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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 January 1985 - 2 July 1985
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed similar to OECD 425 with some additional investigations. The study was not performed under GLP but in-house quality assurance was in place.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report date:
1985

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
GLP compliance:
no
Remarks:
in-house QA was in place
Test type:
up-and-down procedure
Limit test:
no

Test material

Constituent 1
Reference substance name:
Triamine C16-18, C18-unsaturated, branched
Cas Number:
1219826-66-0
IUPAC Name:
Triamine C16-18, C18-unsaturated, branched
Constituent 2
Reference substance name:
Amines, N-(3-aminopropyl)-N-tallow alkyltrimethylenedi-
EC Number:
288-048-0
EC Name:
Amines, N-(3-aminopropyl)-N-tallow alkyltrimethylenedi-
IUPAC Name:
288-048-0
Details on test material:
Test substance name: Lilamin LS 33, batch: 5/83
Chemical name: a modified tallow alkyl dipropylene triamine
Appearance: light brown/yellow liquid
Storage: ambient temperature
Relative density: 0.86
Technical grade; equivalent to Lilamin LSP 33.
60-65% tallow alkyl dipropylene triamine
25-30% tallow diamine
5-10% ditallow alkyl propylene diamine

Test animals

Species:
rat
Strain:
other: CFY (Sprague-Dawley Origin)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Interfauna UK Ltd., (formerly Hacking and Churchill Limited,) England
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 91-150g
- Fasting period before study: overnight
- Housing: They were housed in groups by sex or single in metal cages with wire mesh floors.
- Diet (e.g. ad libitum): ad libitum, standard laboratory rodent diet (Labsure LAD 1)
libitum. The batch
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22
- Humidity (%): 56
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 22 January 1985 To: 2 July 1985

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: unchanged or in distilled water
Details on oral exposure:
Administered as supplied or as 25% w/v in distilled water.
VEHICLE
- Concentration in vehicle: 25% w/v
- Amount of vehicle (if gavage): 0.1 ml/kg
- Justification for choice of vehicle: no data
- Lot/batch no. (if required): no data
- Purity: no data

MAXIMUM DOSE VOLUME APPLIED: unchanged: 2.3 ml/kg in distilled water: 0.1 ml/kg
Doses:
Stage I: 2000, 200, 25 mg/kg bw.
Stage II: males, 640, 800, 1000, 1260 mg/kg bw
females, 320, 400, 500, 640, 800, 1000 mg/kg bw

Stage III:
320, 200, 80 mg/kg bw
No. of animals per sex per dose:
Stage I: 2
Stage II: One animal per dosing occasion.
Stage III: 5
Control animals:
other: Not in stage I and II but at stage III a control group was included.
Details on study design:
Stage I, II and III
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed soon after dosing; then at frequent intervals for the remainder of Day 1. On
subsequent days the animals were observed at least twice per day. Clinical signs were recorded at each observation. Individual bodyweights of rats on Days 1 (day of dosing), 4, 8 and 15 and at death.
- Necropsy of survivors performed: yes
- Other examinations performed: other: only in stage III
On Days 2 and 7 samples of blood were withdrawn, under light ether anaesthesia, from the orbital sinus of all rats. The blood samples were placed in EDTA anti-coagulant tubes for subsequent; haernatological investigations. The following estimations were performed:
Total white cell count (WBC Total), Differential WBC counts - standard, microscopy of blood smear, stained with modified Wright's stain counting 100 cells, Neutrophils (N), Lymphocytes (L), Eosinophils (E), Basophils (B), Monocytes (M).
Statistics:
The LD50 and its 95% confidence limits were estimated for both males and females separately, based on the method of Brownlee, Hodges and
Rosenblatt. For haematological data statistical analysis was carried out using the individual animal as the basic experimental unit. The following sequence of statistical tests was used for haematological parameters in which the relative frequency of the mode was less than 75% (i.e. the data did not consist predominant of one particular value):

(i) Bartlett's test was applied to test for heterogeneity of variance between treatments. Where significant (at the 1% level) heterogeneity was found, a log transformation was tried to see if the heterogeneity was removed.
(ii) If no satisfactory transformation was found the Kruskal-Wallis analysis of ranks was used.
(iii) Analyses of variance were followed by Student's 't' test for a-dose-related response. The Kruskal-Wallis analyses were followed by the non-parametric equivalent of the, ' t' test.

For those parameters in which the relative frequency of the mode is 75% or more, the number of animals in each group with values different
from the mode will be analysed using Fisher's exact test and Mantel's test. Fisher's test will be used to detect general differences between treatments, whereas Mantel's test will be used specifically for detecting dose-related trends in the numbers of such animals.

Results and discussion

Effect levelsopen allclose all
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 200 - < 2 000 mg/kg bw
Remarks on result:
other: Stage I
Sex:
male
Dose descriptor:
LD50
Effect level:
895 mg/kg bw
95% CL:
657 - 1 219
Remarks on result:
other: Stage II
Sex:
female
Dose descriptor:
LD50
Effect level:
637 mg/kg bw
95% CL:
467 - 869
Remarks on result:
other: Stage II
Sex:
male/female
Dose descriptor:
other: Haematological changes
Remarks on result:
other: No effects observed after a single dose of 80 mg/kg bw on day 2 and day 7 after treatment.
Mortality:
Stage I:
All rats dosed at 2000 mg/kg were found dead within one or two days of treatment.

Stage II:
Death occurred amongst male rats at 800 mg/kg and above and amongst female rats at 500 mg/kg and above within one to four days of dosing.

Stage III:
Based on the results from stage I and II rats were dosed at the expected maximum sub-lethal dose.
The treatmnet started with 320 mg/kg bw but one female rat was found dead within one day. A further group of ten rats was therefore dosed at a lower level of 200 mg/kg. Two male rats were found dead within. one day of dosing. A third group of ten rats was then dosed at 80 mg/kg and no mortalities occured.
Clinical signs:
other: Stage I: Signs of reaction to treatment observed shortly after dosing in all rats were pilo-erection, hunched posture and abnormal gait (waddling). These signs were accompanied by: increased salivation in all rats at 200 and 2000 mg/kg, pallor of extremit
Gross pathology:
Stage I:
In the animals that died at 2000 mg/kg bw autopsy revealed congestion or haemorrhage of the lungs and pallor o f the liver, spleen and kidneys. In the surviving animals no abnormalities were observed.

Stage II:
Autopsy of the animals that died revealed congestion of the lungs and pallor of the liver, spleen and kidneys. Pallor of the kidney cortex was also observed in one male rat at 1000 mg/kg. In the surviving animals no abnormalities were observed.

Stage III:
No abnormalities were observed.

Other findings:
Haematology:
No statistically significant effects were noted.

Any other information on results incl. tables

Stage I:

Time and number of deaths:

 sex  Dose mg/kg bw  No. of deaths  Day 1  2 a  2b  3a  3b  4-15
 m  25  0            
 m  200  0            
 m  2000  2    1    1    
 f 25   0            
 f  200  0            
 f  2000  2    1  1      

a = first observation, b = second observation, m = male, f = female

Stage II:

Mortality data:

                             mortality data on each dosing occasion
 sex  dose (mg/kg)  1
 m  640                0  
 m  800  0        0    1*    
 m  1000    0    1*    1*      
m  1260      1*            
 f  320        0          
 f  400          0        
 f  500      1*      0      
 f  640    1*          0    
 f  800  1              0  
 f  1000                  1*

1 = death, 0 = survival, * = late death

Time and number of deaths:

       Day
 Sex  Dose (mg/kg)  No. death/No. dosed  1 2a   b 3a  b 4 a  b  5a  6 -15
 m  640  0/1                    
 m  800  1/3        1            
 m  1000  2/3        1        1    
 1260  1/1        1            
 f  320  0/1                    
 f  400  0/1                    
 f  500  1/2      1              
 f  640  1/2        1            
 f  800 1/2     1                
 f  1000  1/1        1            

a = first observation, b = second observation, m = male, f = female

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The LD50 in rats was calculated to be 766 mg/kg bw for males and females combined.
Executive summary:

An acute oral toxicity study was performed with "Lilamin LS 33". Animals were administered the test substance by oral gavage. The study was divided in three stages. Stage on was a range finding study were rats (2/sex dose) were given a single dose of 2000, 200 or 25 mg/kg bw. Based on these results an up-and-down procedure was followed dosing 1 animal at a time to determine a more exact LD50. In phase III, based on the LD50 findings, 10 animals (5/sex) were given a dose of 80 mg/kg bw and haematological parameters were measured 2 and 7 days after dosing. The LD50 in rats is between 200 and 2000 mg/kg bw, 895 (CI 657-1219) mg/kg bw for males and 637 (CI 467-869) mg/kg bw for females. Based on the performed acute oral toxicity studies the test substance should be classified as harmful (Xn and R22). No effects on haematology were observed after a single oral dose of 80 mg/kg bw.