Docusate sodium

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 to 28 April 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP and guideline compliant study, deviations to the guideline were minor, e.g., the temperature ranged from 19 to 21 °C (guideline 20-24°C) and the fish had a length of 2 cm (guideline 3.0 +/- 0.5 cm). The stability of the test item under test conditions was determined in a seperate expermient.

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

but only minor: the temperature ranged from 19 to 21°C and the fish were smaller than stated in the guideline

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

The stock solution prepared with nominal 10 g Docusate Sodium/L was submitted to DOC analysis. The experimentally determined concentration was 8.884 g Docusate Sodium/L. Since the deviation to the nominal concentration was < 20 %, the nominal concentration of the stock solution was used for calculations. The stability of Docusate Sodium in test medium was determined in a separate experiement. In that experiment the test solutions were stable for at least 24 hours, i.e., the renewal interval of this fish test. The stability of the Docusate Sodium in the test solutions used to expose the fish was not determined.

Analysis of the stock solution and the stability check of the test solutions under similar conditions indicates that Docusate Sodium was (1) correctly dosed and (2) stable under test conditions, even if the test solutions used for the exposure were not analysed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution with 10 g Docusate Sodium/L (15.6 g SERWET WH 170/L) was prepared with deionized water.
- Controls: blank control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehicle used
- Concentration of vehicle in test medium (stock solution and final test solution): not applicable

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: West Aquarium, Bad Lauterberg, Germany
- Length at study initiation (length definition, mean, range and SD): 2.0 cm
- Weight at study initiation (mean and range, SD): about 0.25 g/fish




ACCLIMATION
- Acclimation period: at least 3 weeks, in the first week three treatments with malachit green were performed, thereafter 14 days quarantine
- Acclimation conditions (same as test or not): 200L or 300L culture vessels, 20 +/- 2°C water temperature, flow-through conditions
- Type and amount of food: TetraMin, 1 % of the fish weight per day
- Feeding frequency: daily
- Health during acclimation (any mortality observed): The fish showed nomal behaviour at the start of the test and had no obvious deseases. The mortality during the 7 days prior to start of the exposure was equal or less than 5%.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

about 13 °dH

Test temperature:

temperature range: 19 - 21°C, mean temperature: 20°C

pH:

7.7 to 8.2

Dissolved oxygen:

96 to 105% of the air saturation value

Nominal and measured concentrations:

Nonimal concentrations: 0 (control), 5, 10, 20, 40 and 80 mg Docusate Sodium/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Volume about 20 L, filled with 10 L test solution
- Aeration: continuous
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): NA
- Biomass loading rate: 0.25 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Drinking water provided by Gelsenwasser AG
- Culture medium different from test medium: no
- Intervals of water quality measurement: not clearly described


OTHER TEST CONDITIONS
- Photoperiod: 16:8 light-dark
- Light intensity: not provided


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
LC50 at 24, 48, 72, and 96 hours, LC0 and LC100 at 96-hours


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: NA

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

49 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: range: 40 - 80 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

49 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: range: 40 - 80 mg/L

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

49 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: range: 40 - 80 mg/L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

49 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: range: 40 - 80 mg/L

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

20 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

80 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: none

Reported statistics and error estimates:

graphical evaluation (concentration vs. % inhibition)

Sublethal observations / clinical signs:

Table 1  Mean temperatures and temperature ranges measured in the test vessels

 

Concentration mg Docusate Sodium/L	Temperature °C
	Mean	Min	Max
0	20	20	21
5	20	19	20
10	20	19	20
20	20	19	20
40	20	19	20
80	20	19	20

 

Table 2   Dissolved oxygen air saturation values and pH values measured in the test vessels at different time intervals

 

Concentration mg Docusate Sodium/L	Dissolved Oxygen (%) and pH
	0-hour test solution				24-hour test solution		48-hour test solution		72-hour test solution
	0-h new		after 24 h
	O2	pH	O2	pH	O2	pH	O2	pH	O2	pH
0	97	7.7	105	8.2	96	7.9	101	8	100	7.9
5	97	7.7	104	8.2	97	7.9	100	8	98	7.9
10	96	7.7	104	8.2	96	7.9	101	8	98	7.9
20	97	7.7	104	8.2	97	7.9	101	7.9	98	7.9
40	96	7.7	102	8.2	97	7.9	101	7.9	98	7.9
80	96	7.7	104	8.2	-*	-*	-*	-*	-*	-*

 

* discontinued due to 100% mortality

 

Table 3 Number of dead and surving fish and % mortality at the individual observation intervals

 

Concentration mg Docusate Sodium/L	Time interval
	24			48			72			96
	Number		% dead	Number		% dead	Number		% dead	Number		% dead
	alive	dead		alive	dead		alive	dead		alive	dead
0	10	0	0	10	0	0	10	0	0	10	0	0
5	10	0	0	10	0	0	10	0	0	10	0	0
10	10	0	0	10	0	0	10	0	0	10	0	0
20	10	0	0	10	0	0	10	0	0	10	0	0
40	7	3	30	7	3	30	7	3	30	7	3	30
80	0	10	100	0	10	100	0	10	100	0	10	100

 

Table 4   LC0, LC50 and LC100 values for Docusate Sodium

Time interval	For Docusate Sodium (mg/L)
	LC50	Range
		lower	upper
24 h	49	40	80
48 h	49	40	80
72 h	49	40	80
96 h	49	40	80
96-h LC0	20 mg/L
96-h LC100	80 mg/L

Validity criteria fulfilled:

yes

Conclusions:

96-h LC50=49mg Docusate Sodium/L

Executive summary:

The acute toxicity of SERWET WH 170 - which contains 64.2% Docusate Sodium - to zebra fish, Brachydanio rerio (new Danio rerio) was determined in a semi-static test with daily renewal of the test solutions. The test was performed according to EG 92/69 C.1. Deviations to the guideline were minor, e.g., the temperature ranged from 19 to 21 °C (guideline 20-24°C) and the fish had a length of 2 cm (guideline 3.0 +/- 0.5 cm). The test was performed with a control and nominal 5, 10, 20, 40 and 80 mg Docusate Sodium/L. The stock solution was prepared with nominal 10 g Docusate Sodium/L and was subsequently submitted to DOC analysis. The experimentally determined concentration was 8.884 g Docusate Sodium/L. Since the deviation to the nominal concentration was < 20 %, the nominal concentration of the stock solution was used for calculations on dosing. The stability of the Docusate Sodium in test medium was determined in a separate side experiment similar to the study with Brachydanio rerio. In that experiment without fish the test solutions were stable for at least 24 hours, i.e., the renewal interval of this test. During the study, the temperature of the test solutions ranged from 19 to 21°C with an average temperature of 20°C. After 96 hours of exposure, 0, 0, 0, 0, 3, and 10 out of 10 fish had died in the control, 5, 10, 20, 40 and 80 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 49 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

The results of this study are considered to be relevant for the risk assessment.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

November 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

GLP and test guideline compliant study, good documentation, no dose verification performed

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The definitive test concentrations were obtained by transfering appropriate weights of test item directly to the test chambers.
- Controls: blank controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): NA
- 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): All solutions from 32 to 100 mg/L had a small amount of undissolved compound of undissolved compound at 0-hour, which increased slightly with an increase in concentration. After 24-hours, a small amount of undissolved compound was still present at 56, 75 and 100 g/L. After 48- and 72-hours, the 42 mg was slightly cloudy, no solution observations were recorded at 96-hours.

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: bluegill sunfish
- Source: Osage Catfisheries, Osage Beach
- Length at study initiation (length definition, mean, range and SD): 22 (+/- 3.7 ) mm
- Weight at study initiation (mean and range, SD): 0.27 (+/- 0.16) g/fish
- Feeding during test: the fish were not fed durung the exposure



ACCLIMATION
- Acclimation period: at least 14 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: standrad commercial fish food
- Feeding frequency: daily, discontinued 48 to 96 hours prior to testing


QUARANTINE (wild caught)
- Duration: at least 14 days

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

225- 275 ppm

Test temperature:

22°C

pH:

7.1 - 7.6

Dissolved oxygen:

5.9 - 8.4 mg/L

Nominal and measured concentrations:

nominal concentrations: 0 (control), 32, 42, 56, 75 and 100 mg CT-326-87/L, corresponding to 0 (control), 31.0, 40.7, 54.3, 72.8, 97.0 mg Docusate Sodium/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 gallon glass vessels containing 15 L test solution
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): NA
- Biomass loading rate: 0.18 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: soft reconstituted water
Following amounts were added to deionized water:
NaHCO3: 48 mg/L
CaSO4*2H2O: 30 mg/L
MgSO4: 30 mg/L
KCl: 2 mg/L
- Metals: measured, levels below levels of concern
- Pesticides:measured, levels below levels of concern
- Alkalinity: 325 - 375 mg/L as CaCO3
- Conductivity: 700 µmohs/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: measured at 24, 48 and 96 hours


OTHER TEST CONDITIONS
- Adjustment of pH: no



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality and behavioral changes

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3
- Justification for using less concentrations than requested by guideline: NA
- Range finding study: two range finding studies were performed:
- Test concentrations:
first range finder: 1, 10, 100 mg CT-326-87/L
second range finder: 32 mg CT-326-87/L
- Results used to determine the conditions for the definitive study:
first range finder: 1, 10, 100 mg CT-326-87/L with 0/5, 0/5, 5/5 dead fish after 96 hours;
second range finder: 32 mg CT-326-87/L with 0/5 dead fish after 96 hours

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

36.9 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 31.0 - 40.7 mg Docusate Sodium/L

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

38 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 32 - 42 mg test material/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

36.9 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 31.0 - 40.7 mg Docusate Sodium/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

38 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 32 - 42 mg test material/L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

35.9 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 31.0 - 40.7 mg Docusate Sodium/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

37 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 32 - 42 mg test material/L

Details on results:

- Behavioural abnormalities: none reported
- Observations on body length and weight: NA
- Other biological observations: no behavoral changes at 32 mg CT-326-87/L
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: All solutions from 32 to 100 mg/L had a small amount of undissolved compound of undissolved compound at 0-hour, which increased slightly with an increase in concentration. After 24-hours, a small amount of undissolved compound was still present at 56, 75 and 100 g/L. After 48- and 72-hours, the 42 mg was slightly cloudy, No solution observations were recorded at 96-hours.
- Effect concentrations exceeding solubility of substance in test medium: The water solubility of Docusate Sodium was given as 1.5 g/100 mL. Therefore, the used concentrations should be below the water solubility limit and hence, the precipitated compound is unlikely to be Docusate Sodium but less soluble impurity.

Reported statistics and error estimates:

The LC50 values for CT-326-87 were determined using the binomial method.

Sublethal observations / clinical signs:

Table 1 Nominal concentration of CT-326-87 and corresponding concentrations of Docusate Sodium in the test solutions (calculated for 97% purity)

 

Nominal concentration [mg CT-326-87/L]	Nominal concentration [mg Docusate Sodium/L]
0 (Control)	0 (Control)
32	31.0
42	40.7
56	54.3
75	72.8
100	97.0

 

 

Table 2   Temperature, dissolved oxygen and pH values measured in the test vessels at different time intervals

 

Nominal concentration [mg CT-326-87 /L]	24 hours			48 hours			96 hours
	Temp [°C]	DO [mg/L]	pH	Temp [°C]	DO [mg/L]	pH	Temp [°C]	DO [mg/L]	pH
0 (Control)	22	8.3	7.6	22	6.5	7.2	22	6.2	7.2
32	22	8.3	7.6	22	6.3	7.1	22	5.9	7.1
42	-	-	-	22	6.1	7.1	22	6.1	7.2
56	22	8.3	7.5	Discontinued
75	-	-	-
100	22	8.4	7.6

 

 

Table 3 Percent mortality at the individual observation intervals

 

Nominal concentration [mg CT-326-87 /L]	Cumulative Percent Mortality
	24 hours	48 hours	96 hours
0 (Control)	0	0	0
32	0	0	0
42	90	90	100
56	100	100	100
75	100	100	100
100	100	100	100

 

 

Table 4   LC50 values for CT-326-87 and Docusate Sodium

Time interval	For CT-326-87 (mg/L)
	LC50	95% C.I.
		Lower	upper
24 h	38	32	42
48 h	38	32	42
96 h	37	32	42

 

Time interval	For Docusate Sodium (mg/L)
	LC50	95% C.I.
		Lower	upper
24 h	36.9	31.0	40.7
48 h	36.9	31.0	40.7
96 h	35.9	31.0	40.7

 

The LC50 values for CT-326-87 were determined using the binomial method, the LC50 values for Docusate Sodium were calculated from theCT-326-87LC50 values and the purity of 97%. 

Validity criteria fulfilled:

yes

Conclusions:

96-hour LC50 (Lepomis macrochirus)=35.9 mg Docusate Sodium/L

Executive summary:

The acute toxicity of CT-326 -87 - which contains at least 97% Docusate Sodium - to bluegill sunfish, Lepomis macrochirus, was determined in a static test. The test was performed according to OECD 203. Deviations to the guideline were not reported. No dose-verification analysis was performed. The test was performed with a control and nominal 32, 42, 56, 75 and 100 mg CT-326 -87/L corresponding to 31.0, 40.7, 54.3, 72.8, 97.0 mg Docusate Sodium/L. During the study, the temperature of the test solutions was 22°C. After 96 hours of exposure, 0, 0, 10, 10, 10 and 10 out of 10 fish had died in the control, and 31.0, 40.7, 54.3, 72.8, 97.0 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 35.9 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

The results of this study are considered to be reliable with restrictions for the risk assessment.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

27 - 31 January 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

GLP compliant study, no dose verification analysis

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1400 (Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: fathead minnow
- Strain: in-house culturing program
- Source: laboratory breed
- Age at study initiation (mean and range, SD): 72 - 75 days
- Length at study initiation (length definition, mean, range and SD): mean length 27 mm
- Weight at study initiation (mean and range, SD): 0.136g/fish (wet weight)
- Method of breeding: All of the fish were maintained in holding tanks at 22 to 24°C with 16-hour light and 8-hour dark photoperiod. These fish were fed brine shrimp nauplii (Artemia sp.) and Tetramin fish food twice a day except for some occational fasting on weekends. The test fish received Tetramin about two weeks after hatching. Test fish were removed from feed 24 hours prior to start of the study.
- Feeding during test: none



ACCLIMATION
- Acclimation period: none
- Acclimation conditions (same as test or not): Similar to test conditions
- Type and amount of food: The fish were fed brine shrimp nauplii (Artemia sp.) and Tetramin fish food
- Feeding frequency: The fish were fed twice a day except for some occational fasting on weekends
- Health during acclimation (any mortality observed): The study plan states that fish are judged healthy if the mortality is <5% within one week prior to start of the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

60 mg/L as CaCO3

Test temperature:

See Table 1

pH:

See Table 1

Dissolved oxygen:

See Table 1

Nominal and measured concentrations:

The test concentrations were 0 (control), 0.625, 1.25, 2.5, 5, 10, 20 and 40 mg TRITON GR-7M Surfactant corresponding to 0 (control),0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 mg Docusate Sodium/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): 2000 mL beakers
- Material, size, headspace, fill volume: fill volume: 1500 mL
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): NA
- Biomass loading rate: 0.91 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated Charleston municipal water
- Intervals of water quality measurement: at start and every 24 hours


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16:8 light:dark



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and sublethal effects every 24 hours after start of the exposure

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: NA
- Range finding study: Not performed, test concentrations were selected based on information from a Daphnia range finding study and experience with similar substances

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

17.3 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Docusate Sodium

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 16.6 - 18.6 mg/L

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 26 - 29 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

17.3 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Docusate Sodium

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 16.6 - 18.6 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 26 - 29 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

17.3 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Remarks:

Docusate Sodium

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 16.6 - 18.6 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I. 26 - 29 mg/L

Details on results:

- Behavioural abnormalities: none
- Observations on body length and weight: not performed
- Other biological observations: NA
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none

Reported statistics and error estimates:

The LC50 was estimated using the Spearman Karber estimator on the percent mortalities for each concentration and the control.

Sublethal observations / clinical signs:

Table 1 Dissolved oxygen concentration, pH and temperature measured during the exposure

Concentration mg TRITON GR-7M Surfactant/L	Repl.	0-hour			24-hour			48-hour			72-hour			96-hour
		DO	pH	Temp	DO	pH	Temp	DO	pH	Temp	DO	pH	Temp	DO	pH	Temp
		mg/L		°C	mg/L		°C	mg/L		°C	mg/L		°C	mg/L		°C
Control	A	8.1	7.6	22.4	6.8	6.6	22.3	5.2	7.0	21.4	5.4	6.2	22.2	6.0	6.7	21.6
	B	8.2	7.5	22.2	4.6	6.6	22.3	5.3	7.0	21.2	5.9	6.4	22.1	6.3	6.7	21.6
0.625	A	8.1	7.5	22.1	4.7	6.6	22.3	5.3	6.9	21.3	6.0	6.3	22.0	6.4	6.6	21.5
	B	8.2	7.5	22.1	4.7	6.6	22.3	5.2	7.0	21.3	5.9	6.4	22.0	6.4	6.7	21.5
1.25	A	8.1	7.5	22.1	4.8	6.6	22.3	5.3	6.8	21.3	6.1	6.5	22.0	6.3	6.6	21.5
	B	8.2	7.5	22.1	5.0	6.5	22.4	5.8	6.8	21.4	5.8	6.6	22.3	5.9	6.6	21.5
2.5	A	8.1	7.5	22.1	4.8	6.5	22.3	5.6	6.8	21.2	5.8	6.7	22.1	7.0	6.6	21.5
	B	8.2	7.4	22.1	4.9	6.5	22.3	5.7	6.8	21.2	5.8	6.7	22.0	6.4	6.6	21.5
5	A	8.2	7.4	22.1	5.1	6.5	22.3	5.7	6.8	21.2	5.6	6.7	22.0	6.6	6.7	21.5
	B	7.8	7.4	22.1	5.1	6.5	22.3	5.8	6.8	21.2	5.5	6.7	21.9	5.7	6.7	21.5
10	A	7.8	7.4	22.1	4.6	6.5	22.3	5.7	6.8	21.2	5.8	6.8	22.0	5.9	6.6	21.5
	B	8.2	7.4	22.1	4.1	6.5	22.5	5.7	6.7	21.3	5.4	6.8	22.3	5.9	6.7	21.5
20	A	7.7	7.4	22.2	5.4	6.5	22.1	5.0	6.7	21.2	5.3	6.7	22.1	5.7	6.6	21.5
	B	7.7	7.4	22.2	5.2	6.5	22.2	4.5	6.7	21.2	5.1	6.7	21.9	5.9	6.6	21.5
40	A	7.6	7.4	22.5	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead
	B	7.6	7.3	22.2	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead	Dead

Table 2 Mortaility of Pimephales promeals during the exposure to TRITON GR-7M Surfactant (each replicate was started with 10 fish) and corresponding LC50 values

Concentration mg TRITON GR-7M Surfactant/L	Repl.	24-hour		48-hour		72-hour		96-hour
		Resp	Dead	Resp	Dead	Resp	Dead	Resp	Dead
Control	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
0.625	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
1.25	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
2.5	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
5	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
10	A	N	-	N	-	N	-	N	-
	B	N	-	N	-	N	-	N	-
20	A	N	1	N	1	N	1	N	1
	B	N	-	N	-	N	-	N	-
40	A	-	10	-	10	-	10	-	10
	B	-	10	-	10	-	10	-	10
LC50 (mg TRITON GR-7M Surfactant /L)		27		27		NA		27
Lower/upper (mg TRITON GR-7M Surfactant /L)		26 / 29		26 / 29		NA / NA		26 / 29
Observed no effect level (mg TRITON GR-7M Surfactant /L)		10		10		NA		10
N: Nothing unusual observed

Table 3 Observed no effect levels and LC50 values based on Docusate Sodium

Time interval [h]

24

48

72

96

LC50 (mg Docusate Sodium /L)		17.3		17.3		NA		17.3
Lower/upper (mg Docusate Sodium /L)		16.6 / 18.6		16.6 / 18.6		NA / NA		16.6 / 18.6
Observed no effect level (mg Docusate Sodium /L)		6.4		6.4		6.4		6.4

Validity criteria fulfilled:

yes

Executive summary:

The acute toxicity of TRITON GR-7M Surfactant - which contains 64% Docusate Sodium - to fathead minnow, Pimephales promelas, was determined in a static 96 hour test. The test was performed according to U.S. EPA-TSCA, 40 CFR Part 797.1400. The test concentrations were 0 (control), 0.625, 1.25, 2.5, 5, 10, 20 and 40 mg TRITON GR-7M Surfactant/L corresponding to 0 (control),0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 mg Docusate Sodium/L. Dose verification was not performed and therefore the concentrations were based on nominal concentrations. The test was set up with two replicates per test group. Each replicate received 10 fish, i.e., the 20 fish were exposed per test group. The temperature ranged from 21 to 23°C, the dissolved oxygen concentration from 4.1 to 8.2 mg/L, the pH from 6.2 to 7.6. After 96 hours of exposure, 0, 0, 0, 0, 0, 0, 1 and 20 out of 20 fish had died in the control, 0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 27 mg TRITION GR-7M Surfactant/L corresponding to 17.3 mg Docusate Sodium/L.

The results of this study are considered to be reliable with restrictions for the risk assessment.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

7 - 24 November 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Guideline compliant GLP study. Concentrations reported as nominal concentrations since no dose verification analysis was performed.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout
- Source: Aquatic Research Organisms, Hampton, NH
- Age at study initiation (mean and range, SD): 70 days
- Length at study initiation (length definition, mean, range and SD): smaller or equal 50 mm, uniform size

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

90 mg/L as CaCO3

Test temperature:

15°C

pH:

see Table 2

Dissolved oxygen:

see Table 2

Nominal and measured concentrations:

Nominal: 0 (control), 6.25, 12.5, 25, 50 and 100 mg CT 444 90 C/L, corresponding to 6.06, 12.1, 24.3, 48.5 and 97 mg Docusate Sodium/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 4L polypropylene vessels
- Material, size, headspace, fill volume: fill volume 3000 mL
- Aeration: Oil free air was provided with <= 100 bubbles per minute to maintain >= 60 % saturation
- No. of organisms per vessel: Minimum 5 fish per replicate
- No. of vessels per concentration (replicates): 2 replicates
- No. of vessels per control (replicates):2 replicates
- No. of vessels per vehicle control (replicates): NA
- Biomass loading rate: <= 1 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: US EPA moderately hard reconsituted
- Alkalinity: 80 mg/L as CaCO3
- Conductivity:see table 2
- Intervals of water quality measurement: temperature, dissolved oxygen, pH, conductivity were measured initally and daily thereafter. Alkalinity and hardness of dilution water were measured at test initiation.


OTHER TEST CONDITIONS
- Photoperiod: 16 hr/8 hr light/dark cycle
- Light intensity: 50 to 100 foot-candles (lab ambient)


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality, behavior (lethargy, erratic swimming), and physiology (discoloration), initially, and daily thereafter

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: NA
- Range finding study: 5 fish per replicate exposed for 48 hours.
- Test concentrations: 0, 1, 10, 100 and 1000 mg CT 444 90 C/L
- Results used to determine the conditions for the definitive study: Mortality after 48 hours: 0/5, 0/5, 0/5, 5/5, 5/5 dead fish at 0, 1, 10, 100 and 1000 mg CT 444 90 C/L.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

34.9 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

36 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

27.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

28 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

27.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

28 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

27.2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

28 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Observations on body length and weight: NA
- Other biological observations: all of the fish exposed to 50 and 100 mg CT 444 90 C/L were dead within one hour. These fish demonstrated extreme stress immediately upon exposure.
- Mortality of control: none

Sublethal observations / clinical signs:

Table 1 Results from the 48 h range finding experiment

Nominal concentration [mg CT 444 90 C/L]	Dead fish vs. total exposed
0 (control)	0/5
1	0/5
10	0/5
100	5/5
1000	5/5

 

 

Table 2   Dissolved oxygen and pH values measured in the test vessels at different time intervals

 

Nominal concentration [mg CT 444 90 C /L]	24 hours			48 hours			72 hours			96 hours
	DO	pH	Cond.	DO	pH	Cond.	DO	pH	Cond.	DO	pH	Cond.
0 (Control)	9.8	7.0	210	9.4	7.5	220	9.8	7.3	220	9.6	7.4	220
6.25	9.8	6.8	210	8.0	7.1	250	8.6	6.5	220	8.8	7.0	220
12.5	9.8	7.0	220	8.0	7.4	230	8.6	6.7	220	8.8	6.9	220
25	9.8	7.1	220	8.0	7.3	220	8.6	6.7	220	8.8	6.9	220
50	9.8	7.2	220	Discontinued
100	9.8	7.3	210

 

 

Table 3 Number of dead fish and % mortality at the individual observation intervals

Nominal concentration [mg CT 444 90 C /L]	Number exposed	Cumulative Mortality				% mortality at 96 hours
		24 hours	48 hours	72 hours	96 hours
0 (Control)	10	0	0	0	0	0
6.25	10	0	0	0	0	0
12.5	10	0	0	0	0	0
25	10	0	2	2	2	20
50	10	10	10	10	10	100
100	10	10	10	10	10	100

 

 

Table 4   LC50 values for CT 444 90 C

Time interval	For CT 444 90 C (mg/L)
	LC50	Range
		lower	upper
24 h	36	NA	NA
48 h	28	NA	NA
72 h	28	NA	NA
96 h	28	NA	NA

Since the LC50 was determined using a graphical method, the 95% C.I. could not be provided.

Validity criteria fulfilled:

yes

Conclusions:

96-hour LC50= 27.2 mg Docusate Sodium/L

Executive summary:

The acute toxicity of CT 444 90 C - which contains at least 97% Docusate Sodium - to rainbow trout, Oncorhynchus mykiss, was determined in a static test. The test was performed according to OECD 203 without deviations to the guideline. The test was performed with a control and nominal 6.25, 12.5, 25, 50 and 100 mg CT 444 90 C/L corresponding to 6.06, 12.1, 24.3, 48.5 and 97 mg Docusate Sodium/L. For each test group, 2 replicates with 5 fish were set up. Dose verification analysis was not performed. During the study, the temperature of the test solutions ranged within 15 +/- 2°C. After 96 hours of exposure, 0, 0, 0, 2, 10 and 10 out of 10 fish had died in the control, 6.06, 12.1, 24.3, 48.5 and 97 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 27.2 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

The results of this study are considered to be reliable with restrictions for the risk assessment.

Description of key information

LC50 for freshwater fish (zebra fish) = 49 mg Docusate Sodium/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

49 mg/L

Additional information

For the data endpoint “Short-term toxicity to fish” in total 7 studies on freshwater fish are available: one Klimisch 1 key study, three supporting studies and three disregarded studies with Klimisch 3 or Klimisch 4 scores.

 

The study from Elementis (Scholz 1995) was classified as Klimisch 1 key study. The study was conducted according to the appropriate guidelines, without major deviations. In this study the acute toxicity of a formulation containing 64.2% Docusate Sodium to zebra fish, Brachydanio rerio (new Danio rerio) was determined in a semi-static test with daily renewal of the test solutions. The test was performed according to EG 92/69 C.1. Deviations to the guideline were minor, e.g., the temperature ranged from 19 to 21 °C (guideline 20-24°C) and the fish had a length of 2 cm (guideline 3.0 +/- 0.5 cm). The test was performed with a control and nominal 5, 10, 20, 40 and 80 mg Docusate Sodium/L. The stock solution was prepared with nominal 10 g Docusate Sodium/L and was subsequently submitted to DOC analysis. The experimentally determined concentration was 8.884 g Docusate Sodium/L. Since the deviation to the nominal concentration was < 20 %, the nominal concentration of the stock solution was used for calculations on dosing. The stability of the Docusate Sodium in test medium was determined in an appropriate separate side experiment under conditions similar to the study itself. In that experiment the test solutions were stable for at least 24 hours, i.e., the renewal interval of this test. During the study, the temperature of the test solutions ranged from 19 to 21°C with an average temperature of 20°C. After 96 hours of exposure, 0, 0, 0, 0, 3, and 10 out of 10 fish had died in the 0 (control), 5, 10, 20, 40 and 80 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 49 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

 

Since this study represents the technically and scientifically best study, it is considered reliable and relevant for the risk assessment of Docusate Sodium.

 

Beside the key study, three supporting Klimisch 2 and three disregarded Klimisch 3 or 4 studies are available for this endpoint. These studies are considered to be just supportive or disregarded since no dose verification was performed or major deviations to the guidelines were observed, i.e., the size of the fish was higher than recommended in the guideline and the test duration was only 48 hours instead of 96 hours as outlined in the guideline.

 

The Cytec study from Bowman (1987) was classified as Klimisch 2 key study. In this study the acute toxicity of test substance - which contains at least 97% Docusate Sodium - to bluegill sunfish, Lepomis macrochirus, was determined in a static test. The test was performed according to OECD 203. Deviations to the guideline were not reported. No dose-verification analysis was performed. The test was performed with a control and nominal 32, 42, 56, 75 and 100 mg CT-326 -87/L corresponding to 31.0, 40.7, 54.3, 72.8, 97.0 mg Docusate Sodium/L. During the study, the temperature of the test solutions was 22°C. After 96 hours of exposure, 0, 0, 10, 10, 10 and 10 out of 10 fish had died in the control, and 31.0, 40.7, 54.3, 72.8, 97.0 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 35.9 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

 

The Dow study from Raghuram (1992) was classified as Klimisch 2 key study. In this study the acute toxicity of a formulated product- which contains 64% Docusate Sodium - to fathead minnow, Pimephales promelas, was determined in a static 96 hour test. The test was performed according to U.S. EPA-TSCA, 40 CFR Part 797.1400. The test concentrations were 0 (control), 0.625, 1.25, 2.5, 5, 10, 20 and 40 mg formulated product/L corresponding to 0 (control),0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 mg Docusate Sodium/L. Dose verification was not performed and therefore the concentrations were based on nominal concentrations. The test was set up with two replicates per test group. Each replicate received 10 fish, i.e., the 20 fish were exposed per test group. The temperature ranged from 21 to 23°C, the dissolved oxygen concentration from 4.1 to 8.2 mg/L, the pH from 6.2 to 7.6. After 96 hours of exposure, 0, 0, 0, 0, 0, 0, 1 and 20 out of 20 fish had died in the control,0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 17.3 mg Docusate Sodium/L.

 

The Cytec study from Drozdowski (1990) was classified as Klimisch 2 key study. In this study the acute toxicity of the test substance - which contains at least 97% Docusate Sodium - to rainbow trout, Oncorhynchus mykiss, was determined in a static test. The test was performed according to OECD 203 without deviations to the guideline. The test was performed with a control and nominal 6.25, 12.5, 25, 50 and 100 mg test substance/L corresponding to 6.06, 12.1, 24.3, 48.5 and 97 mg Docusate Sodium/L. For each test group, 2 replicates with 5 fish were set up. Dose verification analysis was not performed. During the study, the temperature of the test solutions ranged within 15 +/- 2°C. After 96 hours of exposure, 0, 0, 0, 2, 10 and 10 out of 10 fish had died in the control, 6.06, 12.1, 24.3, 48.5 and 97 mg Docusate Sodium/L treatment groups. The 96 h LC50 was determined to be 27.2 mg Docusate Sodium/L. The concentrations were based on nominal concentrations.

 

In addition to the Klimisch 1 and Klimisch 2 studies, there are two Klimisch 3 and one Klimisch 4 studies for short-term toxicity to fish available. Due to the poor reliability of these studies, they are not discussed here.

In contrast to the above key study from Elementis (Scholz, 1995), no dose verification was performed in the supporting studies (Klimisch 2) resulting in lower certainty for correct dosage when compared to the key study. The individual LC50 values of those studies were 35.9, 17.3 and 27,2 mg Docusate Sodium/L and therefore lower than those of the key study. It may be argued that a more conservative LC50 from one of these Klimisch 2 supporting studies could have been selected. However, as it will be outlined in the endpoint summary for Aquatic Toxicology (chapter 6.1), the EC50 from daphnids is below the lowest LC50 from fish - regardless if the result was obtained from the key-study or one of the supporting studies. Hence, the decision of the key study for the assessment of the toxicity on fish has no impact on the PNEC aquatic (freshwater) derivation. For clarity, the study with the best test design was selected as key study.

 

Therefore, the acute LC50 of 49 mg Docusate Sodium /L from the key study will be used for the further risk assessment of Docusate Sodium.