Reaction product of D-Glucopyranoside, methyl; esterified with oleic acid, methyl ester

Endpoint summary

Description of key information

Based on the results of the acute oral toxicity study (Weinberg, 2015(A)), the estimated LD50 of the test item was greater than 2000 mg/kg (CLP Category Unclassified) when administered once orally via gavage to fasted female albino rats.

Based on the results of the dermal acute toxicity study (Weinberg, 2015 (B)), the LD50 of the test item was greater than 2000 mg/kg (CLP category Unclassified) when administered once for 24 hours to the clipped, unabraded skin of male and female albino rats.

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:

2015-01-05 - 2015-01-27

Reliability:

1 (reliable without restriction)

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

The test substance was administered once orally via gavage to 1 fasted female albino rat at a dose level of 300 mg/kg. No mortality was observed, and another female rat was dosed at 2000 mg/kg (limit dose). No mortality was observed, and 4 additional female animals were dosed at 2000 mg/kg.
Mortality, clinical observations, and body weight changes were evaluated over a 14-day observation period. All animals were subjected to a gross necropsy.

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

The purity of the test substance was 100% (tested as formulated, 100% with no purity correction).
The test substance was stored at room temperature, protected from light, and was considered stable under these conditions. A reserve sample of the test substance was collected and stored in the WIL Research Archives.
Prior to use, the bulk test substance container was inverted and/or swirled. A sufficient amount of test substance was transferred into a storage container for dispensation.

Species:

rat

Strain:

other: Crl: CD(SD) albino rat

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Crl:CD(SD) rats from Charles River Laboratories, Inc., Raleigh, NC
- The number of animals selected was the minimum required to satisfy regulatory guidelines (5).
- No information exists to indicate a difference in sensitivity between the sexes. Therefore, only females were used to reduce variability and as means of minimizing the number of animals used.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks old
- Weight at study initiation: Approximately 170 to 300 grams at initiation of dosing, ± 20% of the mean of previously tested animals.
- Fasting period before study: fasted overnight before dosing
- Housing: individually in clean, stainless steel, wire-mesh cages suspended above cage-board
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 71 ± 5°F (22 ± 3°C)
- Humidity (%): 50 ± 20%
- Air changes (per hr): minimum of 10
- Photoperiod (hrs dark / hrs light): 12 hrs light and 12 hrs dark photoperiod

IN-LIFE DATES: From: 2014-12-30 To: 2015-01-27
All animals were euthanized at day 14 (end of the experimental phase).

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Remarks:

due to density of test item

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED:
Individual doses were calculated based on fasted body weights taken just prior to dosing and dose volumes of 0.31 and 2.04 mL/kg for the 300 and 2000 mg/kg groups, respectively.

The rats were fasted overnight prior to dosing. The test substance was administered orally via gavage. Food was returned approximately 4 hours after dosing.

Doses:

300 mg/kg bw
2000 mg/kg bw

No. of animals per sex per dose:

1 female at 300 mg/kg
4 females at 2000 mg/kg

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of clinical observations of mortality: ~15 minutes (± 5 minutes) and 1, 2 and 4 hours post-dosing on study day 0 and twice daily, once in the morning and once in the afternoon, thereafter for 14 days.
- Frequency of weighing: study days 0 (initiation), 7, and 14 (termination).
- Necropsy of survivors performed: yes (including major organ system of cranial, thoratic and abdominal cavities and the skin)
- Other examinations performed: clinical signs, body weight, mortality
Tissues were not collected

The test substance was administered once orally via gavage to 1 fasted female albino rat at a dose level of 300 mg/kg. No mortality was observed, and another female rat was dosed at 2000 mg/kg (limit dose). No mortality was observed, and 4 additional female animals were dosed at 2000 mg/kg.

Data Aquisition and Reporting
- Archive Management System (AMS): In-house developed application for storage, maintenance, and retrieval of information for archived materials (e.g., lab books, study data, wet tissues, slides, etc.).
- Formulations Dose Dispensing Management System (FDDMS): In-house developed system used to assign unique barcodes to formulation containers and individual containers used for dispensing dosing formulations.
- InSight® Publisher: Electronic publishing system (output is Adobe Acrobat, PDF).
- Master Schedule: Maintains the master schedule for the company.
- Metasys DDC Electronic Environmental Control System: Controls and monitors animal room environmental conditions.
- Microsoft® Office 2007 and 2010: Used in conjunction with the publishing software to generate study reports.
- Provantis Dispense™: Comprehensive system (Instem LSS Limited) to manage test materials, including receipt, formulation instructions, and accountability.
- WIL Metasys: In-house developed system used to record and report animal room environmental conditions.
- WIL Toxicology Data Management System™ (WTDMS™): In-house developed system used for collection and reporting of in-life and postmortem data.

Preliminary study:

No effects at dose 300 mg/kg when administered to one female albino rat in a single dose.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived to the scheduled necropsy.

Clinical signs:

There were no clinical findings observed during the study.

Body weight:

There were no remarkable body weight changes noted during the study (Table 1 - Table 4).

Gross pathology:

There were no macroscopic findings at the scheduled necropsy.

Other findings:

none

Table 1: Acute oral toxicity study in Albino rats, Summary of body weights [g]

Day	Group	300 mg/kg Female	2000 mg/kg Females
0	Mean S.D. N	192 0.0 1	199 9.2 5
7	Mean S.D. N	239 0.0 1	236 15.0 5
14	Mean S.D. N	259 0.0 1	251 15.1 5

Table 2: Acute oral toxicity study in Albino rats, Summary of bofy weight changes [g]

Day	Group	300 mg/kg Female	2000 mg/kg Females
0 to 7	Mean S.D. N	47 0.0 1	37 7.1 5
7 to 14	Mean S.D. N	20 0.0 1	15 6.7 5

Interpretation of results:

GHS criteria not met

Remarks:

limit dose at 2000 mg/kg

Conclusions:

Based on the results of this study, the estimated LD50 of the test item was greater than 2000 mg/kg (Unclassified for CLP) when administered once orally via gavage to fasted female albino rats.

Executive summary:

The acute oral toxicity of the test item was evaluated in this single-dose study in rats according to OECD Guideline 420 and in compliance with GLP.

The test substance was administered once orally via gavage to one fasted female albino rat at a dose level of 300 mg/kg. No mortality was observed, and another female rat was dosed at 2000 mg/kg (limit dose). No mortality was observed, and 4 additional female

animals were dosed at 2000 mg/kg.

Mortality, clinical observations, and body weight changes were evaluated over a 14-day observation period. All animals were subjected to a gross necropsy.

There were no deaths, remarkable body weight changes, or gross necropsy findings. No test substance-related clinical observations were noted.

Based on the results of this study, the estimated LD₅₀ of the test item was greater than 2000 mg/kg (CLP Category Unclassified) when administered once orally via gavage to fasted female albino rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

High qualitiy due to well performed test in accordance with GLP and official OECD Guideline. One supporting study was conducted also in compliance with GLP and principal international accepted methods.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-12-30 - 2015-01-20

Reliability:

1 (reliable without restriction)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

Prior to use, the bulk test substance container was inverted and/or swirled. A sufficient amount of test substance was transferred into a storage container for dispensation.

Species:

rat

Strain:

other: Crl:CD(SD)

Remarks:

albino rat

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approximately 9 weeks old
- The number of animals selected was the minimum required to satisfy regulatory guidelines.
- Weight at study initiation: ranges from 295 g to 302 g for males and 200 g to 215 g for fmales (± 20% of the mean for each sex).
- Housing: individually in clean, stainless steel, wire-mesh cages suspended above cage-board.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: minimum 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 70.4°F to 70.5°F (21.3°C to 21.4°C)
- Humidity (%): 32.0% to 43.0%
- Air changes (per hr): minimum 10
- Photoperiod (hrs dark / hrs light): 12 hrs light / 12 hrs dark

IN-LIFE DATES: From: 2014-12-30 To: 2015-01-20
All animals were euthanized at the end of the experimental period.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: maximum area possible on the dorsal skin.
- % coverage: at least 10 % of the total surface
- Type of wrap if used: gauze bandages (<8 ply) secured with nonirritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): wiped with disposable paper towels moistened with tepid tap water
- Time after start of exposure: removal 24 hrs following test substance application

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):Individual doses were calculated based on body weights taken just prior to dosing and a dose volume of 2.04 mL/kg.
- Concentration (if solution): 100 %
- Constant volume or concentration used: yes

Duration of exposure:

24 hrs

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 females at 2000 mg/kg
5 males at 2000 mg/kg

Control animals:

no

Details on study design:

One group of 5 male and 5 female rats were dermally administered a single dose (24-hour, semi-occluded exposure) of the test item at a dose level of 2000 mg/kg (limit dose). On the day prior to dosing, the hair was removed from the backs and flanks of the rats using a small animal clipper.
- Duration of observation period following administration: 14 days
- Frequency of clinical observations and mortality:approximately 1, 2, and 4 hours post-application on study day 0 and once daily thereafter for 14 days
- Frequency of weighing: on study days 0 (initiation), 7, and 14 (termination).
- Frequency of dermal observations: The application sites were examined for erythema, edema (Draize, 1965), and other dermal findings beginning 30-60 minutes after bandage removal and daily thereafter through study day 14. The areas of application were clipped free of hair on the day prior to dosing and as needed to facilitate accurate dermal observations.
- Necropsy (of the major organ systems of the cranial, thoracic, and abdominal cavities and the skin) of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights. Tissues were not collected. Observations included, but were not limited to, evaluation for changes in appearance of skin and fur, eyes, mucous membranes, respiratory and circulatory systems, autonomic effects, and central nervous system effects

DATA ACQUISITION AND REPORTING
Program/System Description / Description
Archive Management System (AMS): In-house developed application for storage, maintenance, and retrieval of information for archived materials (e.g., lab books, study data, wet tissues, slides, etc.).
Formulations Dose Dispensing Management System (FDDMS): In-house developed system used to assign unique barcodes to formulation containers and individual containers used for dispensing dosing formulations.
InSight® Publisher: Electronic publishing system (output is Adobe Acrobat, PDF).
Master Schedule: Maintains the master schedule for the company
Metasys DDC Electronic Environmental Control System: Controls and monitors animal room environmental conditions
Microsoft® Office 2007 or higher: Used in conjunction with the publishing software to generate study reports.
Provantis Dispense™: Comprehensive system (Instem LSS Limited) to manage test materials, including receipt, formulation instructions, and accountability.
WIL Metasys: In-house developed system used to record and report animal room environmental conditions
WIL Toxicology Data Management System™ (WTDMS™): In-house developed system used for collection and reporting of in-life and postmortem data.

Preliminary study:

not performed

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

There were no deaths during the study.

Clinical signs:

There were no clinical findings observed during the study.

Body weight:

There were no remarkable body weight changes noted during the study.

Gross pathology:

There were no macroscopic findings at the scheduled necropsy.

Other findings:

Dermal Observations:
Dermal findings noted during the study consisted of very slight (grade 1) to slight (grade 2) erythema and desquamation for all 5 males and 5 females. Erythema subsided by study day 6, and desquamation subsided by study day 11. In addition, female nos. 5126 and 5129 had scabbing within the dose site on study day 2 and 3, respectively. There was no edema observed at any dose site.

Table1: Acute dermal toxicity study in Albino rats, Summary of clinical findings: Total occurence/No. of animals

Table Range: Group:	Male Day 0 to Day 14 1 (2000 mg/kg)
Normal
No significant clinical observations	5/5
Disposition
Scheduled euthanasia; primary (Day 14)	5/5
Acutes
Time of Dosing	5/5
Appeared normal	85/5
Scheduled euthanasia; primary (Day 14)	5/5
Dermal observations
Scored, not remarkable	42/5
No erythema	17/5
Erhythema - very slight	11/5
No edema	28/5
Desquamation	24/5

Table 2: Acute dermal toxicity study in Albino rats, Summary of clinical findings: Total occurence/No. of animals

Table Range: Group:	Female Day 0 to Day 14 1 (2000 mg/kg)
Normal
No significant clinical observations	5/5
Disposition
Scheduled euthanasia; primary (Day 14)	5/5
Acutes
Time of Dosing	5/5
Appeared normal	85/5
Scheduled euthanasia; primary (Day 14)	5/5
Dermal observations
Scored, not remarkable	37/5
No erythema	21/5
Erhythema - very slight	9/5
Erythema - slight	3/2
No edema	33/5
Desquamation	32/5
Scabbing within dose site	2/2

Table 4: Acute dermal toxicity study in Albino rats, Summary of body weights

Group: 2000 mg/kg Males
Day
0	Mean S.D. N	298 2.9 5
7	Mean S.D. N	332 2.3 5
14	Mean S.D. N	379 7.2 5
Group: 2000 mg/kg Females
Day
0	Mean S.D. N	205 6.0 5
7	Mean S.D. N	218 6.5 5
14	Mean S.D. N	239 11.2 5

Table 5: Acute dermal toxicity study in Albino rats, Summary of body weight changes

Group: 2000 mg/kg Males
Day
0 to 7	Mean S.D. N	35 3.4 5
7 to 14	Mean S.D. N	47 4.9 5
Group: 2000 mg/kg Females
0 to 7	Mean S.D. N	13 2.4 5
7 to 14	Mean S.D. N	21 9.3 5

Interpretation of results:

GHS criteria not met

Remarks:

limit dose 2000 mg/kg

Conclusions:

Based on the results of this study, the LD50 of the test item was greater than 2000 mg/kg (CLP category Unclassified) when administered once for 24 hours to the clipped, unabraded skin of male and female albino rats.

Executive summary:

The acute dermal median lethal dose (LD₅₀) of the test item was conducted according the EPA OPPTS Guideline 870.1200 and the OECD Guideline Section 402, in compliance with GLP.

The test item was administered once dermally for a 24-hour period under gauze dressing semiocclusive to the skin of Crl:CD(SD) albino rats to perform the limit test. The test substance was administered to one group of five male and five female rats at a dose level of 2000 mg/kg. Mortality, clinical observations, dermal findings (Draize, 1965), and body weight changes were evaluated over a 14-day observation period. All animals were subjected to a gross necropsy.

There were no deaths, clinical observations, remarkable body weight changes, or test substance-related gross necropsy findings. Dermal findings noted during the study consisted of very slight (grade 1) to slight (grade 2) erythema and desquamation for all five males and five females and scabbing within the dose site for two females.

Based on the results of this study, the LD₅₀ of the test item was greater than 2000 mg/kg (CLP category Unclassified) when administered once for 24 hours to the clipped, unabraded skin of male and female albino rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

High qualitiy due to well performed test in accordance with GLP and official OECD Guideline.

Additional information

Acute oral toxicity data:

In the key study (Weinberg, 2015 (A)), the acute oral toxicity of the test item was evaluated in this single-dose study in rats according to OECD Guideline 420 and in compliance with GLP.

The test substance was administered once orally via gavage to one fasted female albino rat at a dose level of 300 mg/kg. No mortality was observed, and another female rat was dosed at 2000 mg/kg (limit dose). No mortality was observed, and four additional female animals were dosed at 2000 mg/kg. Mortality, clinical observations, and body weight changes were evaluated over a 14-day observation period. All animals were subjected to a gross necropsy.

There were no deaths, remarkable body weight changes, or gross necropsy findings. No test substance-related clinical observations were noted. Based on the results of this study, the estimated LD₅₀ of the test item was greater than 2000 mg/kg (CLP Category Unclassified) when administered once orally via gavage to fasted female albino rats.

In a supporting study (Smolin, 1983), a GLP compliant acute oral toxicity test in albino rats was performed. Ten (5M:5F) albino rats, 208 - 278 g, each received a single oral dose of the test item at a dose level of 5000 mg/kg body weight. Animals were observed for pharmacologic activity and drug toxicity 1, 3, 6, and 24 hours after treatment, and daily therafter for a total of 14 days. Non-survivors and animals surviving the 14 day observation period were subjected to gross necropsy, with all findings noted. The test article was used as received.

The test item is not orally toxic to rats under conditions of this test.

Acute dermal toxicity data:

In the GLP- key study (Weinberg, 2015 (B)) the acute dermal median lethal dose (LD₅₀) of the test item was conducted according the EPA OPPTS Guideline 870.1200 and the OECD Guideline Section 402. The test item was administered once dermally for a 24-hour period under gauze dressing semiocclusive to the skin of Crl:CD(SD) albino rats to perform the limit test. The test substance was administered to one group of five male and five female rats at a dose level of 2000 mg/kg. Mortality, clinical observations, dermal findings (Draize, 1965), and body weight changes were evaluated over a 14-day observation period. All animals were subjected to a gross necropsy. There were no deaths, clinical observations, remarkable body weight changes, or test substance-related gross necropsy findings. Dermal findings noted during the study consisted of very slight (grade 1) to slight (grade 2) erythema and desquamation for all five males and five females and scabbing within the dose site for two females.

Based on the results of this study, the LD₅₀ of the test item was greater than 2000 mg/kg (CLP category Unclassified) when administered once for 24 hours to the clipped, unabraded skin of male and female albino rats.

Acute inhalation toxicity:

The test item is a liquid with a vapour pressure of 0.00627 Pa at 25 °C (Tremain, 2015) and is used primarily as a component of lubricants and greases by workers, professionals and consumers. It is expected that inhalation exposure from these uses will be low and that the most likely route of exposure for workers and consumers is the dermal route.Therefore, testing for inhalation toxicity is not scientifically justified (see waiver). The potential for the generation of inhalable forms is low, also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route of exposure is not likely. In accordance with REACH Regulation, Annex VIII, section 8.5.2, column two, testing for acute inhalation toxicity can be waived.

Justification for classification or non-classification

The key studies for oral acute toxicity of the test item reveals an acute oral LD₅₀ of greater than 2000 mg/kg bw in rats. The key study for dermal acute toxicity of the test item shows an acute dermal LD₅₀ of greater than 2000 mg/kg bw in rats.

Testing for acute inhalation toxicity can be waived in accordance with REACH Regulation, Annex VIII, section 8.5.2, column two.

Therefore the test item does not meet the criteria for classification and labelling for oral, dermal or inhalatory toxicity in accordance with the Regulation (EC) No 1272/2008 (CLP Regulation).