Trimethyl[2-(phosphonooxy)ethyl]ammonium chloride, calcium salt (1:1)

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 10, 2017 - May 24, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

calcium tetrahydrate chloride [2-(trimethylazaniumyl)ethyl] phosphate

Cas Number:

72556-74-2

Molecular formula:

C5H21CaClNO8P

IUPAC Name:

calcium tetrahydrate chloride [2-(trimethylazaniumyl)ethyl] phosphate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SPF Caw

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: JANVIER LABS (53940 Legenest St. Isle - France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: males: 7 weeks old; females: 8 weeks old
- Weight at study initiation: males: 257.4 ± 9.1; females: 214.2 ± 5.7
- 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): 19ºC - 25ºC. A temperature lower than 19ºC was registered on 18 of May 2017. The minimum value measured was 27ºC.
- Humidity (%): 30% - 70%. A relative humidity lower than 30% was registered on 10, 13, 16, 17, 23 and 24 of May 2017. The minimum value measured was 18%.
- 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: 10 May 2017 To: 24 May 2017

Administration / exposure

Type of coverage:

other: non-occlusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk of the animal
- % coverage: 10% of the body surfade area
- Type of wrap if used: non-occlusive porous gauze dressing (50 mm x 50 mm non woven swab, 4-layer patch, MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic micropore TM adhesive tape from 3M)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): after removing the gauze dressings, the treated area was rinsed with destilled water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Concentration (if solution): 0.2 g/ml
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 mL/Kg bw

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

yes

Remarks:

Study No. TAD-2016-004 (see "Other information on results").

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily observations, the animals were weighed on day 0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes.
At the end of the study, the animals were euthanized with sodium pentobarbital (Dolethal®). Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.
Gross pathology: On D14, the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. 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, Ileon, 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, Treatment site, Mortality.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

Neither cutaneous reactions nor systemic clinical signs related to the administration of the test item were observed.

Body weight:

The body weight evolution of the animals remained normal throughout the study.

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.

MALES	D0	D2			D2-D0	D7	D7-D0		D14	D14-D0
Rm 1455	261	266			5	310	49		354	93
Rm 1456	255	261			6	313	58		360	105
Rm 1457	258	262			4	311	53		348	90
Rm 1458	244	256			12	298	54		344	100
Rm 1459	269	280			11	318	49		369	100
MEAN	257.4	265.0			7.6	310.0	52.6		355.0	97.6
Standard deviation	9.1		9.1	3.6		7.4	3.8	9.9			6.0
FEMALES	D0		D2	D2-D0		D7	D7-D0	D14			D14-D0
Rf 1460	211		216	5		231	20	300			89
Rf1461	220		218	-2		238	18	269			49
Rf 1462	218		219	1		240	22	264			46
Rf 1463	206		212	6		216	10	248			42
Rf 1464	216		228	12		239	23	256			40
MEAN	214.2		218.6	4.4		232.8	18.6	267.4			53.2
Standard deviation	5.7		5.9	5.3		10.0	5.2	19.9			20.3

GENERAL APPEARANCE BEFORE AUTOPSY: Normal

Observations: Nothing to report.

Current control Study:

The study identified No. TAD -2016 -004 was performed to assess the comportament of the strain of rat used at the laboratory in the environment and to give additional historical data. The method was designed to meet the requirements of the following: OECD Guideline No. 402 (February 24th, 1987) and Method B.3 of Council regulation No. 440/2008.

 

Test item : Distilled water

Study dates: 20 September 2016 to 04 October 2016

Results:

- Clinical examinations: Nothing to report. Animal normal (10/10)

- Bodyweight evolution: normal during the test.

- Necropsy: No treatment related changes were observed.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 of the test item in rats is greater than 2000 mg/kg bw.

Executive summary:

A limit test was performed to determine the acute dermal toxicity of the test item in rats according to OECD 402 and EU method B.3, following GLP. The test item was applied onto the intact skin of 10 Sprague Dawley rats (5 males, 5 females) at a dose of 2000 mg/kg bw. All animals were observed once daily for 14 days, survival and body weight were monitored. There were no mortality and systemic clinical signs related to the administration of the test item during the study. Neither cutaneous reactions nor systemic clinical signs related to the administration of the test item were observed. Normal changes in body weights and none macroscopic findings were found. In conclusion, the test item was found to be non toxic by dermal route, with an LD50 > 2000 mg/kg bw in rats.