Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2011-05-31 to 2011-06-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The reliability is rated 1 because the study followed the standard guideline of reference (OECD 423), which describes a procedure designed to evaluate this endpoint. The results were reviewed for reliability and assessed as valid. The study was conducted under GLP condition.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report date:
2011

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
tetrakis{9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethylxanthylium} C12-(branched and/or linear)-alkyl-(4-sulfonatophenoxy)benzenesulfonate and oxybis[C12-(branched and/or linear)-alkyl-benzenesulfonate]
EC Number:
700-761-5
Molecular formula:
not applicable
IUPAC Name:
tetrakis{9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethylxanthylium} C12-(branched and/or linear)-alkyl-(4-sulfonatophenoxy)benzenesulfonate and oxybis[C12-(branched and/or linear)-alkyl-benzenesulfonate]
Details on test material:
- Name of test material (as cited in study report): Sepisol Fast Red SN
- Substance type: organic salt - UVCB
- Physical state: Red powder
- Lot/batch No.: 029244
- Storage condition of test material: Room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Elevage JANVIER (53940 Le Genest St Isle - France).
- Age at study initiation: From 8 to 9 weeks.
- Weight at study initiation: Between 194 g and 219 g.
- Fasting period before study: Food was removed one day before the beginning of the study and redistributed 4 hrs after the test item administration.
- Housing: Housed by group of 3 in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid, containing sawdust bedding which was changed at least twice per week.
- Diet : Foodstuff (M20-SDS) ad libitum.
- Water : Tap water ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C
- Humidity (%):30 to 70 %
- Air changes (per hr): Approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2g (for the first step), 0.3 g (for the second and third step) of the test item was weighed in a 10 mL volumetric flask and olive oil was added. The preparation was magnetically stirred to obtain a homogeneous pink suspension, just before being administered by gavage using a suitable syringe graduated fitted with an oesophageal metal canula.

Doses:
2000, 300 and 50 mg/kg
No. of animals per sex per dose:
3 female/dose/step
Step 1: 2000 mg/kg b.w.
Step 2: 300 mg/kg b.w.
Step 3: 300 mg/kg b.w.
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily examination. Weighed on days D0, D2, D7 and D14.
- Necropsy of survivors performed: yes
- Other examinations performed: Observation and mortality report carried out every day : examination of behavioural or toxic effects on the major physiological functions:
* Body weight
* Clinical signs (spontaneous activity, Preyer's reflex, respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, mydriasis, salivation, lachrymation, righting reflex, piloerection, mortality)
* Autopsy

Results and discussion

Mortality:
Step 1 at 2000 mg/kg : Death of the 3 rats at 22 hrs 45 min post-dose (2/3) and 47 hrs 45 min post-dose (1/3)
Step 2 at 300 mg/kg: Death of 1 rat on D3 post-dose (1/3)
Step 3 at 300 mg/kg: No death (0/3)
Clinical signs:
At 2000 mg/kg (3 rats treated): Pink diarrhoea (3/3), pink urine (1/3) and piloerection (3/3).

At 300 mg/kg (6 rats treated): Decrease in spontaneous activity, pink diarrhoea, pink urine and piloerection in the dead rat. No clinical signs in all the survivors
Body weight:
At 2000 mg/kg: A decrease in the body weight of the animals: between -9.5% and -11% compared to day 0.
At 300 mg/kg: A decrease in the body weight in the animal found dead: -22% the day of the death to day 0.
Gross pathology:
At 2000 mg/kg (all dead): The macrosopical examination of the dead animals revealed black spots on the corpus (2/3), pink content or pink coloration of the stomach and intestines (3/3), red coloration of the lungs (2/3), thinning of the forestomach (1/3).

At 300 mg/kg: The macroscopical examination of the dead animal (1/6) revealed a thinning of the forestomach, a pink coloration of stomach and intestines and a red coloration of the lungs.
Other findings:
Before the macroscopical examination,
- at 2000 mg/kg, a rigor mortis (1/3) was noted.

Any other information on results incl. tables

Control study TAO-2011 -002 on the Olive oil used as the vehicule in this study

The study was performed from 2011 -03 -29 to 2011 -04 -12 to assess the comportment of the strain of rat used at Phycher laboratory in its environment and to give additional historical data.

The method was designed to meet the requirements of the following:

- OECD guideline for the testing of chemicals N°423

- Method B.1tris of the council regulation N°440/2008

Three animals received the Olive oil, administered by gavage under a volume of 10 mL/kg body weight using a suitable syringe graduated fitted with an oesophageal metal canula.

No clinical signs, body weight changes nor treatment related changes were reported (3/3 animals normal)

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 of the test item Sepisol Fast Red SN is higher than 300 mg/kg and lower than 2000 mg/kg b.w. by oral route in rats.
In accordance with the OECD guideline No. 423, the LD50 cut-off of the test item may be considered as 500 mg/kg body weight by oral route in the rat.
In accordance with the regulation EC No. 1272/2008 (CLP), the test item must be classified in category 4. The signal word "Warning" and hazard statement H302 "Harmful if swallowed" are required.
Executive summary:

The test item Sepisol Fast Red SN was administered to a group of 3 females Sprague Dawley rats at the single dose of 2000 mg/kg b.w., then to 2 groups of 3 female Sprague Dawley rats at the single dose of 300 mg/kg b.w.

The experimental protocol was established according to the official method as defined in the OECD guideline No. 423 and the test method B.A tris of the Council regulation No. 440/2008.

It was noted the death of the three rats treated at 2000 mg/kg b.w. during the first step of the study (3/3) at 22 hours 45 minutes post-dose (2/3) and at 47 hours 45 minutes post-dose (1/3). The mortalities were preceded by pink diarrhoea (3/3), pink urine (1/3) and piloerection (3/3).

A decrease in body weight of the animals was also noted: between -9.5% and -11% on the day of the death, compared to day 0.

Before the macroscopical examination, a rigor mortis was noted in the animal found dead at 47 hours 45 minutes post-dose. The macroscopical examination of the dead animals revealed black spots on the corpus (2/3), pink contents or pink coloration of the stomach and intestines (3/3), red coloration of the lungs (2/3), thinning of the forestomach (1/3).

It was noted the death of one rat treated at 300 mg/kg b.w. during the second step of the study (1/6) on day 3. The mortality was preceded by decrease in spontaneaous activity, pink diarrhoea, pink urine and piloerection.

A decrease in body weight of the animal was also noted: -22% on the day of the death, compared to day 0.

The macroscopical examination of the dead animal revealed thinning of the forestomach, pink coloration of stomach and intestines, red coloration of the lungs.

In the surviving animals treated at 300 mg/kg b.w; (5/6), no clinical signs related to the administration of the test item were observed. The body weight evolution of the animals remained normal throughout the study.

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

 

In conclusion, the LD50 of the test item Sepisol Fast Red SN is higher than 300 mg/kg and lower than 2000 mg/kg body weight by oral route in rat. In accordance with the OECD guideline N° 423, the LD50 cut-off of the test item may be considered as 500 mg/kg body weight by oral route in the rat.

 

According to the criteria for classification, packaging and labeling of dangerous substances and preparations in accordance with the E.E.C. Directives 67/548, 2001/59 and 99/45, the test item Sepisol Fast Red SN must be classified R22 “if swallowed”. The item must be characterised by the symbol “Xn” and the warning label “Harmful”.

In accordance with the Regulation EC N° 1272/2008 (CLP), the test item must be classified in category 4. The signal word “Warming” and hazard statement H 302 “Harmful if swallowed” are required.