Tall oil, compd. with triethanolamine

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Embryotoxicity of Sixteen Industrial Amines to the Chicken Embryo

Author:

Korhonen, A., Hemminki, K., and Vainio, H.

Year:

1983

Bibliographic source:

Institute of Occupational Health, Department of Industrial Hygiene and Toxicology, Haartmaninkatu 1, SF-002090 Helsinki 39, Finland.

Materials and methods

Test guideline

Guideline:

other: Not specified

Principles of method if other than guideline:

The embryotoxic effects of TEA were tested in the chicken embryo assay. White Leghorn chicken eggs (3-days old) were used for the study. Acetone (p.a. grade) was used as the solvent for Triethanolamine (TEA, technical grade). The solution was injected into the egg in a total volume of 5 µL. Ten control eggs, injected with 5 µL of acetone, were incubated with each batch of eggs. Results from solvent controls were not subtracted from experimental values. The background level of 1-2% was insignificant against the variation of the method.

Three-day (72-76 h) embryos were selected by candling. The method used involved the injection on the inner shell membrane, focusing the solution drop exactly on the embryo. After two days post injection, the eggs were candled again. Eggs containing dead embryos were counted and discarded, the remaining eggs were then candled every second or third day. Those containing dead embryos were opened and checked for external malformations and the developmental stage according to V. Hamburger and H. L. Hamilton, A series of normal stages in the development of the chick embryo. J. Morphol. 88, 49-92 (1951 ).

Incubation terminated was 11 days post the injection, after a total incubation of 14 days. The remaining eggs were opened and the embryos inspected for survival and for external malformations. The temperature was kept at 37.7 “C and the humidity between 66% and 71% throughout the incubation period. The eggs were turned two-four times per day.

The affected embryos were classified into the following categories:

(1) Early deaths, embryos that died before day 5 of incubation, within two days of treatment.
(2) Late deaths, non-malformed, externally normal embryos that died between days 5 and 14.
(3) Late deaths, malformed, externally malformed embryos that died between days 5 and 14.
(4) Malformed survivors, externally malformed embryos alive on day 14 of incubation.

LD50 and ED50 values were calculated according to the method described by A. P. Rosiello, J. M. Essigmann and G. N. Wogan, Rapid and accurate determination of the median lethal dose (LD,,) and its error with a small computer. J. Toxicol. Environ. Heaffh 3, 8. K. S. Khera, Ethylenethiourea. Teratogenicity study in rats 797-809 (1977). using a Wang table computer.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

2,2',2''-nitrilotriethanol

EC Number:

203-049-8

EC Name:

2,2',2''-nitrilotriethanol

Cas Number:

102-71-6

Molecular formula:

C6H15NO3

IUPAC Name:

2,2',2''-nitrilotriethanol

Test material form:

not specified

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Merck, Darmstadt. Lot/batch number not specified
- Expiration date of the lot/batch: Not specified
- Purity test date: Not specified (p.a grade)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not specified
- Stability under test conditions: Not specified
- Solubility and stability of the test substance in the solvent/vehicle: Not specified
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Not specified

Test animals

Species:

other: Chicken

Strain:

other: White Leghorn

Remarks:

Test conducted on White Leghorn chicken embryos within the egg

Details on test animals or test system and environmental conditions:

- CHICKEN EMBRYOS
- Source: Hatchery of Siipikarjanhoitajain Liitto ry, Hãmeenlinna, Finland
- Age at study initiation: 3 day embryos

ENVIRONMENTAL CONDITIONS
- Incubator: Commercial type incubator (Manufactured by Hãmeen Insinööritoimisto Oy, Hãmeenlinna.
- Temperature (°C): 37.7 “C
- Humidity (%): 66 - 71 %
- Photoperiod (hrs dark / hrs light): Not specified

IN-LIFE DATES: 3 day old embryos with incubation terminated on day 11 (total incubation period 14 days)

Administration / exposure

Route of administration:

other: Injection into egg

Vehicle:

acetone

Remarks:

p.a. grade

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Treatment concentrations of 1.3, 2.6, 5.2 and 10.5 µmol per egg TEA in acetone injected into the egg in a total volume of 5 µL/per egg.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Not specified, publication details doses soluble or suspendable in acetone in 5 ul of acetone
- Concentration in vehicle: Not specified
- Lot/batch no. (if required): Not specified
- Purity: Not specified (p.a. grade)

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

11 days

Frequency of treatment:

Single treatment in 3 day old embryos which were exposed to TEA in acetone for 11 days

Duration of test:

14 days

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

1.3 other: µmol

Remarks:

1.3 µmol per egg

Doses / concentrations 2

Dose / conc.:

2.6 other: µmol

Remarks:

2.6 µmol per egg

Doses / concentrations 3

Dose / conc.:

5.2 other: µmol

Remarks:

5.2 µmol per egg

Doses / concentrations 4

Dose / conc.:

10.5 other: µmol

Remarks:

10.5 µmol per egg

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Not specified
- Rationale for animal assignment (if not random): Not specified.
- Other: Three days embryos were selected by candling to undergo injection of test item. Two days after the injection with TEA in acetone (as solvent), the eggs were candled again. Eggs containing dead embryos were counted and discarded. The remaining eggs were then candled every second or third day. Those containing dead
embryos were opened and the embryos checked for external malformations and the developmental stage according to Hamburger and Hamilton. The incubation was terminated 11 days after the injection, after a total incubation of 14 days. The remaining eggs were opened and the embryos inspected for survival and for external malformations.

Results and discussion

Results: maternal animals

Effect levels (maternal animals)

Dose descriptor:

other:

Remarks:

Not relevant for this type of study

Effect level:

> 0 - < 0 mg/kg bw/day (nominal)

Based on:

other: Not relevant

Remarks:

Not relevant for this type of test

Basis for effect level:

other: Not applicable for this type of test

Remarks on result:

not measured/tested

Results (fetuses)

Details on embryotoxic / teratogenic effects:

No effects observed with the lowest dose of 1.3 µmol per egg.

At 2.6 µmol per egg, 40% of embryos were observed to have had early deaths (before day 5); 3% had a late death/were non-malformed embryos; and 3% were observed to be malformed embryos at days 6-14. Total was 47% of all affected embryos at this dose.

At 5.2 µmol per egg, 80% of embryos were observed to have had early deaths (before day 5); 3% had a late death/were non-malformed embryos; 3% were observed to be malformed embryos at days 6-14; and 3% were observed to be malformed survivors on day 14. Total was 90% of all affected embryos at this dose.

At the maximum dose of 10.5 µmol per egg, 95% of embryos were observed to have had early deaths (before day 5). Total was 95% of all affected embryos at this dose.

Effect levels (fetuses)

Dose descriptor:

other:

Remarks:

Not applicable for this type of toxicity test

Effect level:

> 0 - < 0 mg/kg bw/day (nominal)

Based on:

other: Not relevant for this type of test

Remarks:

Not applicable tot his type of toxicity test

Basis for effect level:

other: Not applicable

Remarks on result:

not measured/tested

Remarks:

Not applicable for this type of toxicity test

Overall developmental toxicity

Key result

Developmental effects observed:

yes

Lowest effective dose / conc.:

2.6 other: µmol per egg

Treatment related:

yes

Relation to maternal toxicity:

not specified

Dose response relationship:

yes

Relevant for humans:

not specified

Any other information on results incl. tables

Table 1. Embryotoxic effects of triethanolamine on White Leghorn chicken embryos

Treatment (µmol per egg)       Treated embryos (n)    Early deaths (before day 5)   Late deaths non-malformed embryos     (Days 6 -14) malformed embryos     Malformed survivors     All affected embryos     (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  1.3  30  0  0  0  0  0  0  0  0  0  0  2.6  30  12  40  1  3  1  3  0  0  14  47  5.2  30  24  80  1  3  1  3  1  3  27  90  10.5  20  19  95  0  0  0  0  0  0  19  95

Table 2. ED50 and LD50 values on different embryotoxic effects of Triethanolamine on White Leghorn chicken embryos

Total effect    ED50  (µmol per egg) 95% confidence limits      Total mortality     Early deaths (LD50)     (µmol per egg)   Tan α   (µmol per egg)   Tan  α (µmol per egg)   Tan α   2.6  (1.9 - 3.5)  1.6  2.6  1.6  3.0  1.5

Table 3. Malformation types produced in White Leghorn chicken embryos by Triethanolamine

Small head  Small eye cup     Defects of lids and cornea     Defects of beak     Encephalocoele or skin pimple in head     Open coelom     Short back or neck     Defects of wings     Defects of legs       Oedema and lymph blebs    Number of malformed embryos  Number    of malformations   (n) (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  (n)  (%)  -  -  -  -  -  -  -  -  -  - 3  100  1  33  -  -  -  -  1  33     3   5

Applicant's summary and conclusion

Conclusions:

The embryotoxic effects of TEA were tested in the chicken embryo assay to evaluate embryotoxicity as part of the evaluation of 16 industrial chemicals. Three-day-old white Leghorn chicken eggs were injected with 1.3-10.5 µmol of TEA in acetone. Eleven days after the injection (total incubation period 14 days), eggs were opened and the embryos inspected for survival and external malformations. The ED50 for embryotoxic effects was 2.6 µmol/egg. The embryotoxic effects included early mortality and malformations (open coelom, short back or neck, edema, and lymph blebs). The incidence of malformations (3%-6%) in the TEA-treated groups was not significantly different from that of controls.

Executive summary:

Triethanolamine (TEA, p.a grade) was tested for embryotoxic effects in the chicken embryo assay to evaluate embryotoxicity as part of the evaluation of 16 industrial chemicals. Three-day-old white Leghorn chicken eggs were injected with 1.3-10.5 µmol of TEA in acetone. Eleven days after the injection (total incubation period 14 days), eggs were opened and the embryos inspected for survival and external malformations. The ED50 for embryotoxic effects was 2.6 µmol/egg. The embryotoxic effects included early mortality and malformations (open coelom, short back or neck, edema, and lymph blebs). The incidence of malformations (3%-6%) in the TEA-treated groups was not significantly different from that of controls. GLP and study validity are not specified, however the study is clearly documented with generally accepted scientific principles considered acceptable for assessment.

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