).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 October 1997 to 16 October 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: males: 215 to 243 g; females: 208 to 230 g
- Housing: animals were housed in suspended polypropylene cages furnished with wood flakes. The animals were housed individually during the 24-h exposure period and in groups of 5, by sex, for the remainder of the study.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 22 °C
- Humidity (%): 50 to 74 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/ 12
- On one occasion the relative humidity was above the limit specified in the protocol (70 %). This deviation was not considered to affect the purpose or integrity of the study.

Type of coverage:

semiocclusive

Vehicle:

other: moistened with arachis oil BP.

Details on dermal exposure:

TEST SITE
- % coverage: 10 % of the total body surface
- Type of wrap if used: surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage. The bandage was further secured with a piece of Blenderm wrapped around each end.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the treated skin and surrounding hair was wiped with cotton wool moistened with arachis oil BP to remoe any residual test material.
- Time after start of exposure: 24 h

TEST MATERIAL
- The test material, as received, was applied uniformly to an area of shorn skin which had been previously moistened with arachis oil BP.

Duration of exposure:

24 h

Doses:

2000 mg/kg

No. of animals per sex per dose:

5/ sex/ dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 h after dosing and subsequently once daily for 14 aysd. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: dermal reactions (scored according to the Draize scale).

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted during the study.

Gross pathology:

No abnormalities were noted at necropsy.

Other findings:

Dermal reactions: No signs of skin irritation were noted.

Interpretation of results:

not classified

Conclusions:

The dermal toxicity of the test material was assessed according to OECD guideline 402. The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley CD strain rat was found to be > 2000 mg/kg bodyweight.

Executive summary:

A study was performed to assess the actue dermal toxicity of the test material in the Sprague-Dawley CD strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 402 "Acute Dermal Toxicity" and Method B3 of Commission Directive 92/69/EEC.

A group of 10 animals (5 males and 5 females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for 14 days after the day of treatment and were then killed for gross pathological examination.

There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study.

All animals showed an expected gain in bodyweight during the study.

No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD₅₀) of the test material in the Sprague-Dawley CD strain rat was found to be > 2000 mg/kg bodyweight.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
It is proposed that the structural similarity and properties of the target substance and the structural analogue (sources substance) are sufficiently close for there to be a reasonable expectation of similar effects.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Soure substance:
IUPAC name: 1,3:2,4-bis-O-((3,4-dimethylphenyl)methylene) D-Glucitol
EC number: 413-110-2
CAS number: 135861-56-2


Target Substance: Bis-O-(benzylidene)-D-glucitol
IUPAC name: 1,3:2,4-bis-O-dibenzylidene-D-glucitol
EC number: 251-136-4
CAS number: 32647-67-9


3. ANALOGUE APPROACH JUSTIFICATION
Based on the structural similarity of the source substances and target substance, similarity of physic-chemical properties and similarity in experimental (eco)toxicological test data (and toxicokinetic behaviour assessment) it is concluded that target substance and the structural analogue (source substance) are sufficiently close for there to be a reasonable expectation of similar effects, for the endpoints where results have been read-across.

4. DATA MATRIX
Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

Reason / purpose for cross-reference:

read-across source

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted during the study.

Gross pathology:

No abnormalities were noted at necropsy.

Other findings:

Dermal reactions: No signs of skin irritation were noted.

Interpretation of results:

other: not classified under EU CLP

Conclusions:

The dermal toxicity of the test material was assessed according to OECD guideline 402. The acute dermal median lethal dose (LD50) of the test material in the Sprague-Dawley CD strain rat was found to be > 2000 mg/kg bodyweight.

Executive summary:

A study was performed to assess the actue dermal toxicity of the test material (structural analogue source substance EC 413-110-2) in the Sprague-Dawley CD strain rat. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 402 "Acute Dermal Toxicity" and Method B3 of Commission Directive 92/69/EEC.

A group of 10 animals (5 males and 5 females) was given a single 24-hour, semi-occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for 14 days after the day of treatment and were then killed for gross pathological examination.

There were no deaths. No signs of systemic toxicity or skin irritation were noted during the study.

All animals showed an expected gain in bodyweight during the study.

No abnormalities were noted at necropsy.

The acute dermal median lethal dose (LD₅₀) of the test material in the Sprague-Dawley CD strain rat was found to be > 2000 mg/kg bodyweight.

It is proposed that this result can be used in the assessment of the target substance.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study conducted between April 1988 and June 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes

Test type:

other: modified procedure described by Noakes and Sanderson (similar to standard acute method)

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK, Ltd, Manston Road, Margate, Kent
- Age at study initiation: Males were between 48 to 50 days old and females were between 56 to 65 days old
- Weight at study initiation: 278 g to 342 g for Males and 211 g to 267 g for Females
- Fasting period before study: not stated in report
- Housing: Rats were housed in pairs by sex and in steel mesh cages
- Diet (e.g. ad libitum): Free acess to Modified Expanded S.Q.C Rat and Mouse Diet No. 1
- Water (e.g. ad libitum): tap water
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):18 to 22 deg C
- Humidity (%): 44 to 54%
- Air changes (per hr): not stated in report
- Photoperiod (hrs dark / hrs light): 12 hours light followed by 12 hours of darkness

IN-LIFE DATES: From: Day of dosing (day 1) To: Day of sacrifice (day 14)

Type of coverage:

occlusive

Vehicle:

other: moistened with 0.5% sodium carboxymethyl in distilled water.

Details on dermal exposure:

TEST SITE
- Area of exposure: 6 cm by 10 cm
- Type of wrap if used: aluminium foil (held in place by an encircling band of waterproof plaster).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): treated skin was washed with soap and water to remove residual test material and then rinsed with water and dried.
- Time after start of exposure: 24 hours

TEST MATERIAL
- For solids, paste formed: yes, test material was moistened with 0.5% w/v solution sodium carboxymethylcellulose in distilled water and applied to the shaved area at a dose level of 2100 mg/kg.

Duration of exposure:

24 hours

Doses:

0 (control) and 2100 mg/kg

No. of animals per sex per dose:

5 males and 5 females were tested at both the 0 and 2100 mg/kg dose range (e.g. 20 animals in total)

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: Animals were observed for clinical signs and behaviour frequently on the day of treatment, at least once each morning and afternoon on working days thereafter and at leaast once each other day. Bodyweights were recorded immediately prior to treatment on Day 1, on Day 8 and immediately prior to termination on Day 15.

- Necropsy of survivors performed: yes - all animals were killed by CO2 asphyxiation on Day 15 and subjected to gross post-mortem examination for external abnormalities and for abnormalities of the thoracic and abdominal viscera.
- Other examinations performed: skin irritation - after removal of the plaster, skin treatment sites were examined daily for evidence of skin irritation.

Statistics:

The significance of difference between bodyweight change of control and test groups was estimated by the two sample test of Dunnett.

Preliminary study:

Range finding with 1 male and 1 female: no evidence of toxicity or skin irritation

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 100 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths during the study.

Clinical signs:

other: There were no treatment related clinical signs during the study.

Gross pathology:

There were no abnormal findings

Other findings:

Skin irritation: There was no evidence of skin irritation.

Interpretation of results:

not classified

Conclusions:

In both sexes, the acute dermal LD50 was greater than 2100 mg/kg bw.

Executive summary:

The purpose of the study is to determine the acute dermal toxicity of techncial GEL-ALL-MD to the rat.

Groups of 5 male and 5 female Sprague-Dawley rats received a single, 24 hour occluded, topical application of 2100 mg Technical Gel-ALL-MD/kg bodyweight, moistened with 0.5% w/v sodium carboxymethylcellulose in distilled water. A further 5 male and 5 female control animals were treated similarly except that no test material was applied to the skin. Animals were observed for 14 days after treatment and then examined post mortem.

There were no mortalities, no treatment-related clinical signs and no evidence of skin irritation. No treatment-related effects on bodyweight were recorded and no abnormalities were seen post-mortem.

In both sexes the acute dermal LD50 was greater than 2100 mg/kg bw.

Reference 4

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
It is proposed that the structural similarity and properties of the target substance and the structural analogue (sources substance) are sufficiently close for there to be a reasonable expectation of similar effects.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Soure substance:
IUPAC name: 1-(2,6-bis(4-tolyl)-1,3-dioxano(5,4-d)-1,3-dioxan-4-yl)ethane-1,2-diol
EC number: 402-950-5
CAS number: 87826-41-3

Target Substance: Bis-O-(benzylidene)-D-glucitol
IUPAC name: 1,3:2,4-bis-O-dibenzylidene-D-glucitol
EC number: 251-136-4
CAS number: 32647-67-9


3. ANALOGUE APPROACH JUSTIFICATION
Based on the structural similarity of the source substances and target substance, similarity of physic-chemical properties and similarity in experimental (eco)toxicological test data (and toxicokinetic behaviour assessment) it is concluded that target substance and the structural analogue (source substance) are sufficiently close for there to be a reasonable expectation of similar effects, for the endpoints where results have been read-across.

4. DATA MATRIX
Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

Reason / purpose for cross-reference:

read-across source

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 100 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths during the study.

Clinical signs:

other: There were no treatment related clinical signs during the study.

Gross pathology:

There were no abnormal findings

Other findings:

Skin irritation: There was no evidence of skin irritation.

Interpretation of results:

other: not classified under EU CLP

Conclusions:

In both sexes, the acute dermal LD50 was greater than 2100 mg/kg bw.

Executive summary:

The purpose of the study is to determine the acute dermal toxicity of techncial GEL-ALL-MD (structural analogue source substance EC 402-950-5) to the rat.

Groups of 5 male and 5 female Sprague-Dawley rats received a single, 24 hour occluded, topical application of 2100 mg Technical Gel-ALL-MD/kg bodyweight, moistened with 0.5% w/v sodium carboxymethylcellulose in distilled water. A further 5 male and 5 female control animals were treated similarly except that no test material was applied to the skin. Animals were observed for 14 days after treatment and then examined post mortem.

There were no mortalities, no treatment-related clinical signs and no evidence of skin irritation. No treatment-related effects on bodyweight were recorded and no abnormalities were seen post-mortem.

In both sexes the acute dermal LD50 was greater than 2100 mg/kg bw.

It is proposed that this result can be used in the assessment of the target substance.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Acute dermal toxicity study results are read-across from structurally similar analogue substances.

Justification for classification or non-classification

Acute toxicity: via oral route: The test material did not meet the criteria for classification as acutely toxic via the oral route according to Regulation 1272/2008 (CLP) as the LD₅₀ of >2000 mg/kg is above the threshold for classification.

Acute toxicity: via dermal route: The test material did not meet the criteria for classification as acutely toxic via the dermal route according to Regulation 1272/2008 (CLP) as the LD₅₀ of >2000 mg/kg is above the threshold for classification.