1,2-Cyclohexanedicarboxylic Acid, 1-(phenylmethyl) ester, ester with 2,2,4-trimethyl, 1,3-petanediol mono(2-methyl propanoate)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17/11/2015-23/02/2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Principles of method if other than guideline:

The latest water characterisations in June 2015 were not performed according to GLP. The test article characterisation was not performed according to GLP. THese were the only exceptions to conforming to GLP principles and did nmot adversely affect study integrity or interpretation of the study results.

GLP compliance:

yes

Specific details on test material used for the study:

Santicizer P1700 Cas No 1661012-65-2 Lot No SC 1002-2 Purity 96.86%

Analytical monitoring:

yes

Details on sampling:

Test solution samples of Santicizer P1700 were collected at (initiation) 0, 48 and 96 (termination) of the definitive test.

At each sampling point, 10mL was collected using a serological pipette, from control, vehicle control and tst substance treatment and placed in separate 15mL cul;ture tubes. SAmples were diluted with 2mL cyclohexane and votexed for 1 min to mix. After the water and cyclohexane phases separated the top cyclohexane layer was transferred back into a separate container. A second 2mL volume of cyclohexane was added, shaken, vortexed and allowed to separate. THe cyclohexane layer was again removed and combined with the initial extract sample. If needed the samples were further diluted with cyclohexane to bring them within the analytrical standards range (2-100ug/L. Two QC fortification spikes that bracket expected high and low treatmenmt concentrations of the test substance P1700 were prepared amd analysed in a similar manner.

Sample analysis was by GC/MS

Vehicle:

yes

Remarks:

DMF

Details on test solutions:

Based on the results of the range finding and visual solubility tests, a static definitive test was performed from 23 to 27 November 2015 at nominal concentrations 0, ad 1.8mg/L Santicizer P1700. A primary standard was prepared at initiation at a concentration of 18mg/mL Santicizer P1700 by dissolving 0.0929g Santicizer P1700/mL (0.0900g corrected for 96.86% purity) in 5mL dimethylformamide (DMF). The 1.8mg Santicizer P1700/L test solution was prepared by adding 1.8mL of the primary standard solution (in D~MF) to a volume of 18L with dilution water. The vehicle control was 1.8mL DMF in 18L dilution water. The control was dilution water only.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Rainbow trout (Oncorhynchus mykiss) were obtained from Trout Lodge, Sumner, Washington. Fish were fed salmon starter daily. Fish were maintained for at least 7 days in dilutiuon water at a temperature within ±2°C of the test temperature. No diseased were observed or treated during the 14 day period prior to use in the test. Food was witheld during acclimation fo approximately 48 hours prior to initiation of the test, and mortality did not exceed 5% during the 48hours immediately before definitive test initiation. A representative sample of ten acclimated fish were measured at test initiation and ranged from 38-45mm total length (mean 42 SD ±2.1 mm). Blotted wet weight range 0.4029 to 0.6831 g (mean 0.5639 SD ±0.0831 g). Based on this representative sample the instantaneous biomass loading rate was 0.22g fish tissue per litre of test solution, which is below the protocol maximum of 0.8g fish tissue/ L.

For the definitive test, 7 fish were impartially assigned to treatments by adding one fish per chamber, first control then low to high substance treatments and repeating steps as neceassary until 7 fish were present in each test chamber. The treatments were not replicated. Observations for mortality and sublethal responses were made at approx 24, 48, 72 and 96 hours. No food was provided during the 96hr exposure period.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

0 (96 hr while exposed).

Hardness:

160mg CaCO3/L

Test temperature:

12.8-13.3 degrees C

pH:

8.1-8.5

Dissolved oxygen:

8.2-9.8 mg/L (81-97% saturation)

Salinity:

150mg/L CaCO3 (total alkalinity)

Conductivity:

341uS/cm

Nominal and measured concentrations:

Nominal 1.8mg/L Santicizer P1700
Measured 1.59mg/L

Details on test conditions:

Temperature, sissolved O2 and pH were measured daily using a WTW Multi-Meter 3430 or a WTW Mult 3500i multimeter. In addition a continuous record of the temperature from the water bath was also maintained. Total hardness and alkalinity of the dilution water were measured using titrimetric methods adapted from standard methods. Conductivity of the dilution water was measured with a WTW Cond 3300i meter.

Reference substance (positive control):

yes

Remarks:

The test substance was used for all reference spiked solutions

Key result

Duration:

96 h

Dose descriptor:

LC50

Remarks:

highest conc tested

Effect conc.:

> 1.59 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality (fish)

Remarks on result:

other: Also applies to 24hr and 48hr LC50

Key result

Duration:

96 h

Dose descriptor:

NOEC

Remarks:

highest conc tested

Effect conc.:

1.59 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality (fish)

Details on results:

There was no mortality or sublethal effects at any concentration tested (up to 1.59mg/L).

Results with reference substance (positive control):

No effects up to 3.0mg/L (in the range finding test) but solubility restrictions in water. The test substance was also used as the "reference substance" for standards and spiked recovery solutions.

Reported statistics and error estimates:

Due to no lethal or sublethal effects, statistical analyses were not performed.

Conclusions:

There was no mortality among control animals during the study. Therefore control animals satisfied test acceptability criteria for survival (i.e. >= 90% or not more than one mortality) as stated in the study protocol and OECD 203 testing guidelines.

Based on mean measured Santicizer P1700 concentrations the estimated 24, 48 72 and 96 hr LC50 values were >1.59mg/L Santicizer P1700, the highest concentration tested. The 96hr NOEC was 1.59mg/ L Santicizer P1700, based on mean measured concentrations and a lack of any mortality or sublethal effects.

Executive summary:

There was no mortality among control animals during the study. Therefore control animals satisfied test acceptability criteria for survival (i.e. >= 90% or not more than one mortality) as stated in the study protocol and OECD 203 testing guidelines.  

Based on mean measured Santicizer P1700 concentrations the estimated 24, 48 72 and 96 hr LC50 values were >1.59mg/L Santicizer P1700, the highest concentration tested. The 96hr NOEC was 1.59mg/ L Santicizer P1700, based on mean measured concentrations and a lack of any mortality or sublethal effects.  

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 15 to March 19 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP compliant (no certificate), near guideline study, available as unpublished report, minor limitations in design and reporting but otherwise adequate for assessment.

Justification for type of information:

The justification for read across is provided as an attachment in IUCLID Section 13.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The deviations did not impact the result of the study

Principles of method if other than guideline:

Procedures as described by US EPA and American Public Health Association were followed (1979). Other procedure: ABC Protocol #7601

GLP compliance:

yes

Specific details on test material used for the study:

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

Analytical monitoring:

no

Details on sampling:

No data

Vehicle:

no

Details on test solutions:

ABC Lab well water. Test concentrations were prepared by weighing the indicated amounts of S278 and transferring directly to the test chambers. The sample container was rinsed with 10ml acetone followed by two rinsings in the test dilution water. The control chamber received a 10ml aliquot of acetone. An oily film was noted on the surface of the test chambers following toxicant additions.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

Obtained from Spring Creek Trout Hatchery, Lewistown, Montana. Shipped by air and received February 21 in excellent condition. Mean weight: 0.63g, mean standard length: 32.6mm. (Measurements made at termination of test.) All test fish were held in culture tanks on a 16-hour daylight photoperiod and observed for at least 14 days prior to testing. Daily records were kept, including any prophylactic and therapeutic disease treatments. Fish were fed a standard commercial fish food daily. The test fish were acclimated to the dilution water and test temperature (12°C) and held without food 48 hours prior to testing.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

not reported

Hardness:

240mg/l CaCO₃

Test temperature:

12°C

pH:

7.8

Dissolved oxygen:

8.9mg/l

Salinity:

Not applicable

Nominal and measured concentrations:

two replicates of 1000 mg/l plus a control

Details on test conditions:

Preliminary test concentrations set at 1, 10, 100 and 320 mg/l and resulted in no mortality. Definitive test conducted at 1000mg/l at customer's request. Letter authorizing deviation was included in appendix.

Reference substance (positive control):

yes

Remarks:

Antimycin A

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality observed at test concentration (1000mg/l), or at preliminary range-finding concentrations (1, 10, 100 and 320 mg/l). Observation that fish were 'stressed' after 48 hours at 320 concentration (in range-finding study).

Results with reference substance (positive control):

96-hour LC50: 0.000032mg/l (C.I. 0.000027-0.000038 mg/l). The observed 96-hour LC50 and 95% confidence limits were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition.

Reported statistics and error estimates:

Probit analysis. 95% Confidence Intervals.

Sublethal observations / clinical signs:

LC50 in mg/l (ppm)
Compound	24 hrs	48 hrs	96 hrs
S278	>1000	>1000	>1000
Antimycin A	0.00021	0.000034	0.000032
95% C.I.(Antimycin A)	0.00010 -0.00043	0.000027 -0.000042	0.000027 -0.000038

Validity criteria fulfilled:

yes

Conclusions:

The LC50 was >1000 mg/l. The authors note that the concentration tested (1000 mg/l) was above the solubility of the material in water. It is concluded that S278 is non-toxic to fish at concentrations well above environmental levels.

Executive summary:

This test was conducted prior to the implementation of OECD guidelines, but conformed to accepted procedures of the time (USEPA and APHA), and is equivalent to OECD 203. It is therefore considered suitable for assessment as the key study for this endpoint.

Description of key information

In a key OECD Guideline 203 study, based on mean measured Santicizer P1700 concentrations, the estimated 24, 48 72 and 96 hr LC₅₀ values were >1.59 mg/L Santicizer P1700, the highest concentration tested. The 96hr NOEC was 1.59 mg/ L Santicizer P1700, based on mean measured concentrations and a lack of any mortality or sublethal effects (ABC Laboratories Ltd, 2016c).

Read across data from the source dossier that tested structural analogue Santicizer S278 ( benzyl 3-(isobutyryloxy)-1-isopropyl-2,2-dimethylpropyl phthalate) based on analogue is also available. A discussion and report on the read across strategy is provided as an attachment in Section 13 of the dossier.

The 96-hour LC₅₀ for Rainbow trout was >1000 mg/L, at 12°C and for Fathead minnows was >1000 mg/L, at 22°C for Santicizer 278 and is therefore considered to be non-toxic to fish at its limit of solubility.

Additional information

Two proprietary short term toxicity tests to fish were conducted using Santicizer 278, and these are used as Key and Supporting Studies for this endpoint. The 96-hour LC50 for Rainbow trout (Analytical Bio Chemistry Laboratories, 1979a) was >1000 mg/l (at 12°C), and for Fathead minnows (Analytical Bio Chemistry Laboratories, 1979b) it was also >1000 mg/l (at 22°C). In both tests the concentration used was 1000 mg/l. This concentration was determine by range finding tests to be appropriate as moratlities did not occur below this concentration.

 

This exceeds the solubility of the test compound (0.0015 mg/l at 25°C) but since no mortalities occurred even at this concentration, Santicizer 278 is considered non-toxic to fish at its limit of solubility. 

 

In addition, QSAR (ECOSAR, USEPA 2008) can predict LC50 for fish up to a log Kow of 5.0, above which it predicts no effects at saturation. Santicizer 278 has a log Kow of 7.0 so no effects would be expected according to the model.