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Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2016

Materials and methods

Test guidelineopen allclose all
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

Test material

1
Chemical structure
Reference substance name:
Heptanal, oligomeric reaction products with aniline
EC Number:
500-007-3
EC Name:
Heptanal, oligomeric reaction products with aniline
Cas Number:
9003-50-3
Molecular formula:
(C7H14O)n.(C6H7N)n
IUPAC Name:
Heptanal, oligomeric reaction products with aniline
Test material form:
liquid
Details on test material:
Test item: 207368/A
Identification: Hepteen Base®
Appearance: Clear amber liquid (determined by Charles River Den Bosch)
Batch: LT5C30Y170
Purity/Composition: 99.7%
Test item storage: At room temperature
Stable under storage conditions until: 30 November 2016 (retest date)
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

Test animals

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.

Administration / exposure

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).

Results and discussion

Preliminary study:
No preliminary study conducted
Effect levels
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

Any other information on results incl. tables

 

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

 

 

Applicant's summary and conclusion

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.