Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute Toxicity via Dermal Route

Key value determined in a GLP accredited laboratory study using the standard acute method, in accordance with OECD Guideline 402, EU Method B.3, JMAFF, 12 Nousan, Notification No 8147 and EPA OPPTS 870.1200.

The dermal LD50 value of Hepteen Base ® in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute Toxicity via Oral Route

Key value determined in a GLP accredited laboratory study using the standard acute method, in accordance with OECD Guideline No. 423 (2001), EU method B1, US EPA OPPTS 870.1100 (2002) and JMAFF Guidelines (2000).

The oral LD50 of Hepteen Base ® in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute Toxicity via Inhalation Route

Endpoint waived. Substance supplied liquid form and does not have the propensity to produce airborne dust. In addition physicochemical data on the test substance showed that it had a low vapour pressure and no boiling point, which indicates the liquid is not volatile consequently  route of exposure to humans or the environment via the inhalation route would not be possible REACH REGULATION Annex VII COLUMN 2: SPECIFIC RULES FOR ADAPTATION FROM COLUMN 1 - 7.14.  "The study does not need to be conducted if the substance is marketed or used in a non solid or granular form." Test data has been provided for dermal and oral Acute Toxicity endpoints see sections 7.2.1 and 7.2.3 for more details.

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:
18 April 2016 to 10 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Study was conducted according to OECD, EU, US EPA and JMAFF test guidelines in an accredited GLP laboratory
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001

Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred. Day 10 Group 2, no observations for clinical signs performed
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
May 2008, including themost recent amendments.
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred. Day 10 Group 2, no observations for clinical signs performed.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
EPA 712-C-02-190, 2002.
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred. Day 10 Group 2, no observations for clinical signs performed.
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan,Notification No 8147, November 2000; including the most recent partial revisions.
Deviations:
yes
Remarks:
Deviations from the minimum level of daily mean relative humidity occurred. Day 10 Group 2, no observations for clinical signs performed.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
pH (1% in water, indicative range) 8.0 – 7.8 (determined by Charles River Den Bosch)
Specific gravity/density 0.9130
Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han)
Sex:
female
Details on test animals or test system and environmental conditions:
Species: Rat, Wistar strain Crl:WI (Han) (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals: 6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals.
Age and body weight: Young adult animals (approx. 8-9 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Earmark and tail mark
Health inspection At least prior to dosing. It was ensured that the animals were healthy and without any abnormality that might have affected the study integrity.

Animal Husbandry
Conditions
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle: the photoperiod was between 07:00 and 19:00 hrs daily. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

Accommodation
Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
Acclimatization period was at least 5 days before start of treatment under laboratory conditions.

Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Water
Free access to tap water.
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Method: Oral gavage, using plastic feeding tubes. The test item was stirred on a magnetic stirrer during dosing.
Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item. Water was available ad libitum.
Doses:
Single dose of 2000 mg/kg (2.19 mL/kg) body weight on Day 1.
No. of animals per sex per dose:
2 consecutive groups of 3 female rats at 2000 mg/kg body weight
Control animals:
no
Details on study design:
The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.
The second group of animals at 2000 mg/kg was dosed one week after the first group.
Statistics:
The LD50 cut-off value was established based on OECD guideline 423. No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
Preliminary study:
No preliminary study conducted
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal was found dead on Day 9. No further mortality occurred.
Clinical signs:
Hunched posture, piloerection and/or ptosis were noted for the animals between Days 1 and 5.
Body weight:
Body weight loss or reduced body weight gain was noted for all animals during the first week of the study. The surviving animals gained weight between Days 1 and 15.
Gross pathology:
Abnormalities of the stomach (forestomach: irregular surface) and small intestines (wall: dark red discouloration) were noted for the animal that was found dead during the study, at macroscopic post mortem examination. Macroscopic examination of the other animals did not reveal any abnormalities.
Other findings:
None

 

Mortality Data

Test day

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Hours after treatment

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Females 2000 mg/kg

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Females 2000 mg/kg

-

-

-

-

-

-

-

-

-

-

1

-

-

-

-

-

-

 

-         = no mortality

Clinical signs

Test day

Max grade

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Hours after treatment

 

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Females 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

1

1

1

1

1

1

1

-

-

-

-

-

-

-

-

-

-

Skin/fur Piloerection

(1)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Various Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Animal 2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

1

1

1

1

1

1

-

-

-

-

-

-

-

-

-

-

-

Skin/fur Piloerection

(1)

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Various Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Animal 3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

1

1

1

1

1

1

-

-

-

-

-

-

-

-

-

-

-

Skin/fur Piloerection

(1)

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Various Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Animal 4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

-

1

-

-

-

-

-

-

-

-

 

 

 

 

 

 

 

Skin/fur Piloerection

(1)

-

1

-

-

-

-

-

-

-

-

 

 

 

 

 

 

 

Various Ptosis

(3)

-

1

-

-

-

-

-

-

-

-

 

 

 

 

 

 

 

Animal 5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur Piloerection

(1)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Various Ptosis

(3)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Animal 6

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

(1)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur Piloerection

(1)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Various Ptosis

(3)

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-         = sign not observed

Body weight (grams)

Sex/dose level

Animal

Day 1

Day 8

Day 15

FEMALES 2000 MG/KG

1

159

58

194

 

2

138

132

157

 

3

158

132

172

 

Mean

152

141

174

 

ST. Dev.

12

15

19

 

N

3

3

3

FEMALES 2000 MG/KG

 

 

 

 

 

4

152

125*

---

 

5

172

168

186

 

6

161

161

194

 

Mean

162

151

190

 

ST. Dev.

10

23

6

 

N

3

3

2

* Animal was found dead on Day 9. Body weight at death: 115 gram

 

Macroscopic findings

Animal

Organ

Finding

Day of death

FEMALES 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

FEMALES 2000 MG/KG

 

 

 

4

Stomach

Small intestines

Forestomach: irregular surface,

Wall: discolouration, dark red.

Spontaneous death Day 9 after treatment

5

 

No findings noted

Scheduled necropsy Day 15 after treatment

6

 

No findings noted

Scheduled necropsy Day 15 after treatment

 

 

Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2500 mg/kg body weight. Based on these results:

-according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), Hepteen Base® does not have to be classified and has no obligatory labelling requirement for oral toxicity.
Executive summary:

Assessment of acute oral toxicity with Hepteen Base® in the rat (Acute Toxic Class Method).

 

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

OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method"

Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method" EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

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

 

Hepteen Base® was administered by oral gavage to two consecutive groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

 

One animal was found dead on Day 9. No further mortality occurred.

 

Hunched posture, piloerection and/or ptosis were noted for the animals between Days 1 and 5.

 

Body weight loss or reduced body weight gain was noted for all animals during the first week of the study. The surviving animals gained weight between Days 1 and 15.

 

Abnormalities of the stomach (forestomach: irregular surface) and small intestines (wall: dark red discouloration) were noted for the animal that was found dead during the study, at macroscopic post mortem examination.

Macroscopic examination of the other animals did not reveal any abnormalities.

 

The oral LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2500 mg/kg body weight. Based on these results:

 

-according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), Hepteen Base® does not have to be classified and has no obligatory labelling requirement for oral toxicity.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
K1 - Study performed in GLP accredited laboratory to recognised OECD, EU & EPA standards.

Acute toxicity: via inhalation route

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:
18 April 2016 to 03 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Study was conducted in accordance with OECD, EU, EPA and Japanese test guidelines, in compliance with GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Species: Rat, Wistar strain, Crl:WI (Han) (outbred, SPF-Quality).
Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source: Charles River Deutschland, Sulzfeld, Germany.
Number of animals: 5 males and 5 females (females were nulliparous and non-pregnant).
Age and body weight: Young adult animals (approx. 10 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification: Tail mark with indelible ink.
Health inspection: At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality.

Animal Husbandry
Conditions
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle: the photoperiod was between 07:00 and 19:00 hrs daily. The light/dark cycle may be interrupted for study related activities. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

Accommodation
Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).

Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Water
Free access to tap water.
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Single dosage, on Day 1.
Duration of exposure:
24 hours, after which dressings were removed and the skin cleaned of residual test item using tap water.
Doses:
2000 mg/kg (2.19 mL/kg) body weight.
No. of animals per sex per dose:
males and 5 females (females were nulliparous and non-pregnant).
Control animals:
no
Details on study design:
Test Item Preparation
The test item was dosed undiluted as delivered by the Sponsor.
No correction was made for the purity/composition of the test item.

Treatment
Method Dermal application. The test item was stirred on a magnetic stirrer during application.
Clipping One day before exposure (Day -1) an area of approximately 5x7 cm on the back of each animal was clipped.
Application: The test item was applied on an area of approx. 10% of the total body surface, i.e. approx. 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.
*. Manufacturers: Laboratoires Stella s.a., Liege, Belgium (surgical gauze) and 3M, St. Paul, Minnesota, U.S.A. (Coban & Micropore).
Frequency: Single dosage, on Day 1.
Dose level (volume): 2000 mg/kg (2.19 mL/kg) body weight.
Application period: 24 hours, after which dressings were removed and the skin cleaned of residual test item using tap water.

Observations
Mortality/Viability Twice daily.
Body weights Days 1 (pre-administration), 8 and 15.
Clinical signs At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
Necropsy At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Evaluation
A dermal LD50 value was derived.
The results were evaluated according to:
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments);
- 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).
Statistics:
Not specified.
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:
Hunched posture, piloerection, quick breathing, shallow respiration, chromodacryorrhoea and/or ptosis were noted for the animals between Days 1 and 3

General erythema, erythema maculate, scales, scabs, wound and/or thickened areas were noted for the skin of the animals during the observation period. These local effects were considered not to have affected the conclusion of the study.
Body weight:
Body weight loss or reduced body weight gain was noted for the male animals between Days 1 and 8.

The changes noted in body weight gain in females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
None specified

Mortality Data

Test day

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Hours after treatment

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Males 2000 MG/KG

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Females 2000 MG/KG

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

 

Clinical signs

Test day

 

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Hours after treatment

Max grade

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Males 2000 MG/KG

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

ANIMAL 1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

2

2

1

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

1

1

Scabs

(3)

-

-

-

-

-

1

1

1

1

1

1

1

1

-

-

-

-

Scabs (Flank left)

(3)

-

-

-

-

-

-

-

-

-

-

-

-

-

1

1

1

1

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (Snout)

(3)

-

2

2

1

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema

(4)

-

-

-

3

3

2

2

-

-

-

-

-

-

-

-

-

-

Erythema maculate (treated skin)

(4)

-

-

-

2

2

2

2

1

1

1

1

1

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scabs (Left flank)

(3)

-

-

-

-

-

-

-

-

-

-

-

-

-

1

1

1

1

Scabs (Treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Secrretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (snout)

(3)

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Ptosis

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

 

 

 

1

 

 

 

 

 

 

 

 

 

 

 

 

 

Shallow respiration

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General Erythema (Treated skin)

(4)

-

-

-

2

2

2

2

2

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

-

-

Scabs (Flank left)

(3)

-

-

-

-

-

-

-

-

-

-

-

-

-

1

1

1

1

Scabs (treated skin)

(3)

-

-

-

-

-

-

-

-

-

-

1

1

1

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (Snout)

(3)

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

1

1

-

-

-

-

-

-

-

-

-

-

Erythema maculate (treated skin)

(4)

-

-

-

3

3

2

2

2

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

1

1

Scabs (Flank left)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Scabs (Treated skin)

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (eye left)

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Chromodacryorrhoea (eye right)

(3)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Chromodacryorrhoea (Snout)

(3)

-

2

2

2

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

1

1

1

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scales (treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

1

1

Scabs (treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (snout)

(3)

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Ptosis

(3)

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

FEMALES 2000 MG/KG

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

ANIMAL 6

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

2

2

2

1

-

-

-

-

-

-

-

-

Erythema maculate (Treated skin)

(4)

-

-

-

1

2

1

1

-

-

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

-

-

-

-

Scabs (Treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (snout)

(3)

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 7

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

1

1

1

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

-

-

Scabs (Treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Thickened are (Treated area)

(3)

-

-

-

1

1

1

1

-

-

-

-

-

-

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (snout)

(3)

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Various

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Ptosis

(3)

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 8

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (treated skin)

(4)

-

-

-

1

2

1

1

-

-

-

-

-

-

-

-

-

-

Erythema maculate (Treated skin)

(4)

-

-

-

1

2

1

1

-

-

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

-

-

-

-

Scabs(Treated skin)

(3)

-

-

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (snout)

(3)

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

ANIMAL 9

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

2

2

1

1

-

-

-

-

-

-

-

-

-

-

Erythema maculate (Treated skin

(4)

-

-

-

2

2

1

1

-

-

-

-

-

-

-

-

-

-

Scales (Treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

1

1

-

-

Scabs (Treated skin)

(3)

-

-

-

-

1

1

1

1

1

1

1

1

1

-

-

-

-

Thickened are (Treated skin)

(3)

-

-

-

1

1

1

1

-

-

-

-

-

-

-

-

-

-

ANIMAL 10

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Posture

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Hunched posture

(1)

-

-

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Breathing

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Quick breathing

(1)

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/fur

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

General erythema (Treated skin)

(4)

-

-

-

3

2

2

2

-

-

-

-

-

-

-

-

-

-

Erythema Maculate (Treated skin)

(4)

-

-

-

2

2

1

1

2

1

-

-

-

-

-

-

-

-

Piloerection

(1)

-

-

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

Scales (treated skin)

(3)

-

-

-

-

-

-

-

1

1

1

1

1

1

-

-

-

-

Thickened area (Treated skin)

(3)

-

-

-

1

1

1

1

-

-

-

-

-

-

-

-

-

-

Secretion/excretion

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Chromodacryorrhoea (Snout)

(3)

-

1

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

- = Sign not observed


 

Body weights

SEX/DOSE LEVEL

Animal

Day 1

Day 8

Day 15

Males 2000 MG/KG

 

 

 

 

 

1

325

331

353

 

2

342

329

359

 

3

332

323

359

 

4

341

338

36

 

5

325

319

345

 

Mean

333

328

356

 

ST.DEV.

8

7

8

 

N

5

5

5

Females 2000 MG/KG

 

 

 

 

 

6

193

198

209

 

7

201

207

209

 

8

193

199

205

 

9

197

207

219

 

10

196

205

206

 

Mean

196

203

210

 

ST.DEV.

3

4

6

 

N

5

5

5

 

Macroscopic findings

Animal

Organ

Finding

Day of death

Males 2000 MG/KG

 

No findings noted

Scheduled necropsy Day 15 after treatment

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 Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, Hepteen Base® 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:

Assessment of acute dermal toxicity with Hepteen Base® in the rat. The study was carried out based on the guidelines described in:

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

-Commission Regulation (EC) No 440/2008, B3: "Acute Toxicity (Dermal)"

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

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

Hepteen Base® 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.

Hunched posture, piloerection, quick breathing, shallow respiration, chromodacryorrhoea and/or ptosis were noted for the animals between Days 1 and 3

Body weight loss or reduced body weight gain was noted for the male animals between Days 1 and 8.

The changes noted in body weight gain in females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

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

The dermal LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, Hepteen Base® 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
Quality of whole database:
K1 - Study performed in GLP accredited laboratory to recognised OECD, EU & EPA standards.

Additional information

Acute Toxicity via Dermal Route

Hepteen Base® 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.

Hunched posture, piloerection, quick breathing, shallow respiration, chromodacryorrhoea and/or ptosis were noted for the animals between Days 1 and 3

Body weight loss or reduced body weight gain was noted for the male animals between Days 1 and 8.

The changes noted in body weight gain in females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

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

The dermal LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, Hepteen Base® 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).

Acute Toxicity via Oral Route

Hepteen Base® was administered by oral gavage to two consecutive groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily

observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

 

One animal was found dead on Day 9. No further mortality occurred.

 

Hunched posture, piloerection and/or ptosis were noted for the animals between Days 1 and 5.

 

Body weight loss or reduced body weight gain was noted for all animals during the first week of the study. The surviving animals gained weight between Days 1 and 15.

 

Abnormalities of the stomach (forestomach: irregular surface) and small intestines (wall: dark red discouloration) were noted for the animal that was found dead during the study, at macroscopic post mortem examination.

Macroscopic examination of the other animals did not reveal any abnormalities.

 

The oral LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2500 mg/kg body weight. Based on these results:

 

-according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), Hepteen Base® does not have to be classified and has no obligatory labelling requirement for oral toxicity.

Acute Toxicity via Inhalation Route

Endpoint waived. Substance supplied liquid form and does not have the propensity to produce airborne dust. In addition physicochemical data on the test substance showed that it had a low vapour pressure and no boiling point, which indicates the liquid is not volatile consequently  route of exposure to humans or the environment via the inhalation route would not be possible REACH REGULATION Annex VII COLUMN 2: SPECIFIC RULES FOR ADAPTATION FROM COLUMN 1 - 7.14.  "The study does not need to be conducted if the substance is marketed or used in a non solid or granular form." Test data has been provided for dermal and oral Acute Toxicity endpoints see sections 7.2.1 and 7.2.3 for more details.

Justification for classification or non-classification

Acute Toxicity via Dermal Route

According to the OECD 402 test guideline, the dermal LD50 value of Hepteen Base® in Wistar rats was established to exceed 2000 mg/kg body weight. Based on these results, Hepteen Base® 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).

Acute Toxicity via Oral Route

According to the OECD 423 test guideline, the oral LD50 cut-off value was considered to be 2500 mg/kg body weight for Hepteen Base ®. Based on these results, Hepteen Base ® does not have to be classified and has no obligatory labelling requirements for acute oral toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments). It does however meet the classification criteria for Acute Oral Toxicity Category 5 and as such is obligated to adhere to the labelling requirements under the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2016) (including all amendments).

Acute Toxicity via Inhalation Route

Endpoint waived. Substance supplied liquid form and does not have the propensity to produce airborne dust. In addition physicochemical data on the test substance showed that it had a low vapour pressure and no boiling point, which indicates the liquid is not volatile consequently  route of exposure to humans or the environment via the inhalation route would not be possible REACH REGULATION Annex VII COLUMN 2: SPECIFIC RULES FOR ADAPTATION FROM COLUMN 1 - 7.14.  "The study does not need to be conducted if the substance is marketed or used in a non solid or granular form." Test data has been provided for dermal and oral Acute Toxicity endpoints see sections 7.2.1 and 7.2.3 for more details.