Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 Aug - 19 Sep 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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:

8 Feb 2002

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: EPA OPPTS 870.1000 (Acute toxicity testing background)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates

EC Number:

306-227-4

EC Name:

Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates

Cas Number:

96690-34-5

Molecular formula:

C4H11O4P to C8H20O7P2 as representative molecular formula of the composition as specified in section 1.2

IUPAC Name:

Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 8 - 11 weeks
- Weight at study initiation: 144 - 154 g (animals 1-3; step 1), 170 - 178 g (animals 4-6; step 2), 150 - 165 g (animals 7-9; step 3) and 152 - 160 g (animals 10-12; step 3)
- Fasting period before study: 16 to 19 h (access to water was permitted)
- Housing: animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 030512).
- Diet: Altromin 1324 maintenance diet for rats and mice (lot no. 0939), ad libitum
- Water: tap water, sulphur acidified to a pH value of approximately 2.8, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: aqua ad injectionem (sterile water)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/mL (2000 mg/kg bw dose groups) and 0.03 g/mL (300 mg/kg bw dose groups)
- Amount of vehicle: 10 mL/kg bw
- Justification for choice of vehicle: the vehicle was chosen due to its non-toxic characteristics.
- Lot/batch no.: 10952-1

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: For all animals of step 1 and step 2, 1.5 g of the test item were emulsified with the vehicle to gain a final volume of 7.5 mL and to achieve a dose of 2000 mg/kg body weight at a dose volume of 10 mL/kg body weight. For all animals of step 3 and step 4, 0.3 g of the test item were emulsified with the vehicle to gain a final volume of 10 mL and to achieve a dose of 300 mg/kg body weight at a dose volume of 10 mL/kg body weight. The dose formulations were made shortly before each dosing occasion. Homogeneity of the test item in the vehicle was maintained by vortexing the prepared suspension for 10 min before every dose administration.

Doses:

Step 1 and 2: 2000 mg/kg bw
Step 3 and 4: 300 mg/kg bw

No. of animals per sex per dose:

3 per step

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were weighed on Day 1 (prior to the administration) and on Days 8 and 15. A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). Thereafter, animals were observed for clinical signs once daily until the end of the 14-day observation period.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

500 mg/kg bw

Based on:

test mat.

Mortality:

Step 1 (2000 mg/kg bw): 1/3 females died
Step 2 (2000 mg/kg bw): 3/3 females died
Step 3 and 4 (300 mg/kg bw): 0/3 females died

Clinical signs:

other: 2000 mg/kg bw: reduced spontaneous activity, moving the bedding, kyphosis, bradykinesia, piloerection, tremor and salivation; in the surviving animals of this dose group, all symptoms recovered within up to 2 days post dose. 300 mg/kg bw: reduced spontane

Gross pathology:

Necropsy revealed no substance-related findings in surviving animals. Macroscopic findings of the four animals found dead at 2000 mg/kg bw involved bleeding nose (1/4 dead animals) and a bloated stomach (4/4 dead animals) containing remains of the test item (3/4 dead animals).

Any other information on results incl. tables

Table 1. Clinical signs and mortality - acute oral toxicity

Step	Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death (dead animals/3 animals)	Mortality (%)
1	2000	1/3/3	5 min - Day 2	2 h 10 min	33
2	2000	3/3/3	10 min - death	1 h 40 min (1/3) + Day 2 (2/3)	100
3	300	0/3/3	5 min - Day 2	---	0
4	300	0/3/3	10 - 30 min	---	0
LD50 cut-off = 500 mg/kg bw

  * first number = number of dead animals, second number = number of animals with clinical signs, third number = number of animals used

         

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

CLP: Acute Oral 4, H302