4-(1,1,3,3-tetramethylbutyl)phenol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

In accordance with Regulation (EC) 1907/2006 Annex XI (1.5) and the relevant ECHA guidance documents, the substances detailed in the table below are grouped for the purposes of read across to reduce the need for unnecessary repeat testing on the basis tha

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.26 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.21 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.39 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

no reference substance tested

Reported statistics and error estimates:

no statisitcs performed, graphical evaluation of the LC50 values

Sublethal observations / clinical signs:

concentration[mg/L]	No. of fishat start of study	observation period
		24 h		48 h		72 h		96 h
		No. dead	mortality [%]	No. dead	mortality[%]	No. dead	mortality[%]	No. dead	mortality[%]
control	10	0	0	0	0	0	0	0	0
control with ethanol	10	0	0	0	0	0	0	0	0
0.21	10	0	0	0	0	0	0	0	0
0.29	10	0	0	3	30	3	30	8	80
0.39	10	7	70	10	100	10	100	10	100
0.60	10	10	100	10	100	10	100	10	100
1.02	10	10	100	10	100	10	100	10	100

Validity criteria fulfilled:

yes

Conclusions:

The read across for 4-tert-octylphenol (CAS: 140-66-9); is based upon the analogous substances to which basic form, degree of substitution of functional groups is not considered to effect the proposed read across for the endpoint of acute aquatic toxicity to fish. Based on the information available for the read across substances, the substance is expected to be acutely toxic to fish.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

11/05/1991-07/06/1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

study has been conducted according to valid study guidelines. Deviations are in the number of fish per concentration and in the absent analytical confirmation of nominal concentrations. This is not expected to compromise the quality of the study results.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Screening study only

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

Method of preparation: preliminary solution in 10% tween 80 - ethanol
Media renewal: daily

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Parkwood Trout Farm, Kent, U.K.
- Length at study initiation: 4.2 ± 0.3 cm
- Weight at study initiation: 1.04 ± 0.32 g Loading: 0.52 g bodyweight/litre


ACCLIMATION
- Health during acclimation: Zero mortalities in the 7 days prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

Approximately 50 mg/l as caCO3

Test temperature:

14°C

pH:

no data

Dissolved oxygen:

Aeration: yes

Salinity:

fresh water test

Nominal and measured concentrations:

Range Finding Study: 0.1, 1.0 and 10 mg/l
Definitive Study: 0.10 mg/l
only nominal concentrations reported

Details on test conditions:

Test water: dechlorinated tap water (sodium thiosulphate)
Medication: none
Test vessels: 20 l

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Range finding study

dates: 22 -26 May 1991

Concentration (mg/l)	Cumulative Mortality (initial population = 2)
	24 Hours	48 Hours	72 Hours	96 Hours
Control	0	0	0	0
0.10	0	0	0	0
1.0	1	2	2	2
10	2	2	2	2

Definitve study

dates: 3 -7 June 1991

Concentration (mg/l)	Cumulative Mortality (initial population = 10)
	24 Hours	48 Hours	72 Hours	96 Hours
Control	0	0	0	0
Solvent control	0	0	0	0
0.10 R1	0	0	0	0
0.10 R2	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The subsatnce is toxic to fish upon short-term exposure

Executive summary:

A study was performed to assess the acute aquatic toxicity of the test material phenol, 4 -(1,1,3,3 -tetramethylbutyl). The method followed the OECD Guidelines for Testing of Chemicals No. 203 "Fish, Acute Toxicity Test. Deviations are in the number of fish per concentration and in the absent analytical confirmation of nominal concentrations (this is not expected to compromise the quality of the study results).

The screening 96 h acute toxicity study was conducted with rainbow trout at nominal test concentrations of 0.10, 1.0 and 10 mg/l. The number of fish was 2 per test concentration in a preliminary study. In the final study, the number of fish was 10 in two replicates at 0.10 mg/l. In parallel a control and a solvent control group were tested. Mortality occurred at 1.0 and 10 mg/l in the range finding study (2 fish). The LC50 at 96 hours exposure was determined to be >0.10 mg/l.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

not provided

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Klimisch 2e. Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: dissolved in 100% ethanol
- Controls: 100% ethanol vehicle control (<0.3% v/v)

Test organisms (species):

Fundulus heteroclitus

Details on test organisms:

TEST ORGANISM
- Common name: killifish
- Source: lab-bred fish originally collected from New River inlet Beaufort, North Carolina, USA. Eggs and milt stripped from adults, combined for 20 min and rinsed in 1% H2O2 to prevent fungal growth for 1 min. Resulting embyros maintained in artificial sea water
- Age at study initiation (mean and range, SD): newly hatched
- Feeding during test: live brine shrimp twice a day

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20 +/- 1 °C

Nominal and measured concentrations:

Nominal concentrations; 10, 25, 50, 100 and 200 µM

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 ml pyrex beaker
- Material, size, headspace, fill volume: 150 ml artificial sea water + 150 µL stock solution
- Renewal rate of test solution: treatment solution renewed daily
- No. of organisms per vessel: 5 larva
- No. of vessels per concentration (replicates): 5

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: artificial sea water

EFFECT PARAMETERS MEASURED:
- Survival: larvae were scored daily for lethality and behavioural abnormalities such as lethargy or lack of feeding

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

2.16 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.29

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.42 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.20

Details on results:

- Other adverse effects: at 2 µM 4-t-OP larvae were observed to display lethargic swimming behaviours and were often unable to feed successfully

Reported statistics and error estimates:

LC50 values determined using PROBIT.
ANOVA procedures used to identified chemical related effects.

Validity criteria fulfilled:

yes

Conclusions:

Acute larval exposures revealed that both NP and 4-t-OP were more lethal to larvae than to embyos. Two week old larvae appeared most sensitive (embryo, newly hatched, 2 week old and 4 week old larvae were tested).
The test substances caused development toxicity in the embryos and larva of F. heteroclitus at concentrations reported in natural waters.

Executive summary:

Kelly and Di Giulio (2000). Klimisch 2e.

In a 96-h acute toxicity study, Killifish (Fundulus heteroclitus) embryos were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 2.1, 5.2, 10.3, and 20.6 mg/L 4-tert-octylphenol under static-renewal conditions.  Killifish (Fundulus heteroclitus) newly hatched, two-week old, and four-week old larvae were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 0.1, 0.2, 0.4, 0.8 mg/L under static-renewal conditions The 96-h LC₅₀for the test with embryos was 3.86 mg/L and for the test with newly hatched, two-week old, and four-week old larvae 0.293, 0.285, and 0.343 mg/L 4-octylphenol, respectively. Results were presented in µM and converted to mg/L here.  

  

This toxicity study is classified as acceptable with restrictions and satisfies the general guideline requirement for a short-term toxicity study.

 

Results Synopsis

 

Test organism size/age: embryo, newly hatched, two-week old, and four-week old larvae

Test Type: static renewal

 

LC₅₀: 

embryo: 18.7 µM or 3.86 mg/L

newly hatched larvae: 1.42µM or 0.293 mg/L

 two-week old larvae: 1.38µM or 0.285 mg/L

four-week old larvae: 1.66 µM or 0.343 mg/L            

Kelly and Di Giulio (2000) was chosen as a supporting study because it was determined to be classified as acceptable with restrictions as it is a saltwater test. This study demonstrates differential sensitivity among life stages with newly hatched and young larvae more sensitive than the embryo stage.  This study shows the most sensitive life stage of F. heteroclitus to acute effects of octylphenol is similar to the findings reported in the Key Study Huels (1993).

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

not provided

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Klimisch 2e. Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: dissolved in 100% ethanol
- Controls: 100% ethanol vehicle control (<0.3% v/v)

Test organisms (species):

Fundulus heteroclitus

Details on test organisms:

TEST ORGANISM
- Common name: killifish
- Source: lab-bred fish originally collected from New River inlet Beaufort, North Carolina, USA. Eggs and milt stripped from adults, combined for 20 min and rinsed in 1% H2O2 to prevent fungal growth for 1 min
- Age at study initiation (mean and range, SD): developed to 4-cell stage

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Observations made for 25 days in total.

Test temperature:

20 +/- 1 °C

Nominal and measured concentrations:

Nominal concentrations; 10, 25, 50 and 100 µM

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic tissue culture dishes
- Material, size, headspace, fill volume: 2 ml artificial sea water + 2 µL stock solution
- Renewal rate of test solution: treatment solution renewed every 24 h for days 1-2 and by artificial sea water days 4-10
- No. of organisms per vessel: 10 embryos
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: artificial sea water

EFFECT PARAMETERS MEASURED:
- Survival: embryos were scored for embryonic differentiation and organogenesis for 10 days postfertilisation. Lethality and specific developmental abnormalities were monitored (craniofacial, cardiac, torso abdominal, tail vascular). Hatch success was measured on day 21 and larval survival recorded at conclusion of the experiment at day 25.

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

18.7 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 1.4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

18.7 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 1.4

Details on results:

- Mortality of control: 0% for 0.3% v/v ethanol control
- Other adverse effects: at 2 µM 4-t-OP larvae were observed to display lerthargic swimming behaviours and were often unable to feed successfully

Reported statistics and error estimates:

LC50 values determined using PROBIT.
Nonparametric (Kruskal-Wallis) procedures used to identified chemical related effects.

Validity criteria fulfilled:

yes

Conclusions:

Complete mortality occurred within the first 48 h among embryos exposed to either NP or 4-t-OP at concentrations of 50 and 100 µM.
The test substances caused development toxicity in the embryos and larva of F. heteroclitus at concentrations reported in natural waters.

Executive summary:

Kelly and Di Giulio (2000). Klimisch 2e.

In a 96-h acute toxicity study, Killifish (Fundulus heteroclitus) embryos were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 2.1, 5.2, 10.3, and 20.6 mg/L 4-tert-octylphenol under static-renewal conditions.  Killifish (Fundulus heteroclitus) newly hatched, two-week old, and four-week old larvae were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 0.1, 0.2, 0.4, 0.8 mg/L under static-renewal conditions The 96-h LC₅₀for the test with embryos was 3.86 mg/L and for the test with newly hatched, two-week old, and four-week old larvae 0.293, 0.285, and 0.343 mg/L 4-octylphenol, respectively. Results were presented in µM and converted to mg/L here.  

  

This toxicity study is classified as acceptable with restrictions and satisfies the general guideline requirement for a short-term toxicity study.

 

Results Synopsis

 

Test organism size/age: embryo, newly hatched, two-week old, and four-week old larvae

Test Type: static renewal

 

LC₅₀: 

embryo: 18.7 µM or 3.86 mg/L

newly hatched larvae: 1.42 µM or 0.293 mg/L

 two-week old larvae: 1.38 µM or 0.285 mg/L

four-week old larvae: 1.66 µM or 0.343 mg/L            

Kelly and Di Giulio (2000) was chosen as a supporting study because it was determined to be classified as acceptable with restrictions as it is a saltwater test. This study demonstrates differential sensitivity among life stages with newly hatched and young larvae more sensitive than the embryo stage.  This study shows the most sensitive life stage of F. heteroclitus to acute effects of octylphenol is similar to the findings reported in the Key Study Huels (1993).

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

not provided

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Klimisch 2e. Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: dissolved in 100% ethanol
- Controls: 100% ethanol vehicle control (<0.3% v/v)

Test organisms (species):

Fundulus heteroclitus

Details on test organisms:

TEST ORGANISM
- Common name: killifish
- Source: lab-bred fish originally collected from New River inlet Beaufort, North Carolina, USA. Eggs and milt stripped from adults, combined for 20 min and rinsed in 1% H2O2 to prevent fungal growth for 1 min. Resulting embyros maintained in artificial sea water
- Age at study initiation (mean and range, SD): 4 weeks
- Feeding during test: live brine shrimp twice a day

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20 +/- 1 °C

Nominal and measured concentrations:

Nominal concentrations; 10, 25, 50, 100 and 200 µM

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 ml pyrex beaker
- Material, size, headspace, fill volume: 150 ml artificial sea water + 150 µL stock solution
- Renewal rate of test solution: treatment solution renewed daily
- No. of organisms per vessel: 5 larva
- No. of vessels per concentration (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: artificial sea water

EFFECT PARAMETERS MEASURED:
- Survival: larvae were scored daily for lethality and behavioural abnormalities such as lethargy or lack of feeding

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

2.14 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.28

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.66 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.18

Details on results:

- Other adverse effects: at 2 µM 4-t-OP larvae were observed to display lethargic swimming behaviours and were often unable to feed successfully

Reported statistics and error estimates:

LC50 values determined using PROBIT.
ANOVA procedures used to identified chemical related effects.

Validity criteria fulfilled:

yes

Conclusions:

Acute larval exposures revealed that both NP and 4-t-OP were more lethal to larvae than to embyos. Two week old larvae appeared most sensitive (embryo, newly hatched, 2 week old and 4 week old larvae were tested).
The test substances caused development toxicity in the embryos and larva of F. heteroclitus at concentrations reported in natural waters.

Executive summary:

Kelly and Di Giulio (2000). Klimisch 2e.

In a 96-h acute toxicity study, Killifish (Fundulus heteroclitus) embryos were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 2.1, 5.2, 10.3, and 20.6 mg/L 4-tert-octylphenol under static-renewal conditions.  Killifish (Fundulus heteroclitus) newly hatched, two-week old, and four-week old larvae were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 0.1, 0.2, 0.4, 0.8 mg/L under static-renewal conditions The 96-h LC₅₀for the test with embryos was 3.86 mg/L and for the test with newly hatched, two-week old, and four-week old larvae 0.293, 0.285, and 0.343 mg/L 4-octylphenol, respectively. Results were presented in µM and converted to mg/L here.  

  

This toxicity study is classified as acceptable with restrictions and satisfies the general guideline requirement for a short-term toxicity study.

 

Results Synopsis

 

Test organism size/age: embryo, newly hatched, two-week old, and four-week old larvae

Test Type: static renewal

 

LC₅₀: 

embryo: 18.7 µM or 3.86 mg/L

newly hatched larvae: 1.42 µM or 0.293 mg/L

 two-week old larvae: 1.38 µM or 0.285 mg/L

four-week old larvae: 1.66 µM or 0.343 mg/L            

Kelly and Di Giulio (2000) was chosen as a supporting study because it was determined to be classified as acceptable with restrictions as it is a saltwater test. This study demonstrates differential sensitivity among life stages with newly hatched and young larvae more sensitive than the embryo stage.  This study shows the most sensitive life stage of F. heteroclitus to acute effects of octylphenol is similar to the findings reported in the Key Study Huels (1993).

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

not provided

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Klimisch 2e. Meets generally accepted scientific standards, well documented and acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: dissolved in 100% ethanol
- Controls: 100% ethanol vehicle control (<0.3% v/v)

Test organisms (species):

Fundulus heteroclitus

Details on test organisms:

TEST ORGANISM
- Common name: killifish
- Source: lab-bred fish originally collected from New River inlet Beaufort, North Carolina, USA. Eggs and milt stripped from adults, combined for 20 min and rinsed in 1% H2O2 to prevent fungal growth for 1 min. Resulting embyros maintained in artificial sea water
- Age at study initiation (mean and range, SD): 2 weeks
- Feeding during test: live brine shrimp twice a day

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20 +/- 1 degree celsius

Nominal and measured concentrations:

Nominal concentrations; 10, 25, 50, 100 and 200 uM

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 ml pyrex beaker
- Material, size, headspace, fill volume: 150 ml artificial sea water + 150 microL stock solution
- Renewal rate of test solution: treatment solution renewed daily
- No. of organisms per vessel: 5 larva
- No. of vessels per concentration (replicates): 5

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: artificial sea water

EFFECT PARAMETERS MEASURED:
- Survival: larvae were scored daily for lethality and behavioural abnormalities such as lethargy or lack of feeding

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

1.76 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.20

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.38 µmol/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: standard error of the mean 0.00

Details on results:

- Other adverse effects: at 2 uM 4-t-OP larvae were observed to display lerthargic swimming behaviours and were often unable to feed successfully

Reported statistics and error estimates:

LC50 values determined using PROBIT.
ANOVA procedures used to identified chemical related effects.

Validity criteria fulfilled:

yes

Conclusions:

Acute larval exposures revealed that both NP and 4-t-OP were more lethal to larvae than to embyos. Two week old larvae appeared most sensitive (embryo, newly hatched, 2 week old and 4 week old larvae were tested).
The test substances caused development toxicity in the embryos and larva of F. heteroclitus at concentrations reported in natural waters.

Executive summary:

Kelly and Di Giulio (2000). Klimisch 2e.

In a 96-h acute toxicity study, Killifish (Fundulus heteroclitus) embryos were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 2.1, 5.2, 10.3, and 20.6 mg/L 4-tert-octylphenol under static-renewal conditions.  Killifish (Fundulus heteroclitus) newly hatched, two-week old, and four-week old larvae were exposed to 4-tert-octylphenol at nominal concentrations of 0 (control, solvent control), and 0.1, 0.2, 0.4, 0.8 mg/L under static-renewal conditions The 96-h LC₅₀for the test with embryos was 3.86 mg/L and for the test with newly hatched, two-week old, and four-week old larvae 0.293, 0.285, and 0.343 mg/L 4-octylphenol, respectively. Results were presented in µM and converted to mg/L here.  

  

This toxicity study is classified as acceptable with restrictions and satisfies the general guideline requirement for a short-term toxicity study.

 

Results Synopsis

 

Test organism size/age: embryo, newly hatched, two-week old, and four-week old larvae

Test Type: static renewal

 

LC₅₀: 

embryo: 18.7 µM or 3.86 mg/L

newly hatched larvae: 1.42µM or 0.293 mg/L

 two-week old larvae: 1.38µM or 0.285 mg/L

four-week old larvae: 1.66 µM or 0.343 mg/L            

Kelly and Di Giulio (2000) was chosen as a supporting study because it was determined to be classified as acceptable with restrictions as it is a saltwater test. This study demonstrates differential sensitivity among life stages with newly hatched and young larvae more sensitive than the embryo stage.  This study shows the most sensitive life stage of F. heteroclitus to acute effects of octylphenol is similar to the findings reported in the Key Study Huels (1993).

Reference 7

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1992-01-13 to 1992-01-17

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0 - 1.5 mg/l
- Sampling method: not mentioned
- Sample storage conditions before analysis: not mentioned

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 70%

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

TEST ORGANISM
- Common name: Golden Orfe
- Strain: Leuciscus idus melanotus Heckel
- Source: Fischzucht Eggers, Hohenwestedt, Germany
- Age at study initiation (mean and range, SD): not mentioned
- Length at study initiation (length definition, mean, range and SD): 6 ± 2 cm, no further details mentioned
- Weight at study initiation (mean and range, SD): not mentioned
- Method of breeding: not mentioned
- Feeding during test: none

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): The fish were kept in 200 l earthenware basins in flow rate.
- Type and amount of food: TetraMin, administration in an amount of approx. 3 % of body weight
- Feeding frequency: daily
- Health during acclimation (any mortality observed): The animals showed a normal behaviour pattern at the beginning of the test and were free from visible diseases.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

12.2 °dH

Test temperature:

20 ± 1 °C

pH:

7.5 - 7.9

Dissolved oxygen:

8.2 - 8.9 mg/l

Salinity:

not mentioned

Nominal and measured concentrations:

Nominal: 0.3, 0.45, 0.7, 1.0 and 1.5 mg/l
Measured: 0.3, 0.4, 0.54, 0.83 and 1.42 mg/l, 48.5% decrease after 24 h therefore concentrations for calculation: 0.21, 0.29, 0.39, 0.6 and 1.02 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): not mentioned
- Material, size, headspace, fill volume: 20 l
- Aeration: continuously, no further details mentioned
- Renewal rate of test solution (frequency/flow rate): daily renewal
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: not mentioned

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking water, Gelsenwasser AG, Germany
no further details mentioned
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours darkness
- Light intensity: not mentioned

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality after 24, 48, 72 and 96 hours; pH, temperature, O2 saturation at the end of the test

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not mentioned
- Range finding study: not mentioned

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.26 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.21 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.39 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

no reference substance tested

Reported statistics and error estimates:

no statisitcs performed, graphical evaluation of the LC50 values

Sublethal observations / clinical signs:

concentration[mg/L]	No. of fishat start of study	observation period
		24 h		48 h		72 h		96 h
		No. dead	mortality [%]	No. dead	mortality[%]	No. dead	mortality[%]	No. dead	mortality[%]
control	10	0	0	0	0	0	0	0	0
control with ethanol	10	0	0	0	0	0	0	0	0
0.21	10	0	0	0	0	0	0	0	0
0.29	10	0	0	3	30	3	30	8	80
0.39	10	7	70	10	100	10	100	10	100
0.60	10	10	100	10	100	10	100	10	100
1.02	10	10	100	10	100	10	100	10	100

Validity criteria fulfilled:

yes

Conclusions:

No study conclusion is given by the authors.

Executive summary:

In a 96-h acute toxicity study, Golden Orfe (Leusiscus. Idus)were exposed to octylphenol at measured concentrations of 0 (control, solvent control), 0.3, 0.4, 0.54, 0.83, and 1.42 mg/L under static-renewal conditions.  There was a 48% decrease in concentration in test vessels after 24 h, therefore, concentrations used in endpoint calculation were 0.21, 0.29, 0.39, 0.6, and 1.02 mg/L octylphenol. The 96-h LC₅₀was 0.26 mg/L.  Based on the results of this study, octylphenol would be classified as toxic to Leusiscus. Idus.

  

This toxicity study is classified as acceptable and satisfies the guideline requirement for a short-term toxicity study.

 

Results Synopsis

 

Test organism size/age (mean length): 6+/-2 cm

Test Type: static renewal

 

LC₅₀:  0.26 mg /L                        

 

Huels, (1993) was chosen as a Key study because it is a GLP Guideline study and followed OECD guidelines. Results provided in this study indicate the sensitivity of L idus is similar to that of other fish species evaluated including those that are considered sensitive to toxicant such as the fathead minnow at 96 h LC₅₀0.29 mg/L (unpublished report) and the rainbow trout at 96 h LC₅₀>0.1 mg/L (Sewel 1991), all of which were more sensitive to octylphenol than reported for Fundulus heteroclitus by Kelly (2000). 

Reference 8

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1984

Reliability:

2 (reliable with restrictions)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Method is not explicitly stated, however it matches the detail of OECD 203 in terms of execution.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

Stock solution: 14,550 mg/l in acetone

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead Minnows
- Source: ABC in-house culture
- Length at study initiation (length definition, mean, range and SD): 23 mm +/- 1.9 mm
- Weight at study initiation (mean and range, SD): 0.20 g +/- 0.05 g
- Feeding during test: No

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: Rangen's standard commercial fish food
- Feeding frequency: Daily

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

225 - 275 ppm Calcium Carbonate

Test temperature:

15 - 20 degrees centigrade

pH:

7.8 - 8.3

Dissolved oxygen:

9.2 - 10.2 ppm

Nominal and measured concentrations:

Nominal: 0.047, 0.091, 0.18, 0.39, and 0.7 mg/l
Measured: 0.041, 0.077, 0.15, 0.34, and 0.63 mg/l

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 0.25 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % confidence interval

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 0.077 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % confidence interval

Sublethal observations / clinical signs:

Concentration	Fish Mortality (%)
	24 hours	48 hours	72 hours	96 hours
Control (Solvent)	0	0	0	0
0.047 mg/l	0	0	0	0
0.091 mg/l	0	0	0	0
0.18 mg/l	0	0	0	0
0.39 mg/l	65	90	90	90
0.70 mg/l	100	100	100	100

Validity criteria fulfilled:

yes

Conclusions:

LC50 is 0.25 mg/l, NOEC is 0.077 mg/l.

Executive summary:

A dynamic 96-hour toxicity study was conducted to determine the acute toxicity of octylphenol to fathead minnows (Pimephales promelas). The methods employed are those recommended by the US EPA, and are equivalent to OECD 203. Water quality parameters were measured throughout the study and were within acceptable limits.

A flow-through proportional diluter system was used to deliver a constant concentration of octylphenol into the test aquaria. Nominal exposure concentrations were 0.047, 0.091, 0.18, 0./39 and 0.70 mg/l. The mean measured concentrations were 0.041, 0.077, 0.15, 0.34 and 0.63 mg/l. The mean measured concentrations were used in the LC₅₀ determinations for octylephenol to fathead minnows.

As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A, The estimated 96-hour LC₅₀ (7 x 10^-5 mg/l) and 95% confidence interval (4.2-14 x 10^-5 mg/l) were within the 95% confidence limits reported in the literature, indicating that the fish were in suitable conditions for testing.

The results of the study are summarised below. Based on the lack of mortality and abnormal effects, the results indicate a 96 -hour LC50 and 95% confidence interval of 0.25 (0.15 -0.34) mg/l, and a NOEC of 0.077 mg/l for octylphenol, based on measured concentrations.

Hours of expsoure	LC50	95% confidence interval	NOEC
24	0.29	0.15-0.63	0.15
48	0.25	0.15-0.34	0.077
72	0.25	0.15-0.34	0.077
96	0.25	0.15-0.34	0.077

Description of key information

 96hr LC50 of >0.26 mg/L  for Leusiscus idus(Hϋls 1993) 
 Fundulus heteroclitusto 96 hr LC50 of 0.28 mg octylphenol/L (2 week old larvae)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.26 mg/L

Marine water fish

Marine water fish

Dose descriptor:

LC50

Effect concentration:

0.285 mg/L

Additional information

 A review of toxicity test results with short-term octylphenol exposure to fish resulted in three reliable 96 hr studies that included two freshwater and one saltwater species. Test endpoints were based on survival of exposure and test results were reported as LC₅₀concentrations. Freshwater fish results included a LC₅₀of 0.26 mg octylphenol/L forLeusiscus idus(Hϋls 1993) and LC₅₀of >0.1 for Onchorhynchus mykiss (Safepharm 1991). Kelly et al., (2000) exposed four different early life stages (embryo, newly hatched, 2 week old larvae, and 4 week old larvae) of Fundulus heteroclitusto octylphenol and reported a range of LC₅₀concentrations from 0.28 mg octylphenol/L (2 week old larvae) to 3.86 mg octylphenol/L (embryo).  Kelly et al., (2000) results show decreasing sensitivity of life stages for Fundulus heteroclitus to be 2 week old larve > newly hatches larvae > 4 week old larvae > embryo.  

 

The Hϋls (1993) study was selected as key studies because it waswell documented and provided standard survival endpoints for the freshwater test species. The supporting Safepharm (1991) study provides similar documentation, however it is only a screening study, therefore is not selected as a key study. The Kelly et al, (2000) study results provide a range of LC₅₀concentrations for the more sensitive life stages of Fundulus heteroclitus that agree well with the findings of the key study by Hϋls 1993.