Acetone oxime

Specific investigations: other studies

Administrative data

Endpoint:

mechanistic studies

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study meets generally acceptable scientific standards, valid for assessment. No data on GLP.

Data source

Reference

Reference Type:

publication

Title:

Amination of Tyrosine in Liver Cytosol Protein of Male F344 Rats Treated with 2-Nitropropane, 2-Nitrobutane, 3-Nitropentane, or Acetoxime

Author:

Sodum R, Fiala ES

Year:

1997

Bibliographic source:

Chem. Res. Toxicol. 1997, 10, 1420-1426.

Report date:

1997

Materials and methods

Principles of method if other than guideline:

In this paper, the authors examined liver proteins of rats treated with acetoxime for the presence of 3-aminotyrosine, the expected product of tyrosine amination. It was used ion-pair and/or cation-exchange high-performance liquid chromatography with electrochemical detection.

GLP compliance:

not specified

Type of method:

in vivo

Endpoint addressed:

carcinogenicity

Test material

Test material information

Constituent 1

Reference substance name:

Acetone oxime

EC Number:

204-820-1

EC Name:

Acetone oxime

Cas Number:

127-06-0

Molecular formula:

C3H7NO

IUPAC Name:

acetone oxime

Details on test material:

- Name of test material (as cited in study report): Acetoxime

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Taconic Labs, Germantown, NY.

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

other: Emulphor 620

Details on exposure:

Acetoxime was dissolved in an aqueous 10% solution of Emulphor 620 (GAF Corp., Wayne, NJ) for administration to rats.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

From 6 to 23 h after a single administration.

Frequency of treatment:

Only one administration.

Post exposure period:

Rats were sacrificed 6, 18, or 23 h after administration.

Doses / concentrations

Remarks:

Doses / Concentrations:
3.3 mmol/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

One male rat per time point.

Control animals:

yes, concurrent vehicle

Details on study design:

HPLC Instrumentation:
Solvent was delivered with the Shimadzu model LC-600 HPLC pumps (Shimadzu Scientific Instruments, Braintree, MA) with Shimadzu model SCL-6B solvent programmer. The eluate was analyzed using the Waters model 996 photodiode array detector and the BAS (Bioanalytical
Systems, West Lafaytte, IN) model LC-4B amperometric detector with a glassy carbon working electrode and the Ag/AgCl/3 M NaCl reference electrode. An electrode potential of 475 mV was used for analysis of protein hydrolysates and 600 mV was used for nucleoside analysis. Data were collected and processed using the Waters Millennium Chromatography Manager.

Protein Sample Preparation:
Rat livers were thawed at 4 °C and homogenized, and the cytosol supernatant was obtained by centrifugation at 107000 g. Protein concentrations in the cytosolic fractions were 25 mg/mL as determined using the Sigma protein determination kit. Protein was obtained from the cytosolic fractions by precipitation with excess acetone containing 0.2% HCl and was then dried under nitrogen. The protein was hydrolyzed under vacuum in protein hydrolysis tubes with 6 N HCl at 150 °C for 1 h; then the HCl was removed by evaporation under a stream of nitrogen. Under these hydrolysis conditions 85% of added 3-aminotyrosine could be recovered. The dry residue was redissolved in 1-2 mM sodium citrate, pH 3.5, and analyzed by ion-pair and cation-exchange HPLC with UV and EC detectors.

Examinations

Examinations:

Rats were sacrificed, and the livers were removed and frozen immediately in liquid nitrogen before storage at -80 ºC.

Results and discussion

Details on results:

It was found that the liver cytosolic proteins of these animals contained 0.53 ± 0.03 mol of 3-aminotyrosine/1E+03 mol of tyrosine.

Any other information on results incl. tables

It was found that the liver cytosolic proteins of these animals contained 0.53 ± 0.03 mol of 3-aminotyrosine/1E+03 mol of tyrosine.

Applicant's summary and conclusion

Conclusions:

It was found that the liver cytosolic proteins of these animals contained 0.53 ± 0.03 mol of 3-aminotyrosine/1E+03 mol of tyrosine.

Executive summary:

In this paper, the authors examined liver proteins of rats treated with acetoxime for the presence of 3-aminotyrosine, the expected product of tyrosine amination.

Using ion-pair and/or cation-exchange high-performance liquid chromatography with electrochemical detection, it was found that the liver cytosolic proteins of these animals contained 0.53 ± 0.03 mol of 3-aminotyrosine/1E+03 mol of tyrosine.