4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Materials and methods

Test guideline

Qualifier:

no guideline followed

Guideline:

other: A-660/3-75-009

Principles of method if other than guideline:

not applicable

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane

EC Number:

500-033-5

EC Name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane

Cas Number:

25068-38-6

Molecular formula:

(C15H16O2 . C3H5ClO)x

IUPAC Name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane

Test material form:

liquid: viscous

Details on test material:

The water soluble fraction of EPIKOTE 828 was studied. GC/Mass spectrometric analysis would suggest that the major component was the hydrolysis product of one of the epoxides.

The water soluble fraction was generated using a stainless steel column (200 x 10 mm id) packed with WHP which had been coated with EPIKOTE 828 at a loading of 5% w/w. The chromosorb was coated by evaporation from dichloromethane. A similar column was packed with uncoated chromosorb WHP and used in the preparation of control media. A new pair of EPIKOTE 828 and control columns was used to prepare the test and control media for each test.

The column was linked into a recirculating system. Dilution water was recirculated through the column from a 2 L reservoir at a rate of 2 ml/min. To monitor attainment of equilibrium concentrations of the water-soluble fraction of EPIKOTE 828 in the D. magna media preparation system, samples were taken for analysis at intervals prior to the start of the test. Since themedia preparation systems for D. magna and S capricornutum were similar it was not considered necessary to repeat this exercise for the S. capricorriutum test.


For testing, the solutions containing the water-soluble fraction of EPIKOTE 828 were diluted as appropriate to produce the required test media. The media circulated through the control columns were tested undiluted.

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Water samples were taken for analysis at intervals prior to and after the start of the test. During method development work it was shown that the water-soluble fraction of EPIKOTE 828 was composed principally of some species other than the major component of the test material as supplied. Mass spectrometry showed that, though this "water-soluble fraction" used as a standard was not pure, it contained a principal component which was probably a diol formed by hydrolysis of one of the two epoxide moieties present in the major component of EPIKOTE 828.

An analytical standard was obtained by preparative chromatography of EPIKOTE 828 to separate a quantifiable amount of the "water soluble fraction".

Three injections of EPIKOTE 828 in MTBE (3 x 100 mg EPIKOTE) were made on the following HPLC system:
Column: 25 cm x 9 mm I.D. stainless steel
Packing: Partisil 5 PAC
Mobile phase: MTBE + methanol (99.5 + 0.5 v/v)
Flow rate: 5 ml/min
Wavelength: 228 nm
(MTBE - methyl t-butyl ether)

Five fractions were collected following each injection. Fraction 5 co-chromatographed with the contents of the stock solutions and was used to produce analytical standard solutions for use on the following HPLC system:
Column: 25 cm x 4.6 mm I.D. stainless steel
Packing: Spherisorb 5 NH
Mobile phase: MTBE + methanol (92.5 + 7.5 v/v)
Flow rate: 1 ml/min
Wavelength: 228 nm
Retention time: "Water soluble fraction" ca 7.5 -8.2 min.

Test solutions

Vehicle:

no

Details on test solutions:

The "water soluble fraction" obtained from EPIKOTE 828 and used as an analytical standard in the aquatic tests was examined by mass spectrometry. Though it was not possible to characterize this material completely, the results suggested that a major component of the "water soluble fraction" was the diol presumably formed by hydrolysis of one of the two epoxide function of DGEBPA, the major component of EPIKOTE 828

Test organisms

Test organisms (species):

Scenedesmus capricornutum

Details on test organisms:

S. capricornutum were taken from the axenic laboratory culture. The culture is derived from a strain (ATCC 22662) obtained from the American Type Culture Collection, Maryland, U.S.A..

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Post exposure observation period:

Study was stopped after 72 hours and no further observations made

Test conditions

Hardness:

no data

Test temperature:

24-25C

pH:

pH for the control group (which had not been passed through a chromosorb column) ranged from 7.1 at the start of the study to 8.6 at the end of the study. pH for the top concentrations ranged from 7.4 at the start of the study to 7.5 at the end of the study.

Dissolved oxygen:

no data

Salinity:

no data

Nominal and measured concentrations:

Nominal: 0 (control), 0 (chromosorb control), 2.2, 4.6, 10, 22, 46, 100
Measured: <0.01, <0.01, 0.18, 0.48, 1.1, 2.4, 4.2, 11

Details on test conditions:

A 3 day (72 h) growth test was carried out. A saturated stock solution of the water-soluble fraction of EPIKOTE 828 in algal growth medium was produced. Twenty-seven, 250 ml Erlenmeyer flasks were prepared. To five triplicate sets of these were added quantities of the stock solution and algal growth medium sufficient to produce, 46, 22, 10, 4.6 and 2.2% dilutions of the stock solution when made up to a final volume of 50 ml. A sixth set of three flasks was part filled with 50 ml of the undiluted stock solution. Two sets of control flasks were also prepared. The first set, consisting of six flasks, were part filled with 50 ml of algal growth medium. The second set, consisting of the remaining three flasks, were part filled with 50 ml of algal growth medium that had been circulated through a column packed with Chromosorb WHP that had not been coated with EPIKOTE 828. The latter also served as controls and are subsequently referred to as chromosorb controls.

Each flask was inoculated with sufficient S. capricornutum cells to give an initial nominal cqncentration of 10 000 cells ml-. The flasks: were then closed with a loose fitting aluminium cap and incubated in a cooled orbital incubator (100 cycles/min) under constant illumination (~3000 lux) at a nominal temperature of 22-26°C for 3 days (72 h). At 0 h and then at 24 h intervals after the start of incubation cell counts were made using a Coulter Multisizer. The pH in the control and highest concentration test solutions were determined at the start and conclusion of the test. The incubation temperature was monitored throughout the test.

Samples of the freshly prepared and 72 h old test and control solutions were analysed to determine the concentration of the water-soluble fraction of EPIKOTE 828.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

ca. 9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: water soluble fraction

Basis for effect:

biomass

Remarks on result:

other: 5.6 - 42 mg/L

Details on results:

Inhibition of the growth of S. capricornutum by more than 50% was only detected at the highest test concentration when the data were analysed in terms of the area under the growth curves (A) and then only after 48 and 72 hours of exposure.

The highest concentration in which growth of S. capricornutum was not inhibited over the 72 hour test period was 2.4 mg/L when growth was expressed in terms of A and 4.2 mg/L when growth was expressed in terms of the average specific growth rate (u).

Results with reference substance (positive control):

no data

Reported statistics and error estimates:

see above.

Any other information on results incl. tables

The 48 and 72 hour EC50 values were 9.1 and 9.4 mg/L, respectively.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48 and 72 hour EC50(biomass) values were 9.1 and 9.4 mg/L, respectively.

Executive summary:

The toxicity of EPIKOTE 828 to S. capricornutum was determined in a 72 h growth test. Based upon analyses of the test results in terms of the area under the culture growth curves the 48 and 72 EC50 values were calculated to be 9.1 and 9.4 mg/L, respectively, when expressed in terms of the concentration of the water-soluble fraction of EPIKOTE 828.