Reaction products of 4,4'-methylenebis(cyclohexylamine) and 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Samples for possible analysis were taken from all test concentrations and the control according to the schedule below. In addition samples were taken from the acetone stocks.
Frequency at t=0 h and t=48 h from the test solutions
at t=0h from stock solutions
Volume 50 mL from the approximate centre of the test vessels (test solutions)
3 mL from stock solutions
Storage Samples were stored in a freezer until analysis.
At the end of the exposure period, the replicates were pooled at each concentration before sampling.
Additionally, reserve samples of 50 mL (test solutions) and 3 mL (stock solutions) were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.

Vehicle:

no

Remarks:

See below the details on test solution

Details on test solutions:

Preparation of test solutions
The batch of PACM BADGE adduct tested was a clear light yellow solid UVCB substance. No correction was made for the purity/composition of the test item.
In order to obtain a homogeneous sample, the test item was heated for approximately 3.5-5.5 hours at 90ºC prior to the preparation of test solutions. After warming, the appearance of the test item changed from a clear light yellow solid to a clear viscous liquid.
First combined limit/range-finding test and full tests: Preparation of test solutions started with individually prepared loading rates ranging from 1.0 to 100 mg/L. The medium used was heated to 70°C before the test item was added. A 3-day period of magnetic stirring was applied to accelerate dissolution of the test item in medium. Thereafter, the aqueous Water Accommodated Fractions (WAFs) were filtered and used as test concentrations. The final test solutions were clear and colorless.
Second combined limit/range-finding test: stock solutions were prepared in acetone (100%, VWR France) at loading rates of 10, 100 and 1000 mg/L. An overnight period of magnetic stirring followed by a 13-minute period of ultrasonic waves was applied. The stock solutions were clear and colorless. Subsequently 100 µL of each stock was spiked at 1 liter of continuously stirred medium. The obtained solutions were clear and colorless.
M7, as prescribed by Dr. Elendt-Schneider
(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).

Test organisms (species):

Daphnia magna

Details on test organisms:

Species Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
Source In-house laboratory culture with a known history.
Reason for selection This system has been selected as an internationally accepted invertebrate species.
Validity of batch Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Characteristics For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Macro salts: CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L

Trace elements: B 0.125 mg/L
Fe 0.05 mg/L
Mn 0.025 mg/L
Li, Rb and Sr 0.0125 mg/L
Mo 0.0063 mg/L
Br 0.0025 mg/L
Cu 0.0016 mg/L
Zn 0.0063 mg/L
Co and I 0.0025 mg/L
Se 0.0010 mg/L
V 0.0003 mg/L
Na2EDTA.2H2O 2.5 mg/L
Macro nutrients: Na2SiO3.9H2O 10.0 mg/L
NaNO3 0.27 mg/L
KH2PO4 0.14 mg/L
K2HPO4 0.18 mg/L
Vitamins: Thiamine 75.0 µg/L
B12 1.0 µg/L
Biotin 0.75 µg/L
The hardness: 180 mg/L expressed as CaCO3 and the pH: 7.7 ± 0.3.

Test temperature:

18-22 C

pH:

7.7-7.8

Dissolved oxygen:

8.8 mg/l

Nominal and measured concentrations:

Analysis of the samples taken at the start of the test showed measured concentrations of 0.43 and 0.41 mg/L in the solvent control and the limit concentration of 0.10 mg/L. The concentration measured in the limit group was four time higher than the nominal concentration and below the limit of quantification of the analytical method, thus the results were considered not reliable.
The concentration measured in the stock solution prepared at 1000 mg/L was at the level of nominal confirming proper preparation of test solutions. Based on this result, the effect parameters were expressed in terms of nominal concentrations.

Reference substance (positive control):

yes

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The responses recorded in this test allowed for reliable determination of an EC50. No immobilization or other effects were observed in any of the groups tested during the exposure period. This shows that the test item was not toxic to daphnid at its solubility limit in medium.

Validity criteria fulfilled:

yes

Conclusions:

Daphnia acute immobilization test was performed with PACM BADGE adduct. In conclusion, the 48h-EC50 was beyond the range tested, i.e. exceeded a nominal concentration of 0.10 mg/L being considered maximum soluble in test medium. Result shows that the test item was not toxic to daphnid at its solubility limit in medium.

Executive summary:

Daphnia acute immobilization test was performed with PACM BADGE adduct. In conclusion, the 48h-EC50 was beyond the range tested, i.e. exceeded a nominal concentration of 0.10 mg/L being considered maximum soluble in test medium. Result shows that the test item was not toxic to daphnid at its solubility limit in medium.

Description of key information

A Daphnia acute immobilization test was performed with PACM BADGE adduct. In conclusion, the 48h-EC50 was beyond the range tested, i.e. exceeded a nominal concentration of 0.10 mg/L being considered maximum soluble in test medium. Result shows that the test item was not toxic to daphnid at its solubility limit in medium.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

0.1 mg/L