Reaction mass of isopentyl salicylate and 2-methylbutyl salicylate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 9th, 2018 to May 29th, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction Mass of Isoamyl salicylate and 2-Methylbutyl salicylate

IUPAC Name:

Reaction Mass of Isoamyl salicylate and 2-Methylbutyl salicylate

Test material form:

liquid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: JANVIER LABS (53940 Legenest St. Isle - France)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-9 weeks old
- Weight at study initiation: 213 ± 16.3 g
- Fasting period before study: overnight.
- Housing: Groups of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air-conditioned animal husbandry.
- Diet (e.g. ad libitum): Foodstuff (ENVIGO -2016) ad libitum.
- Water (e.g. ad libitum): Tap-water from public distribution system ad libitum. Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas- Eurofins (France).
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 ºC
- Humidity (%): 30-70%
- Air changes (per hr): 10 changes/hour
- Photoperiod (hrs dark / hrs light): 12 h light (7 - 19 h) / 12 h darkness

IN-LIFE DATES: From March 7th, 2018 To May 23rd, 2018

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.39 mL/kg (test item), 10 mL/kg (control).
CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: no indication of toxicity based on available data. Therefore, the selected starting dose was 2000 mg/kg body weight.

Doses:

2000 mg/kg

No. of animals per sex per dose:

6

Control animals:

yes

Remarks:

Study No. TAO423-2018-001 (see "Other information on results").

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Systemic observations at 30 min, 1h, 3h, 4h, 24h, 48h
after administration and daily for 14 days. Weighing: on day 0 (just before administering the test item) then on Day 2, Day 7, and Day 14.
- Necropsy of survivors performed: yes. At termination, macroscopic observations were performed. Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. Parameters examined: Oesophagus, Stomach, Duodenum, Jejunum, Ileum, Caecum, Colon, Rectum, Spleen, Liver, Thymus, Trachea, Lungs, Heart, Kidneys, Urinary Bladder, Ovaries, Uterus, Treatment Area, Adrenals and Pancreas.
- Other examinations performed: Clinical signs: Spontaneous activity, Preyer's reflex (noise), Respiratory rate, Convulsions, Tremors, Body temperature, Muscle tone, Palpebral opening, Pupil appearance, Salivation, Lachrymation, Righting reflex, Back hair appearance, Mortality.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: A decrease of spontaneous activity (6/6), muscle tone (5/6) and righting reflex (5/6) was noted a t30 minutes post dose. The animals recovered normal activity at 24 hours post dose.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal treatment-related changes.

Any other information on results incl. tables

Table 1. Test item at 2000 mg/kg bw. Body weight and weight gain in grams.

FEMALES	D0	D2	D2-D0	D7	D7-D0	D14	D14-D0
Rf 2488 Rf 2489 Rf 2490	191 200 196	225 220 210	24 20 14	238 249 247	47 49 51	250 270 265	59 70 69
Rf 2532 Rf 2533 Rf 2534	224 236 211	243 250 228	19 14 17	256 267 238	32 31 27	271 282 249	47 46 38
MEAN	209.7	229.3	19.7	249.2	39.5	264.5	54.8
Standard deviation	17.5	14.8	7.4	11.1	10.6	12.9	13.2

Table 2. Clinical signs

OBSERVATIONS:	FEMALES			FEMALES
T0 + 30 minutes	Rf 2488	Rf 2489	Rf 2490	Rf 2532	Rf 2533	Rf 2534
Spontaneous activity	D	D	D	D	D	D
Preyer’s reflex (noise)	N	N	N	N	N	N
Respiratory rate	N	N	N	N	N	N
Convulsions	N	N	N	N	N	N
Tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	D	D	D	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	N	N	N	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Righting reflex	D	D	D	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

OBSERVATIONS:	FEMALES			FEMALES
T0 + 1 hour	Rf 2488	Rf 2489	Rf 2490	Rf 2532	Rf 2533	Rf 2534
Spontaneous activity	D	D	D	N	D	D
Preyer’s reflex (noise)	N	N	N	N	N	N
Respiratory rate	N	N	N	N	N	N
Convulsions	N	N	N	N	N	N
Tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	N	N	N	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Righting reflex	N	N	N	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

OBSERVATIONS:	FEMALES			FEMALES
T0 + 3 hours T0 + 4 hours	Rf 2488	Rf 2489	Rf 2490	Rf 2532	Rf 2533	Rf2534
Spontaneous activity	D	D	D	N	D	D
Preyer’s reflex (noise)	N	N	N	N	N	N
Respiratory rate	N	N	N	N	N	N
Convulsions	N	N	N	N	N	N
Tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	D	D
Palpebral opening	N	N	N	N	N	N
Pupil appearance	N	N	N	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Righting reflex	N	N	N	N	D	D
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

OBSERVATIONS:	FEMALES			FEMALES
D1 to D14	Rf 2488	Rf 2489	Rf 2490	Rf 2532	Rf 2533	Rf 2534
Spontaneous activity	N	N	N	N	N	N
Preyer’s reflex (noise)	N	N	N	N	N	N
Respiratory rate	N	N	N	N	N	N
Convulsions	N	N	N	N	N	N
Tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	N	N	N	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Righting reflex	N	N	N	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

Table 3. Necropsy findings

Found dead:  GENERAL APPEARANCE		Euthanasia BEFORE AUTOPSY:Normal		X	At	term	X
			Observed Organs	Observations
* SOPHAGUS * STOMACH * DUODENUM * JEJUNUM * ILEON * CAECUM * COLON * RECTUM * SPLEEN * LIVER			X X X X X X X X XX	N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R.
* THYMUS			X	N.t.R.
* TRACHEA * LUNGS * HEART			XXX	N.t.R. N.t.R. N.t.R.
* KIDNEYS * URINARYBLADDER * OVARIES * UTERUS			X X X X	N.t.R. N.t.R. N.t.R. N.t.R.
* TREATMENT AREA			-	-
* ADRENALS * PANCREAS			XX	N.t.R. N.t.R.
PARTICULARS: None

Found dead:  GENERAL APPEARANCE		Euthanasia: BEFORE AUTOPSY:Normal		X	At	term	X
			Observed Organs	Observations
* ESOPHAGUS * STOMACH * DUODENUM * JEJUNUM * ILEON * CAECUM * COLON * RECTUM * SPLEEN * LIVER			X X X X X X X X XX	N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R. N.t.R.
* THYMUS			X	N.t.R.
* TRACHEA * LUNGS * HEART			X X X	N.t.R. N.t.R. N.t.R.
* KIDNEYS * URINARYBLADDER * OVARIES * UTERUS			X X X X	N.t.R. N.t.R. N.t.R. N.t.R.
* TREATMENT AREA			-	-
* ADRENALS * PANCREAS			XX	N.t.R. N.t.R.
PARTICULARS: None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

EU Criteria

Conclusions:

The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD 423 and EU B.1 tris, under GLP conditions. A limit test was performed, where the test item was administered to a total of 6 female Sprague-Dawley rats (class method) at the dose of 2000 mg/kg body weight by oral gavage. All animals were observed immediately after administration for the onset of any toxic signs and once daily thereafter for 14 days. No mortality occurred during the study and the body weight evolution of the animals remained normal during the study. The decrease of spontaneous activity (6/6), muscle tone (5/6) and righting reflex (5/6) was noted a t30 minutes post dose. The animals recovered normal activity at 24 hours post dose. Macroscopic examination of the animals at the end of the study did not reveal treatment related changes. No other signs of systemic toxicity were noted. The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in the rat.