4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with [(dimethylamino)methyl]phenol and piperazine

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

This study was conducted between 08 December 2016 and 10 February 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese notification, Yakushokuhatsu 0331 No.7, Heisei 23.03.29 Seikyoku No.5, Kanpokihatsu No.110331009, March 31, 2011

Version / remarks:

the latest revision, December 21, 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name of the newchemical substance(by IUPAC nomenclature) 4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with [(dimethylamino)methyl]phenol and piperazine
Alternate name THEMIS
CAS RN 159034-96-5
Molecular weight Mn:3,000，Mw:11,000
Purity (%) of the new chemical substance subjected to the study 100.0
Lot number of the new chemical substance subjected to the study 160715
Melting point 120 to 140°C
Appearance at room temperature Powder, pale pink
Stability Swell on exposure to water. Soften at greater than or equal to 100°C

Analytical monitoring:

yes

Details on sampling:

Samples of the test solutions at the beginning and the end of the renewal period, i.e. when it is freshly prepared and immediately prior to renewal or at the end of exposure are referred to as one set. The two sets of test solutions, one at the beginning of exposure (0 hour) and before renewal at 24 hours and the other after renewal at 72 hours and at the end of exposure (96 hours), were analyzed.

Vehicle:

no

Details on test solutions:

Preparation of Test Solutions
1. Loading of the test substance
0.520 g Test substance was collected to a 5-L media bottle and the dilution water was added to final volume of 5.2 L. The preparation frequency were 48 hours before the start of exposure and 48 hours before renewal of test solutions.
2. Mixing method
Stirred with a magnetic stirrer for 48 hours (room temperature, approx. 24°C).
3. Collection of WAF
Filtrated the stirred solution with filter (Qualitative fùter paper No. 2) and then transfered the filtrates (5 L) to the test vessel, and used for the test.
(Test solutions were aerated before use in order that dissolved oxygen concentration would reach 7.5 mg/L or more.)

Test organisms (species):

Oryzias latipes

Details on test organisms:

Test Species
1) Common name: Medaka
2) Scientific name: Oryzias latipes
3) Source: Breeding in the testing facility
4) Date of starting the collection of eggs: June 20, 2016
5) Lot No.: 16-M-0620B
6) Size: Total length: Recommended: 2.3 ± 1.2 cm
Measurements: 2.35 cm (2.02 – 2.76 cm), n=10
Body weight: Measurements: 0.116 g (0.085 – 0.232 g), n=10
(These values were measured and weighed for the fish of control at the end of exposure.)
7) Age: Less than a half year after hatching
8) Medication before acclimation: Applied (New Green F, San-ei Pharmaceutical Co., Ltd.)
9) Acclimation period: November 10, 2016 -December 12, 2016
10) Acclimation conditions:
Water: Dechlorinated water
Procedure: Flow-through (Biomass loading: 1 g fish/Llday or less)
Temperature: 24 ± 1 °C
Dissolved oxygen concentration (D.O): 80% or more of the saturation
pH: 6.0 - 8.5
Light: Fluorescent light, 16-hour light (1000 lux or less) / 8-hour dark
Feed: TetraMin, TetraWerke
Feeding amount: 2% ofthe body weight/day (approx.) (until 24 hours before exposure)
Medication during acclimation: Not applied
Mortality during 7 days before the test: Less than 5%

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

23.7 - 24.1 °C

pH:

6.9 -7.6

Dissolved oxygen:

6.2 - 8.2 mg/L

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: 6.43 mg/L

Details on test conditions:

Test Conditions
1) Duration of exposure: 96 hours
2) Exposure procedure: Semi-static (Batchwise renewal of the test solution after every 24-hour exposure. )
3) Volume of test solution: 5 L/vessel (The measurement samples of water quality and analysis were collected from each vessel; kept biomass loading: 1 g fish/L or less)
4) Number of vessels: 1 vessel/test group
5) Biomass loading: 10 fish/ test group
6) Temperature: 24±1°C
7) D.O.: 60% or more of the saturation (not aerated)
8) pH: Not adjusted
9) Light: Fluorescent light, 16-hour light (1000 lux or less) / 8-hour dark
10) Feeding: None

Results of Range-finding Test
A range-finding test was conducted at loading rates below the upper limit concentration (100 mg/L) given in the Test Guideline. A water-accommodated fraction was used as a test solution because the test substance was multi-component substance and poorly soluble in water. No mortality and none toxicological symptoms of living fish was observed at 96 hours for 100 mg/L.
Test condition: 5 fish/2.5L/test group

Determination of Test Concentrations
Based on the results of the range-finding test, the concentrations for the definitive test were determined to be 100 mg/L (loading rate).

Test Procedure
The acclimated fish were loaded randomly to the test solutions, according to a random table, at the start of exposure. To maintain the concentration on the test substance, test solutions were renewed every 24 hours. Renewals of the test solutions were performed by transferring the fish to freshly prepared test solutions. Observations and measurements were performed as described below.

1) Measurements of quality and observation of appearance of the test solutions
Measurements of the temperature, dissolved oxygen (D.O.), and pH, and observation of the appearance of the test solutions were performed once a day. When renewing the test solutions, measurements and observations were carried out on both the freshly prepared test solutions and the test solutions immediately prior to renewal.

2) Observation of test fish
Test fish were observed at 3, 24, 48, 72 and 96 hours after the start of exposure.

Classification and definition of toxicological symptoms
Death: No visible movement (e.g., gill cover movement) and touching of the caudal peduncle produces no reaction.
Critical:
Impossible to swim: Although the gill cover moves, no other movement (e.g., swimming) or extreme abnormalities are observable.
Abnormal:
Abnormal swimming: Swimming behavior is not normal (e.g., inactive, hyperactive, surfacing, loss of equilibrium, erratic, twitching).
Abnormal appearance: Appearance is not normal (e.g., light discoloration, dark discoloration, hemorrhage, erosion, mucous coagulation, protrusion of scale, loss of scale, loss of fin, curved spine, exophthalmoses).
Abnormal respiration: Gill cover movements are not normal (e.g., rapid, slow, irregular).
Normal: No abnormality in swimming behavior, appearance, and so on.

Reference substance (positive control):

yes

Remarks:

Copper sulphate

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 6.43 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Details on results:

Quality and Appearance of Test Solutions
Temperatures ranged from 23.7 to 24.1°C, the D.O. ranged from 6.2 to 8.2 mg/L, and pH ranged from 6.9 to 7.6. In both the control and Conc. 1, the test solutions appeared colorless, and suspended substances, floating substances and precipitated substances were not observed in the test solutions during exposure.

Observation of Test Species
In both the control and Conc. 1, no dead fish and no abnormal symptoms in living fish were observed.

Median Lethal Concentration (LC50) and Median Lethal Loading Late (LL50)
The LC50 and LL50 values of each period are shown in Table 7, and that of 96-hour exposure is given below.
96-hour LC50: >6.43 mg/L (95% confidence limits: not available)
96-hour LL50: >100 mg/L (95% confidence limits: not available)

Validity of the Test
This test is valid because the results satisfied the requirements of the test.

Results with reference substance (positive control):

The sensitivity of test fish has been confirmed in each lot by the acute toxicity test of reference substance (CuSO4(II)∙5H2O, guaranteed reagent).
The 96-hour median lethal concentration (96-hour LC50, in anhydrous) is shown below.
96-hour LC50 of the lot concerned (date of performing the acute toxicity test: October 7, 2016)
0.26 mg/L (95% confidence limits: 0.10 – 0.40 mg/L)

The average LC50 value in this laboratory (n = 80, since 1995)
Average ± standard deviation: 0.73 ± 0.29 mg/L
Minimum value – maximum value: 0.28 – 1.5 mg/L
The LC50 of reference substance fell within the standard range defined by the testing facility (average of LC50 of lots in the past ± 5 times of the standard deviation).

Validity criteria fulfilled:

yes

Conclusions:

The result given below was calculated from the time-weighted mean of measured concentrations and the loading rate. 
           96-hour LC50: >6.43 mg/L (95% confidence limits: not available)
           96-hour LL50: >100 mg/L (95% confidence limits: not available)

Executive summary:

A semi-static test was conducted in accordance with OECD Guideline for Testing of Chemicals No.203 “Fish, Acute Toxicity Test” (1992).

The test species Medaka (Oryzias latipes) was exposed to the test substance at a loading rate of 100 mg/L for 96 hours.

 

The result given below was calculated from the time-weighted mean of measured concentrations and the loading rate.

 

           96-hour LC50: >6.43 mg/L (95% confidence limits: not available)

           96-hour LL50: >100 mg/L (95% confidence limits: not available)

Description of key information

A semi-static test was conducted in accordance with OECD Guideline for Testing of Chemicals No.203 “Fish, Acute Toxicity Test” (1992).

 

The result given below was calculated from the time-weighted mean of measured concentrations and the loading rate.

 

           96-hour LC50: >6.43 mg/L (95% confidence limits: not available)

           96-hour LL50: >100 mg/L (95% confidence limits: not available)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The registered substance is insoluble with water solubility of 1mg/L at 20℃. The test results showed 96h LL50>100mg/L which indicating no hazard identified at highest loading rate. Therefore LC50=100mg/L is used for CSA for worst cases.