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Toxicological information

Basic toxicokinetics

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Administrative data

Endpoint:
basic toxicokinetics in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was performed in 1960, so the report is not very detailed. However, the report contains sufficient information to consider the study and the results valid.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1960

Materials and methods

Objective of study:
other: Determination of degree of hydrolysis of erucamide to erucic acid and ammonia
Principles of method if other than guideline:
Erucamide was incubated with rat liver homogenate to determine the degree to which erucamide is hydrolyzed to erucic acid and ammonia. The production of ammonia was measured using the aeration technique of Van Slyke and Cullen (J. Biol. Chem. 24: 117, 1916).
GLP compliance:
no
Remarks:
study was conducted prior to GLP regulations

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Test material described only as "a purified grade of erucamide". Associated documents suggest the composition was 98.5% erucamide, 0.8% free fatty acid (as oleic), 0.5% moisture and 0.2% nitrile (as oleyl).
Radiolabelling:
no

Test animals

Species:
rat
Strain:
not specified
Sex:
not specified
Details on test animals and environmental conditions:
None

Administration / exposure

Route of administration:
other: in vitro incubation
Vehicle:
other: test material dissolved in chloroform which was then removed by evaporation
Doses / concentrations
Remarks:
Doses / Concentrations:
3.6 mg erucamide per vessel
Details on study design:
The fatty acid amidases which occur in rat liver were chosen as typical of mammalian amidases and used in the hydrolysis investigation. A known quantity of erucamide was dissolved in chloroform so that exactly 3.6 mg could be introduced into a Warburg vessel in 2 ml of solution. The solvent was evaporated with a stream of nitrogen, leaving a film of the amide on the inside surface of the flask.

Liver from a freshly killed, exsanguinated rat was homogenized and 0.5 g of the liver homogenate added to the pre-weighted Warburg flask. Two ml of Ringer-phosphate medium was then added and 0.2ml of 2.5 N HCl pipetted into the side arm and well of the flask. Incubation at 37 °C and agitation was carried out for four hours. At the end of this time, the HCl was tipped into the mixture which was then transferred to a test tube containing 1 ml of 15 per cent trichloroacetic acid. After centrifugation, the supernatant liquid was drawn off and the precipitate washed two or three times with 3 per cent trichloroacetic acid which was combined with the original solution.

Toal ammonia was determined using the aeartion technique of Van Slyke and Cullen. Inasmuch as ammonia is formed naturally by liver homogenate, blank experiments were conducted on all constituents except the erucamide.

Results and discussion

Metabolite characterisation studies

Metabolites identified:
no

Any other information on results incl. tables

Volume of 0.02 N HCl consumed by sample: 0.55 ml

Volume of 0.02 N HCl consumed by blank: 0.35 ml

Ammonia liberated (micromoles): 4.0

Initial amount of sample (micromoles): 10.65

Degree of hydrolysis (per cent): 37.6

Applicant's summary and conclusion

Conclusions:
Erucamide was found to be efficiently hydrolyzed by rat liver homogenate, with 37.6 per cent hydrolyzed in four hours.
Executive summary:

Erucamide was incubated with rat liver homogenate to determine the degree to which erucamide is hydrolyzed to erucic acid and ammonia. Erucamide was found to be efficiently hydrolyzed by rat liver homogenate, with 37.6 per cent hydrolyzed in four hours.