Quaternary ammonium compounds, C12-18-alkyltrimethyl, chlorides

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

From 08 August 1988 to 12 August 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no analytical monitoring

Justification for type of information:

Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

no

Details on sampling:

A clear stock solution of 15 mg/mL was prepared by diluting 0.15 g of the test substance with distilled water in a 10 mL flask. The appropriate volume of stock solution was then added to 15 L of dilution water in each test aquaria and mixed by stirring.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Chemical name of vehicle: Water
- Concentration of vehicle in test medium (stock solution): Stock suspension of 15 mg/L was prepared by diluting 0.15 g of the test substance with distilled water in a 10 mL flask.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Species: Salmo gairdneri (new name: Oncorhynchus mykiss)
- Source: Commercial supplier in California
- Length at study initiation (length definition, mean, range and SD): Mean = 33 mm; range = 27-40 mm
- Weight at study initiation (mean and range, SD): Mean = 0.31 g; range = 0.12- 0.58 g
- Feeding during test: No

ACCLIMATION
- Acclimation period: 14 d
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Dry commercial pelleted food, ad libitum
- Feeding frequency: Daily, except during the 48 h prior to testing
- Health during acclimation (any mortality observed): Yes (i.e., 1.1%)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

34 mg/L as CaCO3

Test temperature:

12-13 ⁰C

pH:

7.5

Dissolved oxygen:

67-72%

Nominal and measured concentrations:

Nominal: 0, 390, 670, 1,090, 1,820 and 3,030 µg/L (i.e., 0, 130, 223, 363, 607 and 1010 µg a.i./L)

Details on test conditions:

TEST SYSTEM
- Test vessel: 18.9 L glass aquaria which contained 15 L of the test solution
- Aeration: No
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): No data
- No. of vessels per control (replicates): No data
- Biomass loading rate: 0.21 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionized, reconstituted well water
- Alkalinity: 28 mg/L
- Conductivity: 150 µmhos/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: 24, 48, 72 and 96 h

OTHER TEST CONDITIONS
- Adjustment of pH: Not required
- Photoperiod: 16 h light and 8 h darkness
- Light intensity: 60 footcandles

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortalities were recorded at 0, 6, 24, 48, 72 and 96 h
TEST CONCENTRATIONS
- Spacing factor for test concentrations: <2
- Range finding study: Yes
- Test concentrations: 150 to 9,090 µg/L
- Results used to determine the conditions for the definitive study: After 96 h, 100% mortality at ≤1,520 µg/L.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2 350 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: i.e., equivalent to 783 µg a.i./L

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

3 030 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: i.e., equivalent to 1010 µg a.i./L

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1 820 µg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality, physical or behavioral abnormalities

Remarks on result:

other: i.e., equivalent to 607 µg a.i./L

Details on results:

After 96 h of exposure, 100% mortality was observed at the highest concentration (3,030 µg/L or 1010 µg a.i./L). No significant toxicant-related mortalities were observed at test concentrations ≤1820 µg/L or 607 µg a.i./L. The 96 h LC50 for rainbow trout exposed to the test substance was estimated by non-linear interpolation to be 2,350 µg/L, with a 95% confidence interval calculated by binomial probability of 1,820 - 3,030 µg/L. The NOAEC was established at 1,820 µg /L (or 607 µg a.i./L).





No significant toxicant-related mortalities were observed at test concentrations below 1010 µg a.i./l
No chemical analyses of testsubstance have been performed.
• TMAC is a quaternary ammonium compound which is not susceptible to hydrolyses or photodegradation.
• Stability of test solutions under test conditions has been shown in chronic daphnia tests with comparable quaternary compounds.
• There is no significant decrease observed over 3 days of testing.
• Quaternary ammonium compounds tend to adsorb to test organisms and glassware.
• Observed reduction of test concentration is maximal between 20 – 50 %


Conclusion:
The test substance was at least 50% available during the acute daphnia and fish tests.Further to the fish studies:
• The nominal effect data for fish are consistently about a factor 10 higher than for algae or daphnia.
• Repeating the acute fish tests will not improve the effect assessment for the aquatic compartment.
• Literature data on fish studies show comparable results
For ethical reasons we propose to calculate the effect data for fish assuming 50% availability as a worst-case.

Validity criteria were fulfilled, except for the concentration of the test substance during the test. See table 5
Test substance is moderately toxic according to EPA guidelines, but is very toxic according to European guidelines

Reported statistics and error estimates:

Probit analysis

Sublethal observations / clinical signs:

Deviation: The dissolved oxygen concentration were 75-77% of saturation (against 90% as per the protocol), however this deviation did not affect the results of the test.

Table 1:     Mortality data

Test­-Substance Concentration (nominal) [µg a.i./l]	Mortality
	Percentage    0 h          24 h         48 h         72 h         96 h
0		0	0	0^c	0^d
130		0	0	0	0
223		0	0	0^a	10
363		0^a	0^a	0^b	0^de
607		0	0^a	0^b	0^de
1010		100	100	100	100

^aOne of the surviving fish exhibited darkened pigmentation

^bTwo of the surviving fish exhibited darkened pigmentation

^cOne of the surviving fish exhibited darkened pigmentation and erratic swimming behavior

^dOne of the surviving fish exhibited a complete loss of equilibrium

^eSeveral of the surviving fish exhibited darkened pigmentation

 

Table 2: Oxygen concentrations

Test­-Substance Concentration (nominal) [µg a.i./l]	concentration (mg/l) and percentage    0 h          24 h         48 h         72 h         96 h
0	8.1 (77)	9.2 (85)	8.1 (77)	8.2 (78)	6.4 (64)
130	8.0 (76)	8.8 (81)	7.8 (74)	8.0 (76)	7.2 (68)
223	7.9 (75)	8.8 (81)	7.8 (74)	7.8 (74)	6.8 (64)
363	7.9 (75)	8.4 (78)	7.4 (70)	7.7 (73)	6.8 (64)
607	7.9 (75)	8.2 (76)	7.4 (70)	7.3 (69)	6.8 (64)
1010	7.9 (75)	8.2 (76)	-	-	-

 

Table 3: pH values

Test­-Substance Concentration (nominal) [µg a.i./l]	0 h          24 h         48 h         72 h         96 h
0	7.6	6.6	6.7	7.2	7.5
130	7.7	6.6	6.7	7.2	7.3
223	7.7	6.6	6.8	7.2	7.3
363	7.7	6.6	6.8	7.2	7.2
607	7.7	6.5	6.9	7.3	7.2
1010	7.7	6.6	-	-	-

 

Table 4: Effect data

	48 h [µg a.i./l]^1	95 % c.l.	96 h [µg a.i./l]^1	95 % c.l.
NOEC	-	-	607 (n)	-
LC50	783	607-1010	783	607-1010

¹indicate if effect data are based on nominal (n) or measured (m) concentrations

 

Table 5: Validity criteria for acute fish test according to OECD Guideline 203

	fulfilled	Not fullfilled
Mortality of control animals <10%	YES
Concentration of dissolved oxygen in all test vessels > 60% saturation	YES
Concentration of test substance³80% of initial concentration during test	*

*No analyses were performed, therefore it is not known if the test substance³80% of initial concentration during test

Validity criteria fulfilled:

yes

Remarks:

As per OECD guideline 203

Conclusions:

Based on the results, the 96 h LC50 and NOEC of the test substance were determined to be 2,350 and 1,820 µg a.i./L (i.e., equivalent to 783 and 607 µg a.i./L), respectively.

Executive summary:

A study was conducted to determine the acute toxicity study of the read across substance, Coco TMAC (33% active in water), to the Rainbow trout (Oncorhynchus mykiss) according to a method similar to OECD Guideline 203, in compliance with GLP. Ten Rainbow trouts per concentration were exposed to the read across substance at 0, 390, 670, 1,090, 1,820 and 3,030 µg/L (i.e., equivalent to 0, 130, 223, 363, 607 and 1010 µg a.i./L) for 96 h under static conditions. Mortalities were determined after 0, 24, 48 72 and 96 h. No analytical determination of the read across substance was performed. Throughout the test period no visible sign of undissolved material was observed in the exposure solutions. After 96 h of exposure, 100% mortality was observed at the highest concentration (3,030 µg/L or 1010 µg a.i./L). No significant toxicant-related mortalities were observed at test concentrations ≤1820 µg/L or 607 µg a.i./L. The 96 h LC50 for rainbow trout exposed to the read across substance was estimated by non-linear interpolation to be 2,350 µg/L, with a 95% confidence interval calculated by binomial probability of 1,820 - 3,030 µg/L. The NOAEC was established at 1,820 µg /L (or 607 µg a.i./L). Under the conditions of the study, the 96-h LC50 and NOEC were determined at 783 and 607 µg a.i./L respectively (Surprenant, 1988). Based on the results of the read across study, similar effect levels are considered for the test substance.

Description of key information

Due to the identified deficiency in the current study with the read across substance and as a conservative approach, the 96 h LC50 value of 0.14 mg a.i./L, in accordance with the biocides assessment report, has been considered further for hazard/risk assessment of the test substance. 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

140 µg/L

Additional information

study was conducted to determine the acute toxicity study of the read across substance, Coco TMAC (33% active in water), to the Rainbow trout (Oncorhynchus mykiss) according to a method similar to OECD Guideline 203, in compliance with GLP. Ten Rainbow trouts per concentration were exposed to the read across substance at 0, 390, 670, 1,090, 1,820 and 3,030 µg/L (i.e., equivalent to 0, 130, 223, 363, 607 and 1010 µg a.i./L) for 96 h under static conditions. Mortalities were determined after 0, 24, 48 72 and 96 h. No analytical determination of the read across substance was performed. Throughout the test period no visible sign of undissolved material was observed in the exposure solutions. After 96 h of exposure, 100% mortality was observed at the highest concentration (3,030 µg/L or 1010 µg a.i./L). No significant toxicant-related mortalities were observed at test concentrations ≤1820 µg/L or 607 µg a.i./L. The 96 h LC50 for rainbow trout exposed to the read across substance was estimated by non-linear interpolation to be 2,350 µg/L, with a 95% confidence interval calculated by binomial probability of 1,820 - 3,030 µg/L. The NOAEC was established at 1,820 µg /L (or 607 µg a.i./L). Under the conditions of the study, the 96-h LC50 and NOEC were determined at 783 and 607 µg a.i./L respectively (Surprenant, 1988). Based on the results of the read across study, similar effect levels are considered for the test substance.  

The biocide assessment report available from RMS Italy on Coco TMAC (ECHA assessment report, 2016), considered the above study to be supportive because the endpoints were based on nominal concentrations, which could underestimate the toxicity. However, several available reliable read across studies with DDAC and BKC, also indicated toxicity in the same range. As final endpoint for Coco TMAC, the RMS considered the read across data to DDAC with the lowest 96h LC50 = 0.19 mg a.s./L in Fathead minnow as the most sensitive species, which resulted in a 96h LC50 = 0.14 mg a.i./L for Coco TMAC, by applying correction for MW.  

Therefore, due to the identified deficiency and as a conservative approach, the 96 h LC50 value of 0.14 mg a.i./L, in accordance with the Coco TMAC biocides assessment report, has been considered further for hazard/risk assessment.