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

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

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 that the substances are similar on the basis of a common functional groups.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

20 d

Dose descriptor:

NOEC

Effect conc.:

0.042 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

20 d

Dose descriptor:

LOEC

Effect conc.:

0.091 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

parent generation

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.009 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.028 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

53 d

Dose descriptor:

NOEC

Effect conc.:

0.02 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parents animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.016 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.215 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.116 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.215 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.04 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parents animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.06 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parents animals

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

0.001 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.025 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parents animals

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parents animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.013 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.025 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

0.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.024 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.039 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

mean length of parent

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.13 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.03 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

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 chronic aquatic toxicity to invertebrates. Based on the information available for the read across substances, the substance is expected to be toxic to invertebrates following long term exposure.

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. Test concentrations were not measured and some testing information is not provided.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

not specified

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

none noted

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution prepared by dissolving p-nonylphenol with solvent and diluted
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylsulfoxide (DMSO)
- Concentration of solvent in control and test solutions: 50 mg/L (preliminary experiment showed this concentration did not have adverse effects on daphnids

Test organisms (species):

other: Daphnia galeata

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia galeata
- Source: laboratory culture, original source collected from LAke Sagmiko Japan. Culture maintained as 12 isofemale clones for at least 4 generations before testing.
- Feeding during test: yes
- Food type: Chlorella vulgaris
- Amount: 15000000 cells/ml
- Frequency: every 2 or 3 days

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
Five to ten neonates were isolated to start new generation.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

30 mg/L CaCO3

Test temperature:

20 °C for cultures

pH:

6.9-7.2 s.u.

Dissolved oxygen:

6.8-7.0 mg/L for cultures

Salinity:

NA

Nominal and measured concentrations:

nominal concentrations: 0, 3, 10, 30, 50, 70, 100 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 200 ml vessel
- Renewal rate of test solution (frequency/flow rate): every 2 or 3 days
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated/carbon filtered tap water
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned
-Age of test organisms: < 24 h old at test initiation

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D for cultures


VEHICLE CONTROL PERFORMED: yes

Survival and reproduction assessed at renewals.
Test continued to test second generation by arbitrarily sampling from first or second broods of parent generation. Second generation organisms were exposed in the same way as first generation.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Control survival acceptable as determined by graphs presented in paper.
No adverse effect seen on survival or reproduction in first or second generation below 50 µg/L nonylphenol.

Reported statistics and error estimates:

Randomization test (Manly 1991) was used to determine statistical pairwise comparisons between treatments and control for longevity and fecundity. The intrinsic rate of natural increase for the life table data was fitted to the Euler-Lotka equation.

Validity criteria fulfilled:

not specified

Conclusions:

The study author concludes that the acute LC50 (0.061 mg/L) is a good estimator of chronic population-level effects of nonylphenol.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia galeata was studied under static renewal conditions. Daphnids were exposed to control and nonylphenol at nominal concentrations of 0.003, 0.010, 0.030, 0.05, 0.070, 0.10 mg/Lnonylphenol.  The 21-day NOEC based on survival and reproduction was 0.050 mg/L nonylphenol.  The sublethal effects included were fecundity and survival of second generation. 

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flow through Static, Static Renewal):static renewal

 

NOEC (survival): 0.05 mg/L nonylphenol

NOEC (reproduction): 0.05 mg/L nonylphenol

 

Tanaka and Nakanishi (2002) was selected as a supporting study because it provides toxicity information on the sensitivity of the sensitive test organism Daphnia galeata to nonylphenol. Results of this study are consistent with chronic toxicity test performed on the same genus of Daphnia (Brooke 1993, Sun and Gu 2005, and Comber et al. 1993).

Reference 3

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. Test concentrations were not measured and some testing information is not provided.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

not specified

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

none noted

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution prepared, dilution water added after volitilization of solvent by blowing nitrogen for acute tests
- Controls: yes dilution water control only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): hexane but noted for acute test only. Not clear if solvent used in chronic test. Study author notes that the content of hexane in test medium could be ignored due to volatilization in acute tests.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: laboratory culture
- Feeding during test: yes
- Food type: Chlorella
- Amount: 800000 cells/ml
- Frequency: daily

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
Lab cultures were maintained at 20 °C under 2000 Lux intensity with 16L:8D and fed with green algae, Chlorella. 6-24 h old daphnids used for testing.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

2.5 mM Ca+Mg

Test temperature:

20 ± 1 °C for cultures

pH:

7.5-7.8 s.u.

Dissolved oxygen:

not given

Salinity:

NA

Nominal and measured concentrations:

nominal concentrations based on 24 h LC50 test results.
Test concentrations noted as 1/32, 1/16, 1/8, 1/4, 1/2 of the 24 h LC50 (0.37 mg/L)
which were 0.013, 0.025, 0.05, 0.1, 0.2 mg/L nonylphenol

Details on test conditions:

TEST SYSTEM
- Test vessel: 150 ml vessel
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 3
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16L:8D for cultures
- Light intensity: 2000 lux

VEHICLE CONTROL PERFORMED: uncertain

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.025 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.013 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.025 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

100% survival in controls

Reported statistics and error estimates:

ANOVA used to analyze data to detect significant difference between control and treatments. Dunnett's Procedure followed. p=<0.05

Results for Survival, Reproduction, and Interval of Molting of Daphnia magna Exposed to NP

NP (mg/L)	Survival (%)	# of Offspring/adult	Interval of molting (d)
control	100	131	2.47
0.013	100	118	2.42
0.025	88.9	109*	2.41
0.05	77.8*	94*	2.32*
0.10	55.6*	77*	2.25*
0.20	33.3*	55*	2.12*

*significantly different from control

Validity criteria fulfilled:

yes

Conclusions:

The study author concludes that NP can have an chronic effects on molting process, reproduction and survival of Daphnia magna. Study also concluded that hardness, pH, and humic acid can effect the toxicity of NP in acute tests.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia manga was studied under static renewal conditions. Daphnids were exposed to control and nonylphenol at nominal concentrations of 0.013, 0.025, 0.05, 0.1, 0.2 mg/L.  The 21-day NOEC based on reproduction was 0.013 mg/L nonylphenol and survival was 0.025 mg/L nonylphenol.  The sublethal effects included were number of offspring/surviving parent.  Production of offsprings in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.025 mg /L.  The most sensitive end point was reproduction.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flowthrough Static, Static Renewal):static renewal

 

NOEC (survival): 0.025 mg/L nonylphenol

NOEC (reproduction): 0.013 mg/L nonylphenol

 

Sun and Gu (2005) was selected as a supporting study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study are consistent with test results of Brooke (1993), and Comber et al. (1993) for Daphnia magna.

Reference 4

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guidelines study with acceptable restrictions which do not impair the overall conclusion for the data. Concentrations were not measured

Qualifier:

no guideline available

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

NA

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: dissolved in solvent then ultrapure distilled water, stock then added in appropriate amounts to filter-sterilized seawater to reach test concentrations
- Controls: solvent and dilution water controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethyl sulfoxide (DMSO)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)):0.001%

Test organisms (species):

other aquatic crustacea: Tigriopus japonicus

Details on test organisms:

TEST ORGANISM
- Common name: copepod
- Source: in-house cultures, original source originated from pond of Fisheries Labortory at the University of Tokyo, near Lake Hamana
- Feeding during test: yes
- Food type: Nanochloropsis oculata
- Amount: 700000 cells/ml
- Frequency: daily at renewal
-age of test organisms: <24 h nauplii

ACCLIMATION
- Acclimation period: none as cultured in lab
- Acclimation conditions (same as test or not): same
- Type and amount of food: Nanochloropsis oculata

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

not mentioned

Test temperature:

25 ± 1 °C

pH:

not provided

Dissolved oxygen:

not provided

Salinity:

25 ppt

Nominal and measured concentrations:

nominal: 0.01, 0.1, 1.0, 10 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: well polystyrene plates
- Material, size, fill volume: 1 ml in well plates days 1-7, 10 ml in well plates days 8-21
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 3
- Biomass loading rate: not mentioned

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filter-sterilized seawater diluted with ultrapure distilled water
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Photoperiod: 12L:12D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): survival assessed daily, fecundity, sex ratio, nauplii stage duration and maturation stage durage were also assessed for both parent and progeny.

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

parent generation

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

There were no statistical differences in survival from treatments and controls for parent or progeny. There were also no significant differences in fecundity (of F1 generation) or sex ratio (for F0 and F1 generation). However, a significant difference was seen in fecundity in the 1 µg/L concentration where reproduction was actually higher in the treatment than in the control. Only survival and reproduction results are considered for assessment since they are traditional endpoints.

Reported statistics and error estimates:

Dunnett's was used to determine significance between number of days to reach copepodid stage and sexual maturity. Fecundity data were assessed using ANOVA and Dunnett's test. p=<0.05

Summary of Toxicity Results of Exposure of Tigriopus japonicus to Octylphenol for 21 Days (Parent Generation)

	Nauplii Stage ( avg days)	Maturation (avg days)	Fecundity	Sex Ratio (F:M)	Survival (%)
Control	5.6	13.6	28.1	0.6	93.3
Solvent Control	5.5	13.1	34.9	0.7	88.3
0.01 (ug/L)	5.4	13.1	33.3	0.9	93.6
0.1 (ug/L)	6.3*	13.6	38.4	0.7	90.0
1 (ug/L)	6.0*	13.4	40.1*	0.4	98.3
10 (ug/L)	5.6	13.2	34.6	0.5	92.8

Summary of Toxicity Results of Exposure of Tigriopus japonicus to Octylphenol for 21 Days (F1 Generation)

	Nauplii Stage ( avg days)	Maturation (avg days)	Fecundity	Sex Ratio (F:M)	Survival (%)
Control	5.7	13.1	11.2	0.8	93.6
Solvent Control	5.8	13.3	15.4	0.5	95.3
0.01 (ug/L)	6.5*	13.8	20.7	0.7	93.6
0.1 (ug/L)	5.9	13.8	16.6	0.7	83.7
1 (ug/L)	6.7*	14.2*	15.3	0.7	98.3
10 (ug/L)	6.0*	14.6*	13.2	1.6	94.1

*Significant difference p<0.05

Validity criteria fulfilled:

not applicable

Conclusions:

The study authors conclude that results indicate that endocrine disruption could occur in copepods when exposed to environmentally relevant concentrations of octylphenol and other compound tested (4-nonylphenol, 17B estradiol, bisphenol A).

Executive summary:

The 21-day-chronic toxicity of octylphenol to Tigriopus japonicus was studied under static renewal conditions. Copepods were exposed to control, solvent control, and test chemical at concentrations of 0.00001, 0.0001, 0.001, and 0.01 mg octylphenol./L.  The 21-day NOEC based on {sublethal effect; reproduction} was 0.01 mg octylphenol./L for both the parent and prodgeny generation.  The sublethal effects included were reproduction as well as sex ratio, maturation and days in nauplii stage.  Production of offsprings in the treated groups indicated that octylphenol did not have an effect on the reproduction at concentrations greater than 0.01 mg/L octylphenol.  The most sensitive end point was days in nauplii stage for the F1 generation.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic toxicity study with saltwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): <24hrs

Test Type (Flowthrough Static, Static Renewal): semi-static

 

NOEL:  >0.01 mg/L octylphenol

               Endpoint(s) Effected:  reproduction

 

Marcial et al (2003) was selected as a supporting study because results from this study showed toxicity results for the sensitive test organism, Tigriopus japonicus. No other reliable long-term data was available for the toxicity of octylphenol to saltwater invertebrates.

 

Reference 5

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guidelines study with acceptable restrictions which do not impair the overall conclusion for the data. Concentrations were not measured.

Qualifier:

no guideline available

GLP compliance:

not specified

Analytical monitoring:

no

Details on sampling:

NA

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: dissolved in solvent then ultrapure distilled water, stock then added in appropriate amounts to filter-sterilized seawater to reach test concentrations
- Controls: solvent and dilution water controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethyl sulfoxide (DMSO)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.001%

Test organisms (species):

other aquatic crustacea: Tigriopus japonicus

Details on test organisms:

TEST ORGANISM
- Common name: copepod
- Source: in-house cultures, original source originated from pond of Fisheries Labortory at the University of Tokyo, near Lake Hamana
- Feeding during test: yes
- Food type: Nanochloropsis oculata
- Amount: 700000 cells/ml
- Frequency: daily at renewal
- Age of test organisms: <24 h nauplii

ACCLIMATION
- Acclimation period: none as cultured in lab
- Acclimation conditions (same as test or not): same
- Type and amount of food: Nanochloropsis oculata

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

NA

Test temperature:

25 ± 1 °C

pH:

not provided

Dissolved oxygen:

not provided

Salinity:

25 ppt

Nominal and measured concentrations:

nominal: 0.01, 0.1, 1.0, 10 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: well polystyrene plates
- Material, size, fill volume: 1 ml in well plates days 1-7, 10 ml in well plates days 8-21
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 3
- Biomass loading rate: not mentioned

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filter-sterilized seawater diluted with ultrapure distilled water
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Photoperiod: 12L:12D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : survival assessed daily, fecundity, sex ratio, nauplii stage duration and maturation stage durage were also assessed for both parent and progeny.

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

parent generation

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

There were no statistical differences in survival from treatments and controls for parent or progeny. There were also no significant differences in fecundity or sex ratio for F0 and F1 generations. However, a significant difference was seen in days in nauplii stage and days from birth to sexual maturation for both parent and F1 generation. Only survival and reproduction results are considered for assessment since they are traditional endpoints.

Reported statistics and error estimates:

Dunnett's was used to determine significance between number of days to reach copepodid stage and sexual maturity. Fecundity data were assessed using ANOVA and Dunnett's test. p=<0.05

Summary of Toxicity Results of Exposure of Tigriopus japonicus to Nonylphenol for 21 Days (Parent Generation)

	Nauplii Stage ( avg days)	Maturation (avg days)	Fecundity	Sex Ratio (F:M)	Survival (%)
Control	5.6	13.6	28.1	0.6	93.3
Solvent Control	5.5	13.1	34.9	0.7	88.3
0.01 (ug/L)	5.4	13.4	35.7	1.2	91.7
0.1 (ug/L)	5.7	13.5	35.7	1.0	93.4
1 (ug/L)	6.4	14.0	31.8	0.9	89.4
10 (ug/L)	6.8	14.8*	28.5	0.7	93.0

Summary of Toxicity Results of Exposure of Tigriopus japonicus to Nonylphenol for 21 Days (F1 Generation)

	Nauplii Stage ( avg days)	Maturation (avg days)	Fecundity	Sex Ratio (F:M)	Survival (%)
Control	5.7	13.1	11.2	0.8	93.6
Solvent Control	5.8	13.3	15.4	0.5	95.3
0.01 (ug/L)	5.9	13.6	13.4	0.7	84.3
0.1 (ug/L)	6.1*	13.8	15.3	0.9	90.2
1 (ug/L)	6.3*	14.1*	15.9	0.6	88.1
10 (ug/L)	6.3*	14.3*	13.5	0.5	91.7

*Significant difference p<0.05

Validity criteria fulfilled:

not applicable

Conclusions:

The study authors conclude that results indicate that endocrine disruption could occur in copepods when exposed to environmentally relevant concentrations of nonylphenol and other compound tested (octylphenol, 17B estradiol, bisphenol A).

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Tigriopus japonicus was studied under static renewal conditions. Copepods were exposed to control, solvent control, and test chemical at concentrations of 0.00001, 0.0001, 0.001, and 0.01 mg nonylphenol./L.  The 21-day NOEC based on {sublethal effect; reproduction} was 0.01 mg nonylphenol./L for both the parent and progeny generation.  The sublethal effects included were reproduction as well as sex ratio, maturation and days in nauplii stage.  Production of offspring in the treated groups indicated that nonylphenol did not have an effect on the reproduction at concentrations greater than 0.01 mg/L octylphenol.  The most sensitive end point was days in nauplii stage for the F1 generation.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic toxicity study with saltwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): <24 h

Test Type (Flow through Static, Static Renewal): semi-static

 

NOEL:  >0.01 mg/L nonylphenol

               Endpoint(s) Effected:  reproduction

 

Marcial et al (2003) was selected as a supporting study because results from this study showed toxicity results for the sensitive test organism, Tigriopus japonicus. No other reliable long-term data was available for the toxicity of nonylphenol to this species. However, results are in agreement with results for other saltwater invertebrates, Americamysis bahia (Kuhn et al. 2001) and Tisbe battagliai (Bechmann 1999).

 

Reference 6

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

February 27, 1997 - March 1997, April 1997-June 1997

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study meets generally accepted scientific standards and is described in sufficient detail.

Qualifier:

according to guideline

Guideline:

other: ASTM 1990. Standard Guide for Conducting Life-cycle Toxicity Tests with Saltwater Mysids. E1191-90

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

yes

Details on sampling:

sampled collected weekly

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: yes dilution water control and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): mixture of acetone and triethylene glycol (TEG)

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

TEST ORGANISM
- Common name: mysid shrimp
- Source: cultures maintained at the Atlantic Ecology Division (Narragansett, RI, USA)
- Feeding during test: yes
- Food type: Artemia nauplii
- Amount: 150 48 h post hatch nauplii
- Frequency: daily

ACCLIMATION
- Acclimation period: none needed as test organisms cultured in lab

Test type:

flow-through

Water media type:

saltwater

Limit test:

no

Total exposure duration:

28 d

Post exposure observation period:

none

Hardness:

NA

Test temperature:

25 °C

pH:

7.6-8.1s.u.

Dissolved oxygen:

5.7-7.0 mg/L

Salinity:

30 ppt

Nominal and measured concentrations:

nominal concentrations: 0, 0, 3.125, 6.25, 12.5, 25, 50, 100, 200 µg/L
average measured: ND, ND, 5.94, 7.94, 9.46, 15.28, 21.06, 27.56, 49.24 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 µm mesh chambers set in glass tanks
- Type of flow-through (e.g. peristaltic or proportional diluter): proportional diluter
- Renewal rate of test solution (frequency/flow rate): 45 ml/min
- No. of organisms per vessel: 15 per chamber, two chambers per replicate side, 60 total organisms per treatment
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filtered Narragansett Bay seawater
- Culture medium different from test medium: unknown
- Intervals of water quality measurement: twice a week
- Age of test organisms: 24 h old
- Total organic carbon: below detection limits
- Photoperiod: 4L:10D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): survival and reproduction assessed daily, growth assessed at test termination

VEHICLE CONTOL PERFORMED: yes

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.009 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.028 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

94% survival in controls
76% survival in solvent control

Solvent control used to determine significant differences

Reported statistics and error estimates:

ANOVA used to analyze data to detect significant difference between control and treatments. p=<0.05

Results for Survival, Reproduction, and Growth of Americamysis bahia Exposed to NP

Average Measured NP (mg/L)	Average Survival (%)	Average weight/female (mg)	Average # young/female reproductive days (AFRD) (arsine transformed)	Time to First Brood (days)
control	94	0.11	0.94	14
solvent control	76	0.10	0.68	14
0.00594	95	0.08	0.49	15
0.00794	99	0.10	0.40	16
0.00946	99	0.09	0.14	17
0.01528	91	0.09	0.07*	25
0.02106	100	0.07	0*	no young
0.02756	79	0.05*	0*	no young
0.04924	25*	0.06*	0*	no young

*significantly different from control

Validity criteria fulfilled:

not specified

Conclusions:

The toxicity tests (life-cycle and multi-generational) were performed to validate a population prediction model. The study author concludes that population model was able to project within a few micrograms per liter the concentration effects would be seen.

Executive summary:

The 28-day-chronic toxicity of nonylphenol to the mysid shrimp, Americamysis bahia, was studied under flow-through conditions. A. bahia were exposed to control, solvent control, and nonylphenol at measured concentrations of 5.94, 7.94, 9.46, 15.28, 21.06, 27.56, 49.24 µg/L nonylphenol.  The 28-day NOEC based on reproduction was 0.00946 mg/L nonylphenol and survival was 0.02756 mg/L nonylphenol.  The sublethal effects included were growth and average number of offspring per available female.  Production of offspring in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.00946 mg/L.  The most sensitive end point was reproduction.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with marine invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): 24 h

Test Type (Flowthrough Static, Static Renewal): flow through

 

NOEC (survival): 0.02756 mg/L nonylphenol

NOEC (reproduction): 0.00946 mg/L nonylphenol

 

Kuhn et al. (2001) was selected as a supporting study because it provides toxicity information on the sensitivity of a common marine test organism Americamysis bahia to nonylphenol.  Results of this study show the relative sensitivity of marine invertebrates may be similar or more sensitive than freshwater invertebrates such as Daphnia magna.

Reference 7

Endpoint:

long-term toxicity to aquatic invertebrates

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: Meets generally accepted scientific standards, well documented and acceptable for assessment. Study did not document dilution water characteristics and had some analytical variability.

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD 219

Deviations:

yes

Remarks:

water only exposure

Principles of method if other than guideline:

Method followed cited as Benoit et al. (1997). Chironomus tentans life cycle test: Design and evaluation for use in assessing toxicity of contaminated sediments. Environ. Toxicol. Chem Vol 16 pp1165-1176.

Water only exposure

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

yes

Details on sampling:

Samples collected on test days 0, 4, 11, 18, 25, 42, 48, 53.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): not mentioned

Stock solutions were prepared by adding to flask and diluted with Lake Superior water. Stock was stirred at moderate rate

Test organisms (species):

other: Chironomus tentans

Details on test organisms:

TEST ORGANISM
- Common name: Chironomid
- Source: on-site culture
- Details on collection: egg masses collected 72 h prior to test initiation
- Age of animals at beginning of exposure: < 24 h old larvae
- Feeding during test: yes
- Food type: Tetrafin food slurry
- Amount: 1.5 ml/ test chamber
- Frequency: daily

ACCLIMATION
- Acclimation period: none needed as cultured in lab
- Acclimation conditions (same as test or not): same dilution water

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

53 d

Post exposure observation period:

none

Hardness:

not provided

Test temperature:

23 +/- 1 °C

pH:

7.73 +/- 0.13 s.u.

Dissolved oxygen:

7.05+/- 0.59 mg/L

Salinity:

NA

Nominal and measured concentrations:

nominal: 0, 12.5, 25, 50, 100, 200 µg/L
Actual measured concentrations not provided. Test concentrations remained with 20% of nominal for first 18 days, then decreased by 50% or less of target value.

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 300 ml beaker
- Sediment volume: 5 ml of 146 µm washed sand

EXPOSURE REGIME
- No. of organisms per container (treatment): 12
- No. of replicates per treatment group: 4 for growth and survival, 6 for reproduction
- Feeding regime: daily
- Type and preparation of food: Tetrafin slurry
- Amount of food: 1.5 ml

RENEWAL OF OVERLYING WATER
- Details on volume additions: complete renewal
- Flow-rate: 0.5 tank turnovers per day

OVERLYING WATER CHARACTERISTCS
- Type of water (e.g. deionized, ground water, sea water, Elendt medium acc. to OECD 219): lake Superior water
- Source of water (if non-standard medium) Lake Superior
- Total organic carbon: not provided

Unknown if substrate is natural or artificial washed sand

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): daily, growth determined on Day 20


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 0.5

Duration:

20 d

Dose descriptor:

NOEC

Effect conc.:

0.042 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

20 d

Dose descriptor:

LOEC

Effect conc.:

0.091 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

It was noted that measured test concentrations were within 20% of nominal for 18 days. By day 25, test concentrations had decreased to 50% or less of nominal without reason. A 50% difference in toxicant concentration was also seen from tanks versus exposure beakers.

Reported statistics and error estimates:

ANOVA followed by Tukey's pairwise comparisons p<0.05.

Summary of Results for Toxicity of Nonylphenol to Chironomus tentans

Treatment (µg/L)	Survival Day 20 (%)	Growth Day 20 (mg/organism)	Survival from Day 20 to end of test (%)	Sex ratio (M/F)	Fecundity (eggs/female)	Viability (% hatch)
control	73	0.52	96	0.35	753	85
12.5	71	0.55	87	0.64	695	88
25	73	0.65	89	0.33	702	85
50	81	0.57	75	0.40	647	86
100	77	0.64	82	0.47	847	77
200	38*	0.83	92	0.56	684	91

*Significantly different p<0.05

Validity criteria fulfilled:

not specified

Conclusions:

Study was performed to confirm observations seen in a field experiment. Results indicate that although survival of C. tentans is affected by nonylphenol (LOEC 0.091 mg/L), ultimate reproductive success in surviving organisms is not affected, at the concentrations tested.

Executive summary:

The 20-day-chronic toxicity of nonylphenol to Chironomus tentans was studied under flow through conditions. Chironomids were exposed to control, and test chemical at nominal concentrations of 12.5, 25, 50, 100, and 200 µg./L nonylphenol.  The 20 day NOEC based on survival was 0.042 mg /L nonylphenol.  The sublethal effects assessed included growth, fecundity, and percent hatch of next generation.  Due to analytical issues, calculated point estimates are not reliable for sublethal effects after Day 20 of test.  The most sensitive end point was survival.

 

This study is classified as acceptable.

 

Results Synopsis

 

Test Organism Age (e.g. 1^st instar): first instar

Test Type (Flowthrough Static, Static Renewal): flow through

 

NOEC (survival): 0.042 mg/L nonylphenol

Kahl et al. (1997) was chosen as a supporting study because results indicate that Chironomus tentans could be a more sensitive species to nonylphenol toxicity. Due to the analytical issues occurring during the test and lack of some experimental information (dilution water characteristics) this study was not deemed acceptable for key study determination. However, this study can be used to support other findings for long-term toxicity of nonylphenol to aquatic invertebrates.

Reference 8

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

March 2004 - April 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study was performed according to Guidelines with acceptable restrictions which do not impair the overall conclusion from the data. Test concentrations were not measured.

Qualifier:

according to guideline

Guideline:

other: ISO/CD 20665 procedure (2001)

Deviations:

not specified

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

no

Details on sampling:

none

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: original stock culture obtained from MicroBio Tests Inc., testing organisms cultured in-house
- Feeding during test: yes
- Food type: algae, P. subcapitata
- Frequency: daily

ACCLIMATION
- Acclimation period: none since cultured in lab


METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: < 24 h old magna used for testing were obtained from in house cultures maintained in ISO hard medium, temperature of 20+/-2 °C, under 300-450 lux

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Post exposure observation period:

none

Hardness:

250 +/- 20 mg/L CaCO3

Test temperature:

25 °C

pH:

8.0+/-0.2 s.u.

Dissolved oxygen:

at air saturation, test was aerated at start of test

Salinity:

NA

Nominal and measured concentrations:

specific concentrations not given. There were 7 concentrations using a two-fold dilution series. Author noted in communications that test concentrations stared from 20 times less than acute EC50 value: from 0.01 to 0.00015 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers
- Material, size, headspace, fill volume: 20 ml of test solution
- Renewal rate of test solution (frequency/flow rate):every 24 hrs
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates):10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted "hard" water as per USEPA
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Photoperiod: 16L:8D
- Light intensity: 500 lux
- Test organisms fed daily 100 ul suspension of algae P. subcapitata, fish food and yeast.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :survival and reproduction at every renewal

VEHICLE CONTROL PERFORMED: yes

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

0.001 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

7 day EC50 given as 0.008 mg/L NP
Control mortality < 20%

Reported statistics and error estimates:

Analysis of results used Toxcalc (1996). EC50 calculated using Maximum Likelihood-Logit method.

A total of 8 akylphenols were tested. Each compound was tested three times. Results given are average of three tests.

Validity criteria fulfilled:

yes

Conclusions:

The study author concludes that exposure to the eight alkylphenols tested causes acute and chronic effects.

Executive summary:

The 7-day-chronic toxicity of nonylphenol to Ceriodaphnia dubia was studied under static renewal conditions. Daphnids were exposed to control, solvent control, and nonylphenol at nominal concentrations.  The 7-day NOEC based on reproduction was 0.001 mg/L nonylphenol.  The sublethal effect included number of offspring/surviving parent.  Production of offspring in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.001 mg /L.  The most sensitive end point was reproduction. Survival NOEC was not provided, but 7 -day EC50 was 0.008 mg/L nonylphenol.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flow through Static, Static Renewal):static renewal

 

NOEL:  0.001 mg/L (reproduction)

 

Isidori et al. (2006) was selected as a supporting study because it provides toxicity information on the sensitivity of the test organism Ceriodaphnia dubia to nonylphenol. Results of this study indicate that Ceriodaphnia may be more sensitive than Daphnia magna to nonylphenol. Results of this test are not in agreement with England (1995) Study #41756. However, study author notes that NOEC results are not precise since the results are not calculated and subject to study design.

Reference 9

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1992-05-06 to 1992-05-27

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: OECD 202, part II, 1984

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

not mentioned

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1 g test substance was dissolved in 1 L acetone
- Controls: yes, test water and vehicle control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): stock solution: 100 %; final test concentration: max 100 µg/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Straus, IRCHA
- Source: laboratory breed
- Age of parental stock (mean and range, SD): not older than 24 hours
- Feeding during test
- Food type: Scenedesmus subspicatus
- Amount: day 0-6: 0.4 x 10E7 cells/daphnia; day 7-21: 2.0 x 10E7 cells/daphnia
- Frequency: daily

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
The females were maintained in tap water. A reddish dark-brown coloration of the animals indicated that they were healthy. At two to three day intervals, the exuviate were siphoned off and the water was changed. In the course of this, the offspring was removed from the breeding vessels. At regular intervals of approx. 4 - 5 weeks, offspring was isolated from the vessels. They served as a basis for further breeding.
Females were separated from the offspring 24 hours before the start of the test. The offspring produced overnight were transferred into the test water and were used for the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

not mentioned

Test temperature:

20 ± 1 °C

pH:

7.7 - 8.6, not adjusted, the increase of pH did not affect the reproduction rate

Dissolved oxygen:

95 - 114% saturation

Salinity:

synthetic fresh water, Elendt (1990)

Nominal and measured concentrations:

Up to 3 µg/L the concentrations were not reproducible. From 10 µg/L the measured concentrations of the freshly prepared solutions without dapnia lay within the permitted deviations (< 20%) from the nominal concentrations and were stable during the course of the study. A reduction of the measured values in the test solutions were considered to be caused by absorption to algae and daphnia. All values were calculated on the basis of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Type (delete if not applicable): open
- Material, size, fill volume: glass, 250 mL, 200 mL (day 0- 6), 80 mL on days 7 to 21
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): 3 times per week (Mo, We, Fr)
- No. of organisms per vessel: 5 up to day 6, 1 on days 7 to 21
- No. of vessels per concentration (replicates): 4 up to day 6, max. 10 from day 7 to 21
- No. of vessels per control (replicates): 4 up to day 6, max. 10 from day 7 to 21
- No. of vessels per vehicle control (replicates): 4 up to day 6, max. 10 from day 7 to 21
- Biomass loading rate: not mentioned

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic fresh water, Elendt (1990)
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 20/4 h (light/dark)
- Light intensity: 1500 lux

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Details on results:

- Mortality of parent animals: control = 0%, solvent = 10%, 0.3 µg/L = 0%, 1.0 µg/L = 10%, 3.0 µg/L = 20%, 10 µg/L = 0%, 30 µ/L = 10%, 100 µg/L = 0%
- Number of alive young daphnia/parent animal on day 21 (reproduction rate): control = 74.6 ± 30.7; solvent = 77 ± 24.8; 0.3 µg/L = 73.7 ± 20.8; 1.0 µg/L = 72.4 ± 18.1; 3.0 µg/L = 83.8 ± 14.5; 10 µg/L = 78.9 ± 19.8; 30 µg/L = 69.7 ± 25.7; 100 µg/L = 69.4 ± 20.1

Reported statistics and error estimates:

t-test (alpha 0.05): all concentrations not significantly different

Validity criteria fulfilled:

yes

Conclusions:

None provided by study authors

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia magna was studied under static renewal conditions. Daphnids were exposed to control, solvent control, and nonylphenol at average measured concentrations.  The 21-day NOEC based on reproduction was >0.1 mg/L nonylphenol and survival was >0.1 mg/L nonylphenol.  The sublethal effects included were number of offspring/surviving parent. 

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flowthrough Static, Static Renewal):static renewal

 

NOEC:  >0.1 mg/L (reproduction)

NOEC:>0.1 (survival)

 

Huls (1992) was selected as a supporting study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study are consistent with test results of Brooke (1993), and Sun.and Gu (2005) for Daphnia magna.

Reference 10

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

November 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study meets generally accepted scientific standards and is acceptable for assessment.

Qualifier:

according to guideline

Guideline:

other: ISO10872:2010

Deviations:

not specified

Principles of method if other than guideline:

Study references Traunspurger et al. 1997 as method followed. The method presented by Traunspurger et al. 1997 has since been approved as a standard test method ISO 10872:2010 which is a 96 h test. This study performed test for 72 h, but was considered by the authors to be a full life-cycle. Although was performed as water only exposure to a sediment dwelling organism, USEPA has accepted water-only studies on sediment organisms (Hylella azeteca).

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

yes

Details on sampling:

Test concentrations measured at beginning and end of test.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water

Test organisms (species):

other aquatic worm: Caenorhabditis elegans

Details on test organisms:

TEST ORGANISM
- Common name: nematode
- Source: cultured in lab
- Age of animals at beginning of exposure: juvenile, first stage mean body length = 270+/- 16 µm
- Feeding during test: yes
- Food type: bacteria
- Amount: 1000000000 cells/ml

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Remarks on exposure duration:

full life cycle

Post exposure observation period:

none

Hardness:

not provided

Test temperature:

20 °C

pH:

7.2 s.u.

Dissolved oxygen:

not given

Salinity:

NA

Nominal and measured concentrations:

nominal: 50, 100, 150, 200, 250, 300, 350 µg/L
average measured: 40.2, 65.6, 106.5, 150.8, 189.2, 213.9, 235.2 µg/L

Details on test conditions:

TEST SYSTEM
- Test container (material, size): polystyrene multiwells

EXPOSURE REGIME
- No. of organisms per container (treatment):5
- No. of replicates per treatment group:7
- No. of replicates per control / vehicle control:7
- Feeding regime: at test initiation
- Type and preparation of food: bacteria (E. coli)


OVERLYING WATER CHARACTERISTCS
- Type of water (e.g. deionized, ground water, sea water, Elendt medium acc. to OECD 219): Study states NP solution was added to phosphate buffer. M9 buffer consisted of 5g/L NaCl, 6 g/L Na2HPO4, 3 g/L KH2PO4, 0.25 g/L MgSO4 7H2O


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : growth and reproduction at test end

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 0.235 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

100% survival in control and all treatments
Growth and reproduction wre enhanced in the presence of NP.

Reported statistics and error estimates:

ANOVA used to determine significance between control and treatments

Validity criteria fulfilled:

not specified

Remarks:

missing water quality characteristics

Conclusions:

Study authors conclude that results from this study showed that NP can enhance growth and reproduction of Caenorhabditis elegans at environmentally relevant concentrations in polluted sediments. These results could be ecologicallly relevant as this species plays a significant role in the benthic food web.

Executive summary:

The 72 h-chronic toxicity of nonylphenol to the nematode, Caenorhabditis elegans, was studied under static conditions. Nematodes were exposed to control, and nonylphenol at average measured concentrations of 40.2, 65.6, 106.5, 150.8, 189.2, 213.9, and 235.2 µg./L nonylphenol.  The 72 -h NOEC based on survival, reproduction, and growth was > 0.2352 mg /L nonylphenol for all three effects. Growth and reproduction was enhanced at higher concentrations of NP.

The sublethal effects assessed included reproduction and growth.  The most sensitive end point was not able to be determined.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): juvenile

Test Type (Flow through Static, Static Renewal): static

 

72 -hr NOEC (survival, reproduction and growth): >0.2352 mg/L nonylphenol

Hoss et al. (2002) was chosen as a supporting study because provides toxicity results of nonylphenol to the sediment organism, Caenorhabditis elegans. Although this study was performed in a water only exposure, it can be considered reliable enough to use as supporting the key study in the toxicity of nonylphenol to aquatic organisms.

Reference 11

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

February 1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study was performeed according to Guidelines with acceptable restrictions which do not impair the overall conclusion from the data. Test concentrations were not measured and study does not indicate it was performed according to GLP.

Qualifier:

according to guideline

Guideline:

other: USEPA 600/4-89/001

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: Standard Methods 1980 15th ed

Qualifier:

according to guideline

Guideline:

other: USEPA 660/3-75-009 1975

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone


4.0 mg/L stock solution was prepared by in acetone.

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: original stock culture obtained from Aquatic Biosystems, Inc July 1990, testing organisms cultured in-house
- Feeding during test: yes
- Food type: algae, trout chow, and yeast suspension
- Frequency: daily

ACCLIMATION
- Acclimation period: none since cultured in lab


METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: < 24 h old magna used for testing were obtained adult daphnids that had been isolated the previous day. Adults had been cultured for 28 days previously and had not signs of stress or disease.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Post exposure observation period:

none

Hardness:

170-174 mg/L CaCO3

Test temperature:

25 °C

pH:

8.4-8.6 s.u.

Dissolved oxygen:

not given

Salinity:

NA

Nominal and measured concentrations:

nominal: control, solvent control, 0.003, 0.01, 0.03, 0.10, 0.30 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 30 ml plastic cups
- Material, size, headspace, fill volume: 15 ml of test solution
- Renewal rate of test solution (frequency/flow rate): every 24 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates):10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted "hard" water as per USEPA
- Culture medium different from test medium: no
- TOC = <1.0 mg/L
- Chemical parameters (pH, temperature, hardness, alkalinity, dissolved oxygen and conductivity) measured daily

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D
- Light intensity: 50-70 footcandles


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : at every renewal

VEHICLE CONTROL PERFORMED: yes

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

0.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Concentration response for survival was erratic and therefore, is not the most reliable indicator of toxicity. Study author states that reproductive results are more reliable indication of chronic effects. Also, there appears to be a delay in first brood for the higher concentrations confirming that reproduction may be more indicative of chronic effects.

Reported statistics and error estimates:

NOEC and LOEC were determined by using SAS. Control treatments were compared using chi-square and Fisher's exact test to determine acceptability for pooling. Dunnett's test was used to compare mean survival to controls as well as for reproduction. ANOVA was used to determine statistical differences between mean survival/reproduction among treatments.

a 7-day LC50 was calculated using program developed by Stephan et al.

Nominal Conc. (mg/L)	% Survival	Mean No. of live offspring/surviving parent/day	Days to First Brood
Control	90	5.1	3.3
Solvent control	100	4.3	3.2
0.003	80	5.6	3.4
0.01	60*	4.2	3.3
0.03	70	4.6	3.3
0.10	50*	3.6	3.0
0.30	100	3	4.8

*Significantly different from pooled controls

Validity criteria fulfilled:

yes

Conclusions:

The study author concludes that reproduction may be a better indicator of chronic effects of nonylphenol than survival due to delayed first brood and erratic survival results.

Executive summary:

The 7-day-chronic toxicity of nonylphenol to Ceriodaphnia dubia was studied under static renewal conditions. Daphnids were exposed to {control, solvent control, and nonylphenol) at nominal concentrations of 0.003, 0.01, 0.03, 0.10, 0.30 mg/L.  The 7-day NOEC based on reproduction was 0.10 mg/L nonylphenol.  The sublethal effect included number of offspring/surviving parent.  Production of offspring in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.1 mg /L.  The most sensitive end point was reproduction. Survival NOEC was 0.01 mg/L however, this value was determined to be unreliable due to erratic survival response.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24hrs

Test Type (Flow through Static, Static Renewal):static renewal

 

NOEL:  0.1 mg/L (reproduction)

LOEC: 0.3 mg/L

 

England (1995) was selected as a supporting study because it provides toxicity information on the sensitivity of the test organism Ceriodaphnia dubia to nonylphenol. Results of this study indicate that Ceriodaphnia may be less sensitive than Daphnia magna to nonylphenol. Results of this test are in agreement with England (1995) Study #41756.

Reference 12

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991-10-02 to 1991-12-18

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: OECD 202, part II, 1984

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

not mentioned

Vehicle:

yes

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: Straus, IRCHA
- Justification for species other than prescribed by test guideline:
- Source: laboratory breed
- Age of parental stock (mean and range, SD): not older than 24 hours
- Feeding during test
- Food type: Scenedesmus subspicatus
- Amount: day 0-6: 0.4 E7 cells/daphnia; day 7-21: 2.0 E7 cells/daphnia
- Frequency: daily

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
The females were maintained in tap water. A reddish dark-brown coloration of the animals indicated that they were healthy. At two to three day intervals, the exuviate were siphoned off and the water was changed. In the course of this, the offspring was removed from the breeding vessels. At regular intervals of approx. 4 - 5 weeks, offspring was isolated from the vessels. They served as a basis for further breeding.
Females were separated from the offspring 24 hours before the start of the test. The offspring produced overnight were transferred into the test water and were used for the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

not mentioned

Test temperature:

20 ± 1 °C

pH:

7.3 - 8.2, not adjusted

Dissolved oxygen:

83 - 105% saturation

Salinity:

synthetic fresh water, Elendt

Nominal and measured concentrations:

The test substance concentrations were stable during the course of the study. The measured concentrations lay within the permitted deviations (< 20%) from the nominal concentrations. Therefore all values were calculated on the basis of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Type (delete if not applicable): open
- Material, size, fill volume: glass, 250 ml, 200 ml (day 0- 6) and 150 ml, 80 ml on day 7 until 21
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): 3 times per week (Mo, We, Fr)
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 4
- Biomass loading rate: not mentioned

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M4 medium, Elendt
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 20/4 h (light/dark
- Light intensity: 1500 lux

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.03 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: control = 0%, solvent = 0%, 0.01 mg/l = 10%, 0.03 mg/l = 0%, 0.1 mg/l = 20%, 0.3 mg/l = 0%, 1.0 mg/l = 100%
- Total number of alive young daphnia on day 21: control = 118, solvent = 107, 0.01 mg/l = 107, 0.03 mg/l = 106, 0.1 mg/l = 99, 0.3 mg/l = 88, 1.0 mg/l = 0

Reported statistics and error estimates:

t-test (alpha 0.05): 0.01, 0.03 and 0.1 mg/l not significant; 0.3 and 1.0 mg/l significant

Validity criteria fulfilled:

yes

Conclusions:

No conclusion available from study authors.

Executive summary:

The 21-day-chronic toxicity of octylphenol to Daphnia magna was studied under static renewal conditions. Daphnids were exposed to control, solvent control, and test chemical at concentrations of 0.01, 0.03, 0.1, 0.3 and 1.0 mg octylphenol./L.  The 21-day NOEC based on {sublethal effect; reproduction} was 0.03 mg octylphenol./L.  The sublethal effects included were reproduction.  Production of offspring in the treated groups indicated that octylphenol had or an effect on the reproduction at concentrations greater than 0.03 mg/L octylphenol.  The most sensitive end point was reproduction.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): <24hrs

Test Type (Flow through Static, Static Renewal): semi-static

 

NOEL:  0.03 mg/L octylphenol

LOEC:  0.1 mg/L octylphenol                   Endpoint(s) Effected:  reproduction

 

Huls (1992) was selected as a key study because results from this study showed toxicity results for the preferred sensitive test organism, Daphnia magna. No other reliable long-term data was available for the toxicity of octylphenol to freshwater invertebrates.

 

Reference 13

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

not provided

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to generally accepted scientific stndards and is described in sufficient detail.

Qualifier:

according to guideline

Guideline:

other: OECD 202

Deviations:

yes

Remarks:

modified by individually separating test organisms

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations new test solutions were measured at each renewal, old solutions were renewed on the following day.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 90 µg/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): not mentioned

stock solutions were prepared by dissolving weighed amounts of test chemical into acetone.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: D. magna Straus, clone type 5
- Source: original stock from Institute national de recherche chimique applique IRCHA and subsequently cultured at ICI Group Environmental Laboratory
- Feeding during test: yes
- Food type: algae and yeast suspension
- Frequency: daily

ACCLIMATION
- Acclimation period: none since cultured in lab


METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: < 24 h old magna used for testing were obtained from a single culture vessel

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

180 +/- 20 mg/L CaCO3

Test temperature:

20 °C

pH:

8.25+/- 0.25 s.u.

Dissolved oxygen:

not given

Salinity:

NA

Nominal and measured concentrations:

nominal: control, solvent control, 0.018, 0.032, 0.056, 0.1, 0.18, 0.32 mg/L
average measured: ND,ND, 0.014, 0.024, 0.039, 0.071, 0.13, 0.25 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessels
- Material, size, headspace, fill volume: 80 ml of test solution
- Renewal rate of test solution (frequency/flow rate): every 48 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates):10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted "hard" water as per USEPA
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : at every renewal

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
48 h acute study performed
- Test concentrations: control, solvent control, 0.032, 0.056, 0.1, 0.18, 0.32, 0.56 mg/L
48 h EC50 = 0.19 mg/L

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.024 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.039 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

mean length of parent

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.13 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Reported statistics and error estimates:

Effects on parent survival calculated using contingency table (exact test) procedure. Dunnett's procedure following one way analysis of variance used to assess mean numbers of total and live offspring produced by each parent.

Nominal Conc. (mg/L)	Mean Meas. Conc. (mg/L)	% Parent Survival	Mean No. of live offspring/surviving parent	Mean length of suriving parent (mm)
Control	<0.001	100	205	4.72
Solvent control	<0.001	100	201	4.71
0.018	0.014	90	188	4.65
0.032	0.024	90	197	4.62
0.056	0.039	100	169	4.67
0.10	0.071	70	162	4.57
0.18	0.13	40	140	4.48
0.32	0.25	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The relationship between the acute and chronic toxicity studies generated in this study is similar to previously reported comparisons of acute and chronic toxicity to D. magna. The author concludes that there is an approximate safety factor of X10 between the NOEC for nonylphenol and highest reported environmental concentrations.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia manga was studied under static renewal conditions. Daphnids were exposed to control, solvent control, and nonylphenol at average measured concentrations of 0.014, 0.024, 0.039, 0.071, 0.13, 0.25 mg/L.  The 21-day NOEC based on reproduction was 0.024 mg/L nonylphenol and growth was 0.039 mg/L nonylphenol.  The sublethal effects included were number of offspring/surviving parent and length of parent.  Production of offspring in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.024 mg /L. The most sensitive end point was reproduction.

 

This study is classified as an acceptable key study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar): < 24 h

Test Type (Flow through Static, Static Renewal):static renewal

 

NOEL:  0.024 mg/L (reproduction)

0.039 mg/L (growth)

0.13 (survival)

 

Comber et al (1993) was selected as a key study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study are consistent with test results of Brooke (1993), and Sun.and Gu (2005) for Daphnia magna.

Reference 14

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

not provided

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to generally accepted scientific standards and is described in sufficient detail. It is not known if study was performed according to GLP.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: ASTM 1991. Standard Guide for Conducting Renewal Life-Cycle Toxicity Tests with Daphnia magna

Deviations:

not specified

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
none

Analytical monitoring:

yes

Details on sampling:

Fresh solutions were sampled on days prepared (Monday, Wednesday, Friday) and old solutions were sampled on days replaced and from pooled replicates. Samples also sampled at test termination.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: no solvents used to generate stock solution. Solution was stirred with magnetic stirrer. Test concentrations were prepared by withdrawling stock solution by pump. Solution was withdrawn near the bottom of the flask.
- Controls: dilution water control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: original stock from Environmental Research Lab - Deluth, MN and cultured at University of Wisconsin-Superior Lab
- Feeding during test: yes
- Food type: algae and yeast suspension
- Frequency: Monday, Wednesday, Friday regime

ACCLIMATION
- Acclimation period: none since cultured in lab


METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: test organisms obtained from 4th brood parents of cultures.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

167 +/- 9.2 mg/L CaCO3

Test temperature:

20.5 +/- 0.5 °C

pH:

8.46+/- 0.11 s.u.

Dissolved oxygen:

8.7 +/-0.5 mg/L

Salinity:

NA

Nominal and measured concentrations:

average measured: ND, 44.3, 63.1, 116, 215, 500 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml plastic cups
- Material, size, headspace, fill volume: 80 ml of test solution
- Renewal rate of test solution (frequency/flow rate): every Monday, Wednesday, Friday
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted "hard" water as per USEPA
- Culture medium different from test medium: no
- Alkalinity: 120 +/- 9.1 mg/L
- Conductivity: 536 +/- 21umhos/cm

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : temp. measured daily, DO measured twice a week, Hardness, alkalinity, pH measured weekly

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.116 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.215 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.116 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

growth

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.215 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Chronic 21-day test on D. magna was performed 3 times. Author notes that Test 1 concentrations declined an average of 48 % between renewals. Test 2 had control failure. Only results from Test 3 were valid and are used for REACH assessment. However, all three test results are in agreement. NOEC for reproduction for Test 1 is 0.0995 mg/L and Test 2 is 0.0773 mg/L nonylphenol.

Reported statistics and error estimates:

ANOVA used to determine NOECs. Dunnett's test used to determine significant differences from controls in reproduction. p= 0.05

Summary of Test 3 Results for Chronic Toxicity of Nonylphenol to Daphnia magna

Mean Meas. Conc. (mg/L)	% Parent Survival	Mean No. of live offspring/surviving parent	Mean length of suriving parent (mm)	Mean No. of young/starting adult
Control	80	111	3.94	94.8
0.0443 +/- 0.0111	90	113	3.93	102
0.0631 +/- 0.0093	100	103	3.84	103
0.116 +/- 0.120	90	118	3.84	116
0.215 +/- 0.017	60	87.5	3.62*	53.3*
0.500 +/- 0	0*	0*	0*	0*

*Significant difference from control at p=0.05

Validity criteria fulfilled:

yes

Conclusions:

The study author does not make specific conclusions but simply reports study results only.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia manga was studied under static renewal conditions. Daphnids were exposed to control and nonylphenol at average measured concentrations of 0.0443, 0.0631, 0.116, 0.215, 0.500 mg/L.  The 21-day NOEC based on reproduction was 0.116 mg/L nonylphenol and growth was 0.116 mg/L nonylphenol.  The 21 -day NOEC based on survival was 0.215 mg/L nonylphenol. The sublethal effects included were number of offspring/surviving parent and length of parent.  Production of offsprings in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.116 mg /L.  The most sensitive end point was reproduction.

 

This study is classified as an acceptable key study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flowthrough Static, Static Renewal): static renewal

 

NOEC: 0.116 mg/L nonylphenol (reproduction)

NOEC: 0.116 mg/L nonylphenol (growth)

NOEC: 0.215 mg/L nonylphenol (survival)

 

Brooke (1993) was selected as a supporting study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study show there is a range of toxicity of nonylphenol to Daphnia magna. Although results from this study are slightly less sensitive the results are in agreement with test results of Comber et al. (1993), and Sun.and Gu (2005) for Daphnia magna.

Reference 15

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. Test concentrations were not measured and some testing information is not provided.

Qualifier:

equivalent or similar to guideline

Guideline:

other: ISO 2000 Water Quality-Determination of Long Term Toxicity of Substances to Daphnia Magna Straus(Cladocera, Crustacea)

Deviations:

not specified

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

none noted

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution prepared, NP dissolved in ethanol
- Controls: yes dilution water control and solvent controls
- 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)): <0.1 mg/L

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: laboratory culture, original stock obtained from USEPA Cincinnati, OH
- Feeding during test: yes
- Food type: Chlorella vulgaris supplemented with Tetrafin fish food
- Frequency: 1400000000 cells of algae and 4 mg/L fish food suspension twice a day

Test organisms were cultured at 20 °C with photoperiod 16L:8D according to ISO 1996
Test organisms were < 24 h old

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

not provided

Test temperature:

20 ± 1 °C for cultures

pH:

not provided

Dissolved oxygen:

not provided

Salinity:

NA

Nominal and measured concentrations:

nominal concentrations solvent control, 0.02, 0.04, 0.06, 0.08, 0.10 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 ml beakers with 40 ml test solution
- Renewal rate of test solution (frequency/flow rate): 3 times a week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted
- Culture medium different from test medium: no
- Intervals of water quality measurement: not known

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D for cultures

VEHICLE CONTROL PERFORMED: uncertain

Survival molting frequency and number of offspring were assessed.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.04 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.06 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

100% survival of control organisms
NOEC values are not specifically presented in the paper but inferred based on information and text provided.
Second generation effects were also assessed and significant fecundity effects were seen as low as 0.04 mg/L nonylphenol. However, testing procedures for second generation are lacking. Results are given for first generation only.

Reported statistics and error estimates:

ANOVA followed by Dunnett's multiple t-test method. p=<0.05

Results for Survival and Reproduction of Daphnia magna Exposed to NP

NP (mg/L)	Adult Mortality	Cumulative moults/female	Cumulative offspring/female
control	0	10.3	68 +/-9
0.02	10	10	70 +/-8
0.04	30	9.9	75 +/-10
0.06	40	10.5	59 +/-7
0.08	80	9.0	48 +/-2*
0.10	100	-	-

*significantly different from control (p=0.05)

Second generation offspring and moults assessed.

Validity criteria fulfilled:

not specified

Conclusions:

The purpose of the study was to determine if selected toxicants had toxic effects on Daphnia magna. It was concluded that nonylphenol does have an effect on survival and reproduction of Daphnia magna.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia manga was studied under static renewal conditions. Daphnids were exposed to solvent control and nonylphenol at nominal concentrations of 0.02, 0.04, 0.06, 0.08, 0.10 mg/L.  The 21-day NOEC based on reproduction was 0.06 mg/L nonylphenol and survival was 0.04 mg/L nonylphenol.  The sublethal effects included were number of offspring/surviving parent and moulting frequency.  Production of offsprings in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.06 mg /L.  The most sensitive end point was survival.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flow through Static, Static Renewal):static renewal

 

NOEC (survival): 0.04 mg/L nonylphenol

NOEC (reproduction): 0.06 mg/L nonylphenol

 

Brennan et al. (2006) was selected as a supporting study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study are consistent with test results of Brooke (1993), Baldwin et al (2006), and the key study, Comber et al. (1993) for Daphnia magna.

Reference 16

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

January 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions.

Qualifier:

no guideline available

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

Test concentrations sampled immediately after preparation and on Day 4

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution prepared by using acetone as solvent. Test solutions prepared by adding stock solution to flask with seawater/algae-mix and stirring with magnetic stirrer for 2 minutes.
- Controls: yes dilution water and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of solvent in test solutions: 25 µl/L

Test organisms (species):

other: Tisbe batagliai

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: laboratory culture
- Feeding during test: yes
- Food type: algae D. tertiolecta and I. galbana cultures
- Amount: 5000 cells/ml D. tertiolecta and 10000 cells/ml I. galbana in 10% of test solutions
- Frequency: at renewal

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
Nauplii obtained from isolated females bearing eggs 24 h before test initiation

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

53 d

Post exposure observation period:

none

Hardness:

NA

Test temperature:

10 °C

pH:

not given

Dissolved oxygen:

not given

Salinity:

25 psu

Nominal and measured concentrations:

nominal: 0, 0, 31, 125, 500 µg/L
average measured: 20, 74, 300 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: plastic cyliders with 50 µm plankton net at the base. Cyliders were placed on a rack in a container with 1.5 L of test medium.
- Renewal rate of test solution (frequency/flow rate): every 4th day
- No. of organisms per vessel: 50
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3

On Day22, female were isolated, one per chamber, for fecundity observation until day 54.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: filtered seawater from Oslofjord, Norway
- Culture medium different from test medium: no
- Intervals of water quality measurement: not mentioned

- Age of test organisms: nauplii, < 24 h

OTHER TEST CONDITIONS
- Photoperiod: 12L:12D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : survival, sexed assessed at Day 26, females assessed daily for egg production from Day 22-53.

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):

no

Duration:

53 d

Dose descriptor:

NOEC

Effect conc.:

0.02 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

TL2 had control failure and is not a valid test. Only TL1 results are presented.

Reported statistics and error estimates:

ANOVA used to analyze data to detect significant difference between control and treatments. p=<0.05

Summary of Results for 53 Day Exposure of Tisbe battagliai to Nonylphenol

	Control	Acetone Control	31 µg/L NP	125 µg/L NP	500 µg/L NP
% Survival	86	85	55	0	0
% Egg-producing Females	82	83	54	-	-
# nauplii / egg producing females (Day 1 -39)	87	93	93	-	-
Realized fecundity with Day 26 eggs	36	38	25	-	-
Realized fecundity Day 1 -54	65	76	45	-	-
Intrinsic rate of natural increase	0.110	0.112	0.101	-	-

Test concentrations above are as nominal.

No statistical significant difference in any life table parameter between controls and 31 µg/L NP treatment.

TL2 experiment tested an additional concentration of 62 µg/L NP. However, control survival was 66% on Day 26 and acetone control survival was 47% on Day 26.

Validity criteria fulfilled:

not applicable

Remarks:

no guidelines available for this test species

Conclusions:

The study author concludes that NP did not significantly affect any demographic parameter of the marine copepod exposed to 31 µg/L NP. The authors state that NP concentrations measured in rivers and estuaries are orders of magnitude lower than the effect concentrations seen in this experiment for T. battagliai.

Executive summary:

The 53-day-chronic toxicity of nonylphenol to the marine copepod, Tisbe battagliai was studied under static renewal conditions. T. battagliai were exposed to control, solvent control and nonylphenol at nominal concentrations of 0.031, 0.125, and 0.500 µg/L mg/L.  The 53-day NOEC based on survival and reproduction was 0.031 mg/L nonylphenol.  The sublethal effect included was fecundity. 

 

This study is classified as an acceptable supporting study for a chronic toxicity study with marine invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flowthrough Static, Static Renewal): static renewal

 

NOEC (survival): 0.031 mg/L nonylphenol

NOEC (reproduction): 0.031 mg/L nonylphenol

 

Bechmann (1999) was selected as a supporting study because it provides toxicity information on the sensitivity of a marine test organism, Tisbe battagliai

to nonylphenol. Results of this study show the relative sensitivity of marine invertebrates may be similar to freshwater invertebrates such as Daphnia magna.

Reference 17

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

unknown

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. Test concentrations were not measured and some testing information is not provided.

Qualifier:

equivalent or similar to guideline

Guideline:

other: ASTM 1988. Standard Guide for Conducting Renewal Life-Cycle Toxicity Tests with Daphnia magna

Deviations:

not specified

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

none noted

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution prepared, NP dissolved in ethanol while stirring.
- Controls: yes dilution water control only
- 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)): 5 µl/L

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: laboratory culture
- Feeding during test: yes
- Food type: unicellular green algae Selenastrum capricornutum and Tetrafin
- Frequency:daily

Test organisms were cultured at 22 °C with photoperiod 16L:8D
Test organisms were < 24 h old

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

not provided

Test temperature:

22 ± 1 °C for cultures

pH:

8.4 s.u.

Dissolved oxygen:

not given

Salinity:

NA

Nominal and measured concentrations:

Nominal concentrations control, 6.2, 12, 25, 50, 100 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 ml beakers with 40 ml test solution
- Renewal rate of test solution (frequency/flow rate): 3 times a week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: laboratory reconstituted, hardness not provided, but salt content as follows: 192 mg/L CaSO4, 192 mg/L NaHCO3, 120 mg/L MgSO4, 8 mg/L KCl
- Culture medium different from test medium: no
- Intervals of water quality measurement: not known

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D for cultures

VEHICLE CONTROL PERFORMED: uncertain

survival and reproduction assessed at renewal.

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

of parent animals

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.05 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

90-100% survival in all treatment groups
NOEC values are not specifically presented in the paper but inferred based on information and text provided. The chronic value of 71 µg/L is given in the text.

Reported statistics and error estimates:

none provided

Results for Reproduction of Daphnia magna Exposed to NP

NP (mg/L)	Live Offspring/female Early Phase	Live Offspring/female Early Phase
control	24 +/-13	20 +/-7
0.0062	18 +/-11	17 +/-13
0.012	27 +/-18	18 +/-13
0.025	26 +/-19	14 +/-7
0.05	14 +/-14	13 +/-10
0.10	6 +/-8*	16 +/-10

*significantly different from control (p=0.008)

Validity criteria fulfilled:

not specified

Conclusions:

The purpose of the study was to acertain the effects of nonylphenol on testosterone metabolic pathway. It was determined that nonylphenol does disrupt components of the testosterone metabolic pathway. It was also found that nonylphenol significantly decreases fecundity while having no effect on survival of parent organisms.

Executive summary:

The 21-day-chronic toxicity of nonylphenol to Daphnia manga was studied under static renewal conditions. Daphnids were exposed to control and nonylphenol at nominal concentrations of 0.0062, 0.012, 0.025, 0.05, 0.1 mg/L.  The 21-day NOEC based on reproduction was 0.05 mg/L nonylphenol and survival was >0.10 mg/L nonylphenol.  The sublethal effects included were number of offspring/surviving parent.  Production of offsprings in the treated groups indicated that nonylphenol had an effect on the reproduction at concentrations greater than 0.05 mg /L.  The most sensitive end point was reproduction.

 

This study is classified as an acceptable supporting study and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1^stinstar):< 24 h

Test Type (Flowthrough Static, Static Renewal):static renewal

 

NOEC (survival): >0.1 mg/L nonylphenol

NOEC (reproduction): 0.05 mg/L nonylphenol

 

Baldwin et al. (1997) was selected as a supporting study because it provides toxicity information on the sensitivity of the preferred test organism Daphnia magna to nonylphenol. Results of this study are consistent with test results of Brooke (1993), Brennen et al (2006), and the key study, Comber et al. (1993) for Daphnia magna.

Description of key information

 Hϋls (1992) reported a NOEC of 0.03 mg octylphenol/L for inhibition of D. magna (reproduction)
Marcial et al., (2003) reported a NOEC for survival of >0.01 mg octylphenol/L for Tigriopus japonicus. 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

0.03 mg/L

Marine water invertebrates

Marine water invertebrates

Dose descriptor:

NOEC

Effect concentration:

0.01 mg/L

Additional information

Reliable toxicity test results for long-term exposure of octylphenol to aquatic invertebrates included the key study by Hϋls (1992) using the freshwater preferred test species Daphnia magna, and the study by Marcial et al., (2003) using the marine copepod Tigriopus japonicus. Octylphenol exposure duration in each study was 21 days. Hϋls (1992) reported a NOEC of 0.03 mg octylphenol/L for inhibition of D. magna reproduction and Marcial et al., (2003) reported a NOEC for survival of >0.01 mg octylphenol/L for Tigriopus japonicus.