Tall oil, maleated

Endpoint summary

Description of key information

Based on the experimental study results, the acute oral LD50 in rats of the test item, Tall Oil, Maleated (EnvaMul™ 600) was found to be greater than 2000 mg/kg. Therefore, the test item is considered not to present a significant acute toxic risk if swallowed.

Acute inhalation toxicity study not conducted as exposure is not likely to occur.

The dermal LD50 value of EnvaMul 600 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, EnvaMul 600 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

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:

Study Initiation: October 8, 2013; Treatment Start: October 15, 2013; Experimental Completion: November 18, 2013; Study Completion: November 20, 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

up-and-down procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Test Animal Rattus norvegicus
Strain: CD® [Crl:CD®(SD)BR] Sprague-Dawley
Source: Charles River Canada, Montreal, Quebec
Number and Sex: 5 female rats were used.
Body Weight: Range 200.2 - 216.7 g after fasting. The weight variation in animals at the start of the study did not exceed ±20 percent of the mean weight.
Acclimatization Period: 7 days
Age at Study Start: Approximately 7 to 8 weeks
Animal Identification: Body colour coding, cage labels
Animal Housing and Maintenance: Female rats were individually housed in separate quarters in solid bottom cages. Individual animals were identified by colour coding; the animal number and group number also appeared on the outside of each cage to preclude mix-up. The animal room environment was controlled (targeted ranges: temperature 19 °C to 25 °C, relative humidity 30 - 70%, minimum 10 air changes per hour) and monitored. The photo-cycle was 12 hours light and 12 hours dark. Upon arrival all animals were submitted to a general physical examination and all were found healthy and were admitted. Teklad Certified Rodent Diet and water were offered ad libitum throughout the acclimatization and study periods, except as specified under “Preparation of Animals”.
The cage cleaning schedule, air filtration and recirculation, health checks and facility maintenance were carried out in accordance with the applicable Nucro-Technics’ Standard Operating Procedures, and such activities were recorded in the animal room records. Animals were housed and maintained according to the AAALAC International Guide for the Care and Use of Laboratory Animals, CCAC Guidelines for Care and Use of Experimental Animals and Nucro-Technics’ Standard Operating Procedures.
Animal Selection: The test population of animals was randomly selected from newly arrived, previously unused rats.
Preparation of Animals: All animals used for the Limit Test were fasted over-night. Food but not water was withheld beginning at 4:00 p.m. on the day preceding dosing, and was returned to the cages approximately 1 hour after dosing.

Route of administration:

oral: gavage

Vehicle:

not specified

Doses:

The first animal was dosed at 2000 mg/kg. Since this animal survived, four additional animals were sequentially dosed at 2-day intervals. A total of five animals were dosed.
The test animals were dosed at 2000 mg/kg (2.0 mL/kg by volume of test item).

No. of animals per sex per dose:

The individual doses of the test item were individually calculated for each animal based on the body weight of the animal. All doses were administered orally using a feeding cannula inserted into the stomach of the animals.

Control animals:

no

Details on study design:

Method
The method used for conducting this study is the accepted standard described in the Organisation for Economic Co-operation and Development (OECD) Guideline for the Testing of Chemicals, Acute Oral Toxicity (Up-and-Down Procedure), Section 425, (OECD, 2008). The study was conducted in accordance with Nucro-Technics Study Plan No. CAN/275630, Appendix I.
Justification for Selection of Test System: The test system is internationally accepted for use in acute oral toxicity studies.
Test System:
Test Animal Rattus norvegicus
Strain CD® [Crl:CD®(SD)BR] Sprague-Dawley
Source Charles River Canada, Montreal, Quebec
Number and Sex 5 female rats were used.
Body Weight Range 200.2 - 216.7 g after fasting. The weight
variation in animals at the start of the study
did not exceed ± 20 percent of the mean
weight.
Acclimatization Period 7 days
Age at Study Start Approximately 7 to 8 weeks
Animal Identification Body colour coding, cage labels
Animal Housing and Maintenance
Female rats were individually housed in separate quarters in solid bottom cages. Individual animals were identified by colour coding; the animal number and group number also appeared on the outside of each cage to preclude mix-up. The animal room environment was controlled (targeted ranges: temperature 19 °C to 25 °C, relative humidity 30 - 70%, minimum 10 air changes per hour) and monitored. The photo-cycle was 12 hours light and 12 hours dark. Upon arrival all animals were submitted to a general physical examination and all were found healthy and were admitted. Teklad Certified Rodent Diet and water were offered ad libitum throughout the acclimatization and study periods, except as specified under “Preparation of Animals”. The cage cleaning schedule, air filtration and recirculation, health checks and facility maintenance were carried out in accordance with the applicable Nucro-Technics’ Standard Operating Procedures, and such activities were recorded in the animal room records. Animals were housed and maintained according to the AAALAC International Guide for the Care and Use of Laboratory Animals, CCAC Guidelines for Care and Use of Experimental Animals and Nucro-Technics’ Standard Operating Procedures.
Animal Selection
The test population of animals was randomly selected from newly arrived, previously unused rats.
Preparation of Animals
All animals used for the Limit Test were fasted over-night. Food but not water was withheld beginning at 4:00 p.m. on the day preceding dosing, and was returned to the cages approximately 1 hour after dosing.
Experimental Procedures
Limit Test
The first animal was dosed at 2000 mg/kg. Since this animal survived, four additional animals were sequentially dosed at 2-day intervals. A total of five animals were dosed.
Dose Administration
The test animals were dosed at 2000 mg/kg (2.0 mL/kg by volume of test item). The individual doses of the test item were individually calculated for each animal based on the body weight of the animal. All doses were administered orally using a feeding cannula inserted into the stomach of the animals.
Observations During In-Life Phase
The animals were individually observed once during the first 30 minutes after dosing and periodically during the first 24 hours following dosing (with special attention given during the first 4 hours). Observations once daily were carried out for the remainder of the study. The animals were observed for 14 days after the dosing. Cageside observations were directed towards any changes in the skin and fur; eyes and mucous membranes; respiratory, circulatory, autonomic and central nervous system; and also somatomotor activity and behaviour pattern. Particular attention was directed to any observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and/ or coma. All observations were recorded daily for the entire study period using the In-Life Module V.6.1, and the entries were monitored by the Study Director. The body weights of the animals were determined prior to test item administration (i.e., Day 0), on Day 7 and on Day 14. Body weight gains were calculated. For calculation of LD50, results were entered into the Acute Oral Toxicity statistical program, V.1.0.
Post Mortem Examination
Gross necropsy was performed on each rat at the end of a 14-day observation period and necropsy included an examination of: external surfaces of the body; all orifices; cranial cavity; external surfaces of the brain and spinal cord; nasal cavity and paranasal sinuses; thoracic, abdominal, and pelvic cavities and viscera.
Archive
The original copy of the protocol, and all raw data that were generated at Nucro-Technics and a copy of the final report will be stored in the Nucro-Technics’ archives for 6 years. Nucro-Technics will notify the Sponsor in advance of the end of this period to allow the Sponsor to secure alternative storage facilities, if required.

Statistics:

For calculation of LD50, results were entered into the Acute Oral Toxicity statistical program, V.1.0.

Preliminary study:

The first animal was dosed at 2000 mg/kg. Since this animal survived, four additional animals were sequentially dosed at 2-day intervals. A total of five animals were dosed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortalities were observed post dosing and during the 14-day observation period in any of the animals.

Clinical signs:

other: All clinical signs appear normal

Gross pathology:

No organs with gross findings were observed in necroscopy observations

Other findings:

none

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the foregoing results, the acute oral LD50 in rats of the test item, Tall Oil, Maleated (EnvaMul™ 600) was found to be greater than 2000 mg/kg. Therefore, the test item is considered not to present a significant acute toxic risk if swallowed.

Executive summary:

An Acute Oral Toxicity Study of the test item, Tall Oil, Maleated (EnvaMul™ 600), was carried out at Nucro-Technics according to Study Plan No. CAN/275630. The first animal was dosed at 2000 mg/kg, 2.0 mL/kg by dose volume. Since this first animal survived, four additional test animals were dosed at 2-day intervals. A total of 5 female CD (Sprague-Dawley) rats were dosed. All animals received the test item by oral gavage using a feeding cannula inserted into the stomach of the animals. The animals were observed for a 14-day period after dosing. Body weights were recorded prior to test item administration (i.e., Day 0), on Day 7 and prior to necropsy on Day 14. No mortalities were observed post dosing and during the 14-day observation period in any of the animals. All rats gained body weight by Day 7 and again by Day 14. At the end of a 14-day observation period, each animal was sacrificed and submitted for gross necropsy. No gross pathological findings were observed in any of the rats at necropsy. Based on the foregoing results, the acute oral LD50 in rats of the test item, Tall Oil, Maleated (EnvaMul™ 600), was found to be greater than 2000 mg/kg. Therefore, the test item is considered not to present a significant acute toxic risk if swallowed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

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

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

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:

June-July 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

other: EC No 440/2008, part B: "Acute Toxicity (Dermal)"

Qualifier:

according to guideline

Guideline:

other: JMAFF Guidelines (2000), including the most recent revisions

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Remarks:

Wistar Han

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test System
Species: Rat
Strain: Crl: WI(Han)
Condition: Outbred, SPF-Quality
Source: Charles River Deutschland, Sulzfeld, Germany
Number of Animals: 5 males and 5 females (females were nulliparous and non-pregnant).
Age at the Initiation of Dosing: Young adult animals (approximately 11 weeks old) were selected.
Weight at the Initiation of Dosing: 185 to 295 g.

Justification for Test System and Number of Animals
The Wistar Han rat was chosen as the animal model for this study as recognized by international guidelines as a recommended test system. The test method and number of animals were based on the test guidelines.
This type of study plan was reviewed and agreed by the Laboratory Animal Welfare Officer and the Ethical Committee of Charles River Den Bosch as required by the Dutch Act on Animal Experimentation (February 1997).

Animal Identification
At study assignment, each animal was identified using a tail mark with indelible ink.

Environmental Acclimation
The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

Selection, Assignment, Replacement, and Disposition of Animals
Animals were assigned to the study at the discretion of the coordinating biotechnician according to body weights, with all animals within ±20% of the sex mean. Animals in poor health or at extremes of body weight range were not assigned to the study.
Before the initiation of dosing, a health inspection was performed and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions.
The disposition of all animals was documented in the study records.

Husbandry
Housing
On arrival, animals were group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, animals were individually housed 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. These housing conditions were maintained unless deemed inappropriate by the Study Director and/or Clinical Veterinarian. The room(s) in which the animals were kept were documented in the study records.
Animals were separated during designated procedures/activities. Each cage was clearly labeled.

Environmental Conditions
Target temperatures of 18 to 24 °C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 22 °C with an actual daily mean relative humidity of 48 to 71% (see deviations in Appendix 3). A 12 hour light/12 hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

Food
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility.
It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.

Water
Municipal tap-water was freely available to each animal via water bottles.
Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility.
It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.

Animal Enrichment
For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.

Veterinary Care
Veterinary care was available throughout the course of the study; however, no examinations or treatments were required.

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

A single dose of test item was administered to the appropriate animals by dermal application on Day 1. One day before dosing, an area of approximately 5x7 cm on the back of the animals was clipped. The test item was applied in an area of approximately 10% of the total body surface, i.e. approximately 25 cm² for males and 18 cm² for females. The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

Duration of exposure:

24 h

Doses:

The dose level was 2000 mg/kg body weight.

No. of animals per sex per dose:

5 males and 5 females (females were nulliparous and non-pregnant). Each one exposed to 2000 mg/kg test item

Control animals:

no

Details on study design:

Mortality/Moribundity Checks
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.

Clinical Observations

Postdose Observations
Postdose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days.
All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded (if appropriate), particular attention being paid to the animals during and for the first hour after dosing.

Body Weights
Animals were weighed individually on Day 1 (predose), 8 and 15.

Terminal Procedures
All moribund animals and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Lethargy, hunched posture, rales, piloerection, chromodacryorrhoea (snout), hypothermia and/or hypersensitivity to touch were noted for the animals between Days 1 and 7. Focal erythema, thickened area, alopecia, necrosis, scales, scabs, brown staining we

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

TABLE 1 MORTALITY DATA
TEST DAY HOURS AFTER TREATMENT	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15	'
MALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	.
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	.

TABLE 2 CLINICAL SIGNS
TEST DAY HOURS AFTER TREATMENT	MAX GRADE	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15	'
MALES 2000 MG/KG
ANIMAL 1																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	2	3	2	1	1	1	1	1	-	-	-	-	-	-	.
Necrosis (Treated skin)	(3)	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	-	-	1	1	-	-	-	-	-	-	.
Scabs (Treated skin)	(3)	-	-	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	-	-	.
Various																			.
Hypersensitivity to touch	(1)	-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 2																			.
Behavior																			.
Lethargy	(3)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	-	-	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
Piloerection	(1)	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	-	-	1	1	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	-	-	.
Various																			.
Hypothermia	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 3																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	1	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	1	1	1	1	1	-	-	-	-	-	-	-	-	.
Thickened area (Treated skin)	(3)	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	-	-	-	-	-	-	-	-	.
ANIMAL 4																			.
Behavior																			.
Lethargy	(3)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	1	1	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	2	2	1	1	1	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	2	2	1	1	-	-	-	-	-	-	.
Thickened area (Treated skin)	(3)	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	-	-	.
Secretion / excretion																			.
Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 5																			.
Behavior																			.
Lethargy	(3)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Posture																			.
Hunched posture	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Breathing																			.
Rales	(3)	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	2	2	1	1	1	-	-	-	-	-	-	-	-	.
Piloerection	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	-	1	1	1	-	-	-	-	-	-	.
Scabs (Treated skin)	(3)	-	-	-	-	-	-	-	-	1	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	1	1	1	1	-	-	.
Secretion / excretion																			.
Chromodacryorrhoea (Snout)	(3)	-	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
- = Sign not observed

TABLE 2 CLINICAL SIGNS (continued)
TEST DAY HOURS AFTER TREATMENT	MAX GRADE	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15	'
FEMALES 2000 MG/KG
ANIMAL 6																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	-	-	1	1	1	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	1	2	1	1	1	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	-	1	1	1	-	-	-	-	-	-	.
Thickened area	(3)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
Various																			.
Hypersensitivity to touch	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 7																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	1	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	1	1	1	1	-	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	2	1	1	1	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
Secretion / excretion																			.
Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Various																			.
Hypersensitivity to touch	(1)	-	-	-	1	1	1	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 8																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	1	1	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	1	1	1	1	1	-	-	-	-	-	-	-	-	.
Alopecia	(3)	-	-	-	-	-	-	-	-	1	1	1	1	1	1	1	1	1	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	-	1	1	1	-	-	-	-	-	-	.
Scabs (Treated skin)	(3)	-	-	-	-	-	-	-	-	1	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
ANIMAL 9																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	2	2	2	1	1	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	2	2	1	1	-	-	-	-	-	-	.
Thickened area (Treated skin)	(3)	-	-	-	1	2	1	1	1	-	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
Various																			.
Hypersensitivity to touch	(1)	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	.
ANIMAL 10																			.
Posture																			.
Hunched posture	(1)	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-	-	-	.
Skin / fur																			.
Erythema focal (Treated skin)	(4)	-	-	-	1	2	2	2	1	1	-	-	-	-	-	-	-	-	.
Scales (Treated skin)	(3)	-	-	-	-	-	1	1	2	2	1	1	-	-	-	-	-	-	.
Thickened area	(3)	-	-	-	1	1	1	1	1	-	-	-	-	-	-	-	-	-	.
Brown staining (Treated skin)	(1)	-	-	-	1	1	1	1	1	1	1	1	-	-	-	-	-	-	.
Secretion / excretion																			.
Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	.
Various																			.
Hypersensitivity to touch	(1)	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	.
- = Sign not observed

TABLE 3 BODY WEIGHTS (GRAM)
SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15	'
MALES 2000 MG/KG
	1	295	299	321	.
	2	288	291	317	.
	3	292	302	322	.
	4	282	281	303	.
	5	288	287	315	.
					.
	MEAN	289	292	316	.
	ST.DEV.	5	9	8	.
	N	5	5	5	.
					.
FEMALES 2000 MG/KG
	6	206	208	210	.
	7	210	202	211	.
	8	202	198	203	.
	9	223	226	234	.
	10	185	188	192	.
					.
	MEAN	205	204	210	.
	ST.DEV.	14	14	15	.
	N	5	5	5	.
					.

TABLE 4 MACROSCOPIC FINDINGS
ANIMAL	ORGAN	FINDING	DAY OF DEATH	'
MALES 2000 MG/KG
1		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
2		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
3		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
4		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
5		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
FEMALES 2000 MG/KG
6		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
7		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
8		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
9		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.
10		No  findings noted	Scheduled necropsy	.
			Day 15 after treatment	.

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of EnvaMul 600 in Wistar rats was established to exceed 2000 mg/kg body weight.
Based on these results, EnvaMul 600 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Executive summary:

The objective of this study was to determine the potential toxicity of EnvaMul 600, when given by a single dermal dose.

 

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

·        OECD No. 402 (1987) "Acute Dermal Toxicity"

·        EC No 440/2008, part B: "Acute Toxicity (Dermal)"

·        EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity"

·        JMAFF Guidelines (2000), including the most recent revisions.

 

EnvaMul 600 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

No mortality occurred.

Lethargy, hunched posture, rales, piloerection, chromodacryorrhoea (snout), hypothermia and/or hypersensitivity to touch were noted for the animals between Days 1 and 7.

Focal erythema, thickened area, alopecia, necrosis, scales, scabs, brown staining were seen in the treated skin-area of the animals during the observation period. These local effects were considered not to have affected the conclusion of the study.

The mean body weight gain during the observation period was within the range expected for rats used in this type of study.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The dermal LD50 value of EnvaMul 600 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, EnvaMul 600 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

The substance was tested for acute oral and dermal toxicity to rats up to limit dose of 2000 mg/kg bw. No mortality was observed. Therefore, the substance is not subject to classification for acute toxicity by oral or dermal exposure, according to CLP (Regulation EC No 1272/2008).