3,6,9-trioxaundecamethylene bis(2-ethylhexanoate)

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:

March 2, 1993 to March 6, 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

The test was conducted by recognized guidelines acceptable at the time of the study.

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1400 (Fish Acute Toxicity Test)

Version / remarks:

U. S.. EPA-TSCA, 40 CFR, Part 797

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

The sample upon receipt was observed to be an amber liquid and was stored at room temperature. Sample purity was not specified. The test material sample (batch #010241) used for the third definitive attempt and the definitive was received from The C P . Hall Company on February 16, 1993, in good condition and was issued ABC reference #TS-6321.

Analytical monitoring:

no

Vehicle:

yes

Remarks:

acetone

Details on test solutions:

The definitive test concentrations were obtained by transferring appropriate weights of test compound to the test chambers. Before addition to the test chambers, 1.5 mL of acetone was added to each sample to increase dispersion of the compound in the dilution water. The solutions were then stirred overnight in an attempt to reach water solubility. The solutions were stopped from stirring to allow the undissolved compound to collect on the surface. The solutions were then siphoned into the test vessels, taking care to leave ah undissolved compound in the vessel in which the solutions were stirred. All test concentrations were based on the total compound, i.e.not corrected for sample purity. The vehicle blank chamber received a 1.5 ml aliquot of acetone, which was equivalent to the highest amount used in any test solution.Records of all standard weights and dilution values were recorded.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The 70 rainbow trout (lot #6092) used in the test were obtained from Mt. Lassen Trout Farm in Red Bluff, California. The fish were received as green eggs and sperm and were fertilized at ABC Laboratories on December 1, 1992. After hatching, the fish were reared and maintained in well water and were fed newly hatched brine shrimp (Artemia sp.) or a commercially available fish food daily. The fish food was received from Zeigler Bros., Inc., Gardners, Pennsylvania, and the brine shrimp was received from San Francisco Bay Company, Newark, California.

The fish were placed in appropriate culture tanks to acclimate to the test temperature range and test dilution water approximately 12 weeks before the test. All test fish were held on a 16-hour daylight photoperiod with a 30-minute transition period and were observed for at least fourteen days prior to testing. Fish culture techniques were basically those described by Brauhn and Schoettger (5). A daily record of fish observations during the holding period, along with any prophylactic or therapeutic disease treatments, was maintained.

A sublot of fish was acclimated to the test dilution water and test temperature approximately 48 hours prior to the test. The fish were not fed during the acclimation and test periods. Weight and length measurements were made on the control group of fish at the termination of the test. The rainbow trout (Oncorhynchus mykiss) used as the control group for this experiment had a mean wet weight of 0.80 (+ 0.29) g and a mean standard length of 40 (± 5) mm. This gave a test chamber loading biomass of 0.27 g/L for the definitive study. The control group was representative of all the fish used in the definitive study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

148 mg/L, (hardness)

Test temperature:

See Table below

pH:

See Table below

Dissolved oxygen:

See Table below

Salinity:

Not reported

Conductivity:

340 uMhos/cm (conductivity)

Nominal and measured concentrations:

Nominal Test Concentrations: Control, Vehicle Blank, the water soluble fractions (WSF) of 100, 180, 320, 560, and 1000 mg/L

Details on test conditions:

The static fish bioassay was conducted in five gallon glass vessels containing 15 L of hard blended water. This dilution water was prepared by blending naturally hard well water with well water that had been demineralized by reverse osmosis. This blended water was prepared to yield a total hardness of 130-160 mg/L as CaC03 . The 0-hour measured control water parameters of this dilution water were dissolved oxygen 9.6 mg/L and pH 8.2. The test vessels were kept in a water bath at 15 (+ 2°C).

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

There was insufficient mortality (<37% to >63%) for the calculation of an LC5 0 using the LC5 0 computer program, therefore, this value is the study director's estimate based on the mortality data.

Details on results:

The 96-hour static toxicity to rainbow trout (Oncorhynchus mykiss) exposed to the test material were determined. The 24-, 48-, 72-, and 96-hour LC50 values for TegMeR® 804 were > the WSF of 1000 mg/L, > the WSF of 560 mg/L, > the WSF of 320 mg/L, and > the WSF of 100 mg/La , respectively. All results were based on the WSFs of the nominal concentrations of 100, 180, 320, 560, and 1000 mg/L. All test solutions were clear at 0 and 24 hours. At 48 and 72 hours however, all solutions were clear except the 100 mg/L B replicate and the A and B replicates of the 320 mg/L test solutions, where a slight amount of compound was on the surface. The 560 mg/L B and the 1000 mg/L A replicates were noted as having oily droplets present on the surface. At 96 hours, there were oily droplets noted on the surface of the 100 mg/L B, 560 mg/L A, and 1000 mg/L B test solutions. All other solutions were clear 96 hours.

The 96-hour NOEC was < the WSF of 100 mg/L, the lowest concentration tested, based on mortality and/or observed abnormal (sublethal) effects at this concentration. Abnormal effects of mortality, surfacing, loss of equilibrium, dark discoloration, fish on the bottom of the test chamber, labored respiration, erratic swimming, and/or quiescence were observed during the 96-hour exposure period in the 100, 180, 320, 560, and 1000 mg/L test concentrations. The 100, 180, 320, 560, and 1000 mg/L WSF test concentrations elicited 20, 0, 30, 50, and 50% cumulative mortality, respectively, during the 96-hour exposure period. An examination of the fish culture and acclimation records for this test indicated that the fish were in good condition for testing.

The dissolved oxygen concentrations ranged from 2.1 to 9.6 mg/L during the test. These values represented 22 and 97% saturation at 15 and 14°C, respectively. The dissolved oxygen concentrations in the control replicates ranged from 6.7 to 9.6 during the test. These values ranged from 69 to 97% saturation at 15 and 14°C, respectively, and were considered adequate for testing (3), therefore, the test chambers were not aerated during testing. Low dissolved oxygen values seen in the test chambers containing TegMeR® 804 appear to be caused by the test material, since low dissolved oxygen values were not noted in the control or vehicle blank. The pH values ranged from 7.4 to 8.2. Temperature values measured continuously in the water bath during testing ranged from approximately 13.7 to 14.7 °C.

Sublethal observations / clinical signs:

Static Acute Toxicity of the test material to Rainbow Trout (Oncorhynchus mykiss)

LC50 in mg/L

Compound	24 Hour	48 Hour	72 Hour	96 Hour
Test Material	> the WSF of 1000 (b)	> the WSF of 560 (b)	> the WSF of 320 (b)	> the WSF of 100 (c)

N = 10 fish per concentration.

a Bioassay as conducted at 15 (+ 2°C), mean fish wet weight and standard length,

0.80 (± 0.29) g and 40 (+ 5) mm

b Insufficient mortality for the calculation of an LC5 0.

c There was insufficient mortahty (< 37% to > 63 %) for the calculation of an LC50 by the LC50 computer program (6, 7), therefore, this value is the study director's estimate based on the mortality data.

The 96-hour NOEC was < the WSF of 100 mg/L, based on the mortality and abnormal effects seen at this concentration.

Validity criteria fulfilled:

not specified

Conclusions:

The nominal test concentrations for this study were the water soluble fractions of 100, 180, 320, 560, and 1000 mg/L solutions. The 96-hour LC50 for rainbow trout exposed to the test material was estimated to be > the WSF of 100 mg/L. The NOEC was < the WSF of 100 mg/L.
There appeared to be a compound related lowering of dissolved oxygen.

Executive summary:

The static acute toxicity of the test material to rainbow trout (Oncorhynchus mykiss) was assessed using the methods outlined by the protocol for this study. Water quality parameters of temperature, dissolved oxygen and pH were measured throughout the test and were within acceptable limits in the control test vessels. Culture and acclimation records indicated the fish were in good condition for testing.

The study was conducted at the following nominal concentrations of TegMeR® 804: the WSFs of 100, 180, 320, 560, and 1000 mg/L. The WSF of each nominal test concentration was reached by adding the appropriate weights of compound to 15 L of dilution water, then stirring overnight. The solutions were allowed to stand undisturbed for approximately 6 hours before the solutions were siphoned into the test jars. An effort was made to siphon all but the undissolved compound, which was left remaining in the jar in which it was stirred. The fish were added after the solutions had come to test temperature. All test solutions were noted as being clear at 0 and 24 hours. At 48 and 72 hours however, all solutions were clear except the 100 mg/L B replicate and the A and B replicates of the 320 mg/L test solutions, where a slight amount of compound was seen on the surface. Also at 48 and 72 hours, the 560 mg/L B and the 1000 mg/L A replicates were noted as having oily droplets present on the surface. At 96 hours, there were oily droplets noted on the surface of the 100 mg/L B, 560 mg/L A, and 1000 mg/L B test solutions. All other solutions were clear 96 hours. Ten fish, with a mean wet weight of 0.80 (+ 0.29) g and a mean standard length of 40 (+ 5) mm, were exposed to each test concentration and control.

The 96-hour no-observed effect concentration (NOEC) was < the WSF of 100 mg/L, based on the abnormal effects and/or mortality at this and all higher concentrations. The abnormal effects of mortality, surfacing, loss of equihbrium, dark discoloration, fish on the bottom of the test chamber, labored respiration, erratic swimming, and/or quiescence were observed during the 96 -hour exposure period in the 100, 180, 320, 560, and 1000 mg/L WSF test concentrations.

Description of key information

In a key study on the test substance, the nominal test concentrations for this study were the water soluble fractions of 100, 180, 320, 56, 0, and 1000 mg/L solutions. The 96-hour LC50 for rainbow trout (Oncorhynchus mykiss) exposed to the test substance was estimated to be > the WSF of 100 mg/L. The NOEC was < the WSF of 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The test substance is a member of a category of aliphatic esters submitted by The American Chemistry Council's (ACC) Aliphatic Esters Panel (Panel) under the High Production Volume (HPV) Chemical Challenge Program (ACC, 2003). Information from a Screening Information Data Set (SIDS) developed in that program serves as supplementary or weight-of-evidence in the current dossier. Category ester compounds with similar structures and functionalities were of low acute toxicity to freshwater fish at or above the effective water solubility limits of the substances. The test material decanoic acid, mixed diesters with octanoic acid and triethylene glycol (CAS 68583 -52 -8), had a 96-hour median lethal concentration (LC50) of 430 mg/L. 2,2'-Ethylenedioxydiethyl bis(2 -ethylhexanoate) (CAS 94 -28 -0) caused no mortality to Pimephales promelus in 96 h at the nomial test concentration of 97 mg/L. Ethane-1,2 -diylbis(oxyethane-2,1 -diyl) bisheptanoate (CAS 7434 -40 -4) had a 48 h LC50 value in excess of a nominal 1000 mg/L test concentration. Heptanoic acid, oxybis(2,1 -ethanediyloxy-2,1 -ethanediyl)ester (CAS 70729 -68 -9) had an estimated 48 -h LC50 of 720 mg/L in Golden orfe, at or above the aqueous solubiilty of the test substance. Heptanoic acid, ester with 2,2,4 -trimethyl-1,3 -pentanediol (CAS 71839 -38 -8) was not toxic to Oncorhynchus mykiss in a 96 -h test at a nominal 1000 mg/L.