Sodium long chain alkyl salicylate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 August 2014 to 09 September 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF, 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., sodium salts

Molecular formula:

Na(C21-25H33-41O3)

IUPAC Name:

Benzoic acid, 2-hydroxy-, mono-C14-18-alkyl derivs., sodium salts

Test material form:

other: sticky solid

Details on test material:

- Appearance: Green-black solid
- Storage conditions of test material: At room temperature in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Strain: Wistar strain Crl:WI (Han) (outbred, SPF-Quality)
- Age at study initiation: Approximately 8 to 9 weeks old
- Weight at study initiation: 147 to 169 g. Body weight variation did not exceed ± 20 % of the sex mean.
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3 to 4 hours after administration of the test material. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labelled cages (height 18 cm) containing sterilised sawdust as bedding material and paper as cage-enrichment.
- Diet: Free access to pelleted rodent diet
- Water: Free access to tap water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 to 24 °C
- Humidity: 40 to 70 % (relative)
- Air changes: at least 10 air changes/hour
- Photoperiod: 12-hour light/12-hour dark cycle

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE:
- Specific gravity: 1.036
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at the testing facility and on test material data.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bodyweight

DOSAGE PREPARATION (if unusual): The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. The concentration of the test material in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
In order to obtain homogeneity, the test material formulations were heated in a water bath with a maximum temperature of 77.5 °C for a maximum of 75 minutes. The test material formulations were allowed to cool down to a temperature of maximally 40 °C prior to dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Starting dose = 2000 mg/kg bw. Substance initially assumed to be of low toxicity based on test on amixture containing the substance.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 animals were dosed at 2000 mg/kg bw. 6 animals were dosed at 300 mg/kg bw.

Control animals:

no

Details on study design:

- Study design: The toxicity of the test material 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. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups. Initially, the test material was administered at 2000 mg/kg bw. In a stepwise procedure, two additional groups were dosed at 300 mg/kg bw.
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Animals were observed for mortality and viability twice daily. Time of death was recorded as precisely as possible. Body weights were recorded on Day 1 (pre-administration of the test material), Days 8 and 15 and at death (if found dead after Day 1). At periodic intervals on the day of dosing (Day 1) and once daily thereafter until Day 15, animals were observed for clinical signs. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded.
- Necropsy of survivors performed: Yes. At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy.
- Other examinations performed: Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

No statistical analysis was performed as the method used is not intended to allow the calculation of a precise LD50 value.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

300 - 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, two animals were found dead on day 2.
At 300 mg/kg, no mortality occurred.

Clinical signs:

other: At 2000 mg/kg, hunched posture, piloerection and/or diarrhoea were noted for the animals on Day 1. At 300 mg/kg, hunched posture, piloerection and/or diarrhoea were noted for the animals on Days 1 and 2.

Gross pathology:

At 2000 mg/kg, abnormalities of the stomach (glandular mucosa: several dark red foci; forestomach: irregular surface) and thymus (reddish discolouration) were found in both animals that died during the study. Additionally, in one of these animals gelatinous contents of the gastro intestinal-tract were found.
Furthermore, advanced autolysis was seen in both animals found dead. This finding was considered to be due to the time elapsed between death and macroscopic examination.
At 300 mg/kg, no abnormalities were found at macroscopic examination of the animals.

Other findings:

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 1000 mg/kg body weight.

Any other information on results incl. tables

Table 1: Body Weight Data (g)

Dose Group	Animal Number	Day 1	Day 8	Day 15
Group dosed with 2000 mg/kg bw	1	147*	-	-
	2	162	183	202
	3	164**	-	-
	Mean (SD)	158 (9)	183	202
First group dosed with 300 mg/kg bw	4	153	178	206
	5	153	190	197
	6	155	191	206
	Mean (SD)	154 (1)	186 (7)	203 (5)
Second group dosed with 300 mg/kg bw	7	169	203	213
	8	159	187	198
	9	161	180	194
	Mean (SD)	163 (5)	190 (12)	202 (10)

*Animal found dead on day 2. Body weight at death 139 g

**Animal found dead on day 2. Body weight at death 160 g

Applicant's summary and conclusion

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study the LD50 value was established to be within the range of 300 to 2000 mg/kg body weight and in accordance with OECD Guideline 423, the LD50 cut-off value was considered to be 1000 mg/kg.

Executive summary:

The potential of the test material to cause acute oral toxicity in the rat was investigated in accordance with the standardised guidelines OECD 423, EU Method B.1 tris, EPA OPPTS 870.1100 and JMAFF 12 Nousan, Notification No 8147 under GLP conditions.

Initially, the test material was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure, two additional groups of three females were dosed at 300 mg/kg body weight. All 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).

At 2000 mg/kg, two animals were found dead on day 2. At 300 mg/kg, no mortality occurred.

At 2000 mg/kg, hunched posture, piloerection and/or diarrhoea were noted for the animals on Day 1. At 300 mg/kg, hunched posture, piloerection and/or diarrhoea were noted for the animals on Days 1 and 2.

The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

At 2000 mg/kg, abnormalities of the stomach (glandular mucosa: several dark red foci; forestomach: irregular surface) and thymus (reddish discolouration) were found in both animals that died during the study, at macroscopic post mortem examination. Additionally, in one of these animals gelatinous contents of the gastro intestinal-tract were found. At 300 mg/kg, no abnormalities were found at macroscopic examination of the animals.

Under the conditions of this study the LD50 value was established to be within the range of 300 to 2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 1000 mg/kg body weight.