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Ecotoxicological information

Endocrine disrupter testing in aquatic vertebrates – in vivo

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Administrative data

Endpoint:
amphibian: other
Remarks:
Inhibition of germinal vesicle breakdown in vitro
Type of information:
experimental study
Adequacy of study:
supporting study
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:
Inhibition of germinal vesicle breakdown in Xenopus oocytes in vitro by a series of substituted glycol ethers
Author:
Fort DJ, Mathis MB, Guiney PD, Weeks JA.
Year:
2017
Bibliographic source:
J Appl Toxicol. 2017;1–10.

Materials and methods

Principles of method if other than guideline:
- Principle of test: A 24 hour in vitro Xenopus oocyte maturation (germinal vesicle breakdown [GVBD]) assay
- Parameters analysed / observed: progesterone or androstenedione binding affinities to the oocyte plasma membrane progesterone receptor (OMPR) or classical androgen receptor
- This assay is only used by a few labs and is not well established. It also doesn’t appear to have gone through a rigorous validation.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
2-butoxyethanol
EC Number:
203-905-0
EC Name:
2-butoxyethanol
Cas Number:
111-76-2
Molecular formula:
C6H14O2
IUPAC Name:
2-butoxyethanol
Specific details on test material used for the study:
Source: Sigma Aldrich
Purity: 99.4%

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
yes
Remarks:
Oocyte Ringer's buffer (pH 7.8)

Test organisms

Aquatic vertebrate type:
frog
Test organisms (species):
Xenopus laevis
Details on test organisms:
TEST ORGANISM
- Source: Dexter, MI
- Life stage: adult

ACCLIMATION
- Acclimation period: 120 days

METHOD FOR PREPARATION AND COLLECTION OF EGGS
- Animals were killed in 0.2% (w/v) 3‐aminobenzoic acid ethyl ester immediately before ovary excision

Study design

Test type:
other: in vitro

Test conditions

Nominal and measured concentrations:
0.001, 0.01, 0.05, 0.1, 0.5, 1.0, 5.0 and 10.0 mg/L. Tested in triplicate with and without the presence of 250 μM progesterone or 250 μM androstenedione:
Details on test conditions:
TEST SYSTEM
- Test vessel: six‐well glass culture plates,
- No. of organisms per vessel: 20 oocytes per replicate
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- Vehicle control performed: yes
- No. of vessels per vehicle control (replicates): 3
Reference substance (positive control):
not required
Remarks:
Mutliple substances studied. 2-methoxyethanol used as reference substance

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
other: IC25
Effect conc.:
1.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: progesterone induced GVBD inhibition
Remarks on result:
other: 0.5% of the potency of 2-methoxyethanol. 95% CI 1150 - 1650). Equiv to 1.2E-05M
Duration:
24 h
Dose descriptor:
other: IC25
Basis for effect:
other: Androstenedione‐induced GVBD inhibition potencies
Remarks on result:
not determinable
Remarks:
not reached at maximum tested concentration of 10mg/L (8.5e-05M)

Applicant's summary and conclusion

Validity criteria fulfilled:
not applicable
Executive summary:

A 24 hour in vitro Xenopus oocyte maturation (germinal vesicle breakdown [GVBD]) assay was used to screen 2-butoxyethanol for possible endocrine disruption.  ). It inhibited progesterone but not androstenedione induced GVBD although the effect was very weak (IC25 200x lower than that for 2-methoxyethanol.  The assay seems to be very sensitive (prone to false positives) compared to mammalian systems and does not provide any substantive evidence of a possible endocrine mode of action.