Quaternary ammonium compounds, C12-14-alkyl(hydroxyethyl)dimethyl, chlorides

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-07-20 to 1993-8-6

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was conducted according to OECD guideline 207 “Earthworm, Acute Toxicity Test". Study was well documented and GLP.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 207 (Earthworm, Acute Toxicity Tests)

Deviations:

no

Principles of method if other than guideline:

N/A

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

[see concordance under 1.1. of the CSR]

IUPAC Name:

[see concordance under 1.1. of the CSR]

Details on test material:

- Name of test material (as cited in study report): C12/14 KDB-Base (E.4408.01), C12/14 Alkyl dimethyl hydroxyethyl ammonium chloride
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): N/A
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): N/A
- Substance type: Active
- Physical state: white, transparent, liquid

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Vapour pressure: .NK
- Water solubility (under test conditions): good
- Henry's law constant: N/A
- log Pow: N/A
- pKa: N/A
- UV absorption: N/A
- pH dependance on stability: 7 +- 1.5


OTHER PROPERTIES (if relevant for this endpoint)
- Fate and behavior in the environment (where relevant to application patterns): N/A

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1000 mg of active ingredient per kg of dry soil.
- Sampling method: At the start of the test 100 ml samples from the control and the test substance stock solution containing 1000 mg a.i.l-1 were taken. The samples were taken in non-conditioned 200 ml glass bottles, after addition of 3 ml 37% v/v formaldehyde and 0.1 ml non-ionic preservative.
- Sample storage conditions before analysis: Samples were stored in a refrigerator until transfer to Procter and Gamble ETC.

Test substrate

Vehicle:

no

Details on preparation and application of test substrate:

- Method of application to filter paper (if used): N/A
- Method of mixing into soil (if used): To achieve a concentration of 1000 mg of active ingredient (a.i.) per kg of dry soil, 12500 mg of the test substance was accurately weighted and dissolved in 500 ml of demineralized water in a conditioned all glass container. A 50 ml aliquot of this solution together with 50 ml of demineralized water was added to 675 g of damp artificial soil and then thoroughly mixed. After mixing the soil was transferred into a test vessel.
- Method of application to soil surface (if used): N/A
- Controls: controls were prepared in a similar way by adding 100 ml of demineralized water to 675 g of damp artificial soil.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution): stock solution: N/A
- Evaporation of vehicle before use: N/A

Test organisms

Test organisms (species):

Eisenia fetida

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Common name: earthworm
- Source: worms had been bred from a stock originally supplied by NIAC-proefdierbedrijf, Millseweg 1, Beers (N-B), the Netherlands. They were grown in the lab in a horse manure- garden soil mix (1:1) at about 23 degree C.
- Age at test initiation (mean and range, SD): N/A
- Weight at test initiation (mean and range, SD): 0.3 - 0.6g


ACCLIMATION
- Acclimation period: 24 h prior to the test
- Acclimation conditions (same as test or not): same as test
- Health during acclimation (any mortality observed): N/A

Study design

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

yes

Total exposure duration:

14 d

Remarks:

Mortality was determined on days 7 and 14.

Post exposure observation period:

N/A

Test conditions

Test temperature:

21.4 degree C

pH:

pH at start (Control): 6.0
pH at start (highest test concentration) 6.1
pH at end (control): 6.2
pH at end (highest test concentration with survival): 6.2

Moisture:

Moisture content at start: 53% (based on dry constituents)
Moisture content at end: 54% (based on dry constituents)

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 1 L all-glass preserving jars with a glass lid.
- Amount of soil or substrate: 775 g (wet weight)
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: 0


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location:
- Pesticide use history at the collection site: N/A
- Collection procedures: N/A
- Sampling depth (cm): N/A
- Soil texture (if natural soil) N/A
- % sand:
- % silt:
- % clay:
- Soil taxonomic classification: N/A
- Soil classification system: N/A
- Composition (if artificial substrate): 1:2:7 (Sphagnum peat, Fine industrial sand, Kaolin clay)
See section any other information on materials and methods section below for more details.
- Organic carbon (%): N/A
- Maximum water holding capacity (in % dry weight): N/A
- CEC: N/A
- Pretreatment of soil: N/A
- Storage (condition, duration): N/A
- Stability and homogeneity of test material in the medium: N/A


OTHER TEST CONDITIONS
- Photoperiod: Continuous
- Light intensity: 760 lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality, weight, behavior and appearance, on days 7 & 14.


VEHICLE CONTROL PERFORMED: N/A


TEST CONCENTRATIONS
- Spacing factor for test concentrations: N/A
- Justification for using less concentration than requested by guideline: N/A
- Range finding study
- Test concentrations: 0, 10, 100, and 1000 mg a.i/kg
- Results used to determine the conditions for the definitive study: yes, the test was conducted as a separate range-finding test and a final test instead of one combined range-finding/limit test as was indicated in the protocol. Because no effects were expected from the range-finding test, the final test was conducted as a limit test, i.e. with one concentration of 1000 a.i. per kg dry soil.

Nominal and measured concentrations:

Nominal concentration of 1000 mg of active ingredient per kg of dry soil.
Nominal (control): 0 of active ingredient per kg of dry soil.
Measured concentrations: samples were taken, preserved, and sent to sponsor for analysis, but those results are not available in this study report.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

7 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 2

Duration:

14 d

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 3

Duration:

14 d

Dose descriptor:

LC100

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 4

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 5

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: behaviour and appearance

Effect concentrations 6

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

< 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: weight decrease

Details on results:

- Mortality at end of exposure period: none
- Total mass of adults at beginning of test: 21.2 g (control, 40 worms): 21.1 g (test treatment, 40 worms)
- Changes in body weight of live adults (% of initial weight) at end of exposure period: average percentage weight increase (s.d) for 1000 mg a.i.kg-1: -11.3 (2.5) as compared to the control -4.34 (0.874).
- No. of offspring produced: N/A
- No. of unhatched cocoons: N/A
- Morphological abnormalities: no
- Behavioral abnormalities: no
- Other biological observations: N/A

Results with reference substance (positive control):

N/A

Reported statistics and error estimates:

Two tailed Dunnett-test and p=95 and p=99 were calculated to show significant weight increase/decrease differences from that of the control worms.

Any other information on results incl. tables

Table 1. Summary of Earthworm Toxicity Test of KDB

Effect on	Parameter (14 days)	Nominal conc (mg/kg)
Mortality	NOEC	>1000
Mortality	LC50	>1000
Behavior & Appearance	NOEC	>1000
Weight decrease	NOEC	<1000

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

This study met the validity criteria of the OECD 207 Guideline

Conclusions:

The LC50 of C12-14 dimethyl hydroxyethyl ammonium chloride (KDB) to earthworms was > 1000 mg a.i./kg in a 14 day OECD 207 test (nominal concentration). Worms exposed to 1000 mg a.i..kg-1 showed a slightly lesser weight than the controls at the end of the test, resulting in a NOEC based on weight decrease of <1000 mg a.i./kg.

Executive summary:

The acute toxicity of C12-14 dimethyl hydroxyethyl ammonium chloride (KDB) to earthworms Eisenia fetida was determined in a 14 day test according to OECD Guideline no. 207. The study was carried out as a limit test according to the OECD principles of Good Laboratory Practice.

Worms were exposed for 14 days to the test substance mixed with an artificial soil at concentrations of 0 and 1000 mg of active ingredient per kg of dry soil (nominal test concentration). After 7 and 14 days, survival was determined, and mobility and appearance were visually assessed. The worms were also individually weighed at the start and the end of the experiment.

At 1000 mg a.i./kg of dry soil, no effects on mortality, behavior or appearance were observed after 14 days. As a result, the LC50 was >1000 mg/kg, and the NOEC based on each of these endpoints was >1000 mg/kg. A slight decrease in weight was observed in the populations exposed to 1000 mg (active ingredient)/kg after 14d. The NOEC, based on weight decrease, was <1000 mg/kg. The weight decrease was not evident in the range-finding test.

This toxicity test satisfies the guideline requirements for the OECD 207 Earthworm Acute Toxicity Test.