Poly(oxy-1,2-ethanediyl), α-[2-(dide- cylmethylammonio)ethyl]- .omega.- hydroxy-, propanoate (salt) (Bardap 26)

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2 April 2001 to 4 May 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

bis(decyl)(2-hydroxyethyl)methylazanium propanoate

EC Number:

619-057-3

Cas Number:

94667-33-1

Molecular formula:

C29 H62 N O4 . C3 H5 O2

IUPAC Name:

bis(decyl)(2-hydroxyethyl)methylazanium propanoate

Constituent 2

Reference substance name:

alpha.-[2-(Didecylmethylammonio)ethyl]-.omega.-hydroxy-poly(oxy-1,2-ethanediyl) propionate

IUPAC Name:

alpha.-[2-(Didecylmethylammonio)ethyl]-.omega.-hydroxy-poly(oxy-1,2-ethanediyl) propionate

Constituent 3

Reference substance name:

N,N-Didecyl-N-methyl-poly(oxyethyl)ammonium Propionate

IUPAC Name:

N,N-Didecyl-N-methyl-poly(oxyethyl)ammonium Propionate

Constituent 4

Reference substance name:

Bardap 26

IUPAC Name:

Bardap 26

Details on test material:

The test substance was stored at room temperature and expected to be stable for the duration of testing (expiry date 12-12-2001).

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Male and female Sprague-Dawley rats were obtained from Ace Animals, Inc., Boyertown, Pennsylvania, USA, aged 8-12 weeks and weighing 204-234 g (males) and 150-184g (females). The animals were housed singly in suspended stainless steel cages, the temperature and relative humidity were 18-23°C and 35-72%, respectively, lighting was provided according to a 12 hour light/dark cycle. The rats were provided with food (Purina Rodent Chow #5012) ad libitum, except during fasting prior to dosing (approximately 18 hours pre-dose and 3.5 hours after dosing). Water (filtered tap water)was provided ad libitum. Rats in the range finding screen were acclimatised for 29 days; the test animals were acclimated for 10 days.
In-life dates of main test animals: 20 April to 4 May, 2001.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Individual doses were calculated based on initial body weights, taking into account the specific gravity of the test substance. Each rat received the appropriate amount of test substance as a single administration by gavage.

Doses:

500, 1000, 2000, 4000 mg/kg bw.

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

A range finding screen was conducted prior to the main test for selection of dose levels, with four male and four female rats. The test substance was administered to fasted rats at 500 and 2000 mg/kg bw. The animals were observed for 7 days after dosing. One male and one female from the 2000 mg/kg bw group died (50% mortality). Based on the range finding, dose levels of 500, 1000, 2000 and 4000 mg/kg bw were selected for the main test.
The observation period was 14 days. The rats were observed for clinical signs and mortality, and body weights were recorded. Gross necropsy was performed on rats that died during the study and on survivors at the end of the 14 day observation period.

Statistics:

Moving Average Method was used to calculate the LD50.

Results and discussion

Preliminary study:

Range-finding: one male and one female from the 2000 mg/kg bw group died (50% mortality).

Effect levels

Effect levels 1

Sex:

male

Dose descriptor:

LD50

Effect level:

972 mg/kg bw

Based on:

test mat.

95% CL:

>= 798 - <= 1 237

Effect levels 2

Sex:

female

Dose descriptor:

LD50

Effect level:

1 414 mg/kg bw

Based on:

test mat.

95% CL:

>= 845 - <= 2 367

Effect levels 3

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

1 157 mg/kg bw

Based on:

test mat.

95% CL:

>= 833 - <= 1 519

Mortality:

There were no mortalities at 500 mg/kg bw. There was a dose-related increase in mortality in the remaining groups:
1000 mg/kg bw: 4/10 (3/5 males; 1/5 females), 2000 mg/kg bw: 9/10 (5/5 males; 4/5 females), and 4000 mg/kg bw: 10/10 (5/5 males; 5/5 females). Mortalities in the 1000 and 2000 mg/kg bw groups occurred within 7 days of administration, and within 2 days of administration in the 4000 mg/kg/ bw group.

Clinical signs:

other: 500 mg/kg bw: most rats exhibited ocular discharge, hunched posture, hypoactivity, ano-genital staining, soft faeces, diarrhoea and/or reduced faecal volume. All animals recovered by Day 10. 1000 mg/kg bw: clinical signs in the 3 males and 1 female that

Gross pathology:

No gross abnormalities were observed in the animals that survived to study termination. Necropsy of the animals that died following administration of 1000 mg/kg bw revealed discolouration of the lungs, liver and intestines and/or gaseous distention of the intestines. Necropsy of the animals that died following administration of 2000 mg/kg bw revealed discolouration of the lungs, liver and/or intestines, and in one male the stomach was filled with fluid and the lungs were oedematous. Necropsy of the 4000 mg/kg bw group revealed discolouration of the lungs, liver and/or intestines, oedema of the lungs, fluid-filled stomach, gaseous distention of the intestines and/or rigor mortis.

Other findings:

No other findings were reported.

Any other information on results incl. tables

Table 1. Group incidence of mortality

Dose level (mg/kg bw)	Mortality^#
	Male	Female	Total
500	0/5	0/5	0/10
1000	3/5	1/5	4/10
2000	5/5	4/5	9/10
4000	5/5	5/5	10/10

^# Number dead/total number in group

Table 2. Group incidence of clinical signs (number affected/total number in group)

Clinical signs	Dose level (mg/kg bw)
	500	1000	2000	4000
Ocular discharge	1/10	2/10	1/10	0/10
Hunched/abnormal posture	1/10	2/10	7/10	3/10
Hypoactivity	1/10	5/10	10/10	10/10
Ano-genital staining	6/10	7/10	9/10	5/10
Soft faeces	2/10	4/10	2/10	0/10
Diarrhoea	1/10	3/10	9/10	5/10
Reduced faecal volume	7/10	10/10	5/10	3/10
Facial staining	0/10	1/10	2/10	0/10
Irregular/shallow breathing	0/10	1/10	4/10	6/10
Piloerection	0/10	1/10	2/10	1/10
Distended abdomen	0/10	1/10	0/10	0/10
Prone	0/10	0/10	1/10	6/10

 Table 3. Group mean body weight (g)

Day of study	Dose level (mg a.s./kg)
	500		1000		2000		4000
	M	F	M	F	M	F	M	F
0	215	175	221	176	219	165	213	170
7	235	209	235	187	a/d	178	a/d	a/d
14	294	244	305	221	a/d	211	a/d	a/d

M: male

F: female

a/d: all dead

Table 4. Group incidence of gross findings at necropsy

Gross findings^#	Dose level (mg a.s./kg)
	500	1000	2000	4000
No abnormalities	10/10	6/10	1/10	0/10
Lungs
Slightly red	0/10	0/10	2/10	2/10
Slightly red, oedematous	0/10	0/10	1/10	4/10
Moderately red	0/10	4/10	7/10	4/10
Liver
Discolouration	0/10	4/10	5/10	8/10
Intestines
Slightly red	0/10	0/10	1/10	4/10
Slightly red, gaseous distention	0/10	2/10	0/10	0/10
Red, gaseous distention	0/10	0/10	0/10	4/10
Red	0/10	0/10	5/10	0/10
Red/green	0/10	2/10	0/10	0/10
Black/green/red	0/10	0/10	3/10	0/10
Stomach
Filled with fluid	0/10	0/10	1/10	6/10
General appearance
Rigor mortis	0/10	0/10	0/10	4/10

^# Gross abnormalities seen only in decedents
Number of animals affected/total number in group

Applicant's summary and conclusion

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The combined sexes LD50 is 1157 mg/kg bw. The test substance is therefore classified as H302: Harmful if Swallowed according to Regulation (EC) No 1272/2008.

Executive summary:

The acute oral toxicity of Bardap 26 was determined in male and female Sprague-Dawley rats. The test substance was administered by gavage to groups of 5 rats/sex at doses of 500, 1000, 2000 and 4000 mg/kg bw. The rats were observed for clinical signs, mortality and body weights changes for 14 days after administration. Gross necropsy was performed on rats that survived to study termination and those that died during the study.

There was a dose-related increase in mortality; 3/5 males and 1/5 females in the 1000 mg/kg bw groups died within 7 days of administration; 5/5 males and 4/5 females in the 2000 mg/kg bw group died within 7 days of administration; all animals (5 males and 5 females) in the 4000 mg/kg bw group died within 2 days of administration. Clinical signs included hypoactivity, irregular/shallow breathing, ano-genital staining and diarrhoea. Gross necropsy findings in decedents included discoloured liver, red lungs, black/green/red intestines and fluid-filled stomach. The LD₅₀ for male rats was 972 mg/kg (95% confidence limits: 798-1237 mg/kg bw); the LD₅₀ for female rats was 1414 mg/kg bw (95% confidence limits: 845-2367 mg/kg bw); and the LD₅₀ for both sexes combined was 1157 mg/kg bw (95% confidence limits: 833-1519 mg/kg bw). The substance is therefore classified as H302: Harmful if Swallowed according to Regulation (EC) No 1272/2008.