Docosyltrimethylammonium methyl sulphate

Toxicity to terrestrial arthropods

Administrative data

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

key study

Study period:

2008-04-18 to 2009-07-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According ISO 11267 Guideline and GLP. Justification for read-across see chemical safety report chapter 1.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

ISO 11267 (Inhibition of Reproduction of Collembola by Soil Pollutants)

Version / remarks:

and OECD Draft (2008)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Application method:

soil

Test material

Test material information

Constituent 1

Reference substance name:

Quaternary ammonium compounds, C20-22-alkyltrimethyl, chlorides

EC Number:

271-756-9

EC Name:

Quaternary ammonium compounds, C20-22-alkyltrimethyl, chlorides

Cas Number:

68607-24-9

IUPAC Name:

68607-24-9

Constituent 2

Reference substance name:

C20/22 ATQ

IUPAC Name:

C20/22 ATQ

Sampling and analysis

Analytical monitoring:

no

Test substrate

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil: Before application, the soil was moistened with deionised water to a water content of 60 % of thr maximum water capacity of the soil. The respective test item concentrations were weighed out and dissolved in 2.5 mL methanol each. The solution was mixed thoroughly with 10 g quartz sand in a glass vessel. After complete evaporation, the treated quartz sand was given to the artificial soil. After application test medium was mixed thoroughly with a blender for at least three minutes and 30 g wet weight was filled into each replicate.
- Controls: Artificial soil moistened with deionised water without test of reference item.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): organic solvent, methanol
- Concentration of vehicle in test medium (stock solution and final test solution): 2.5 mL per treatment
- Evaporation of vehicle before use: Yes, complete evaporation
- Volume of test solution applied: Mixture 10 g quartz sand and respective amount of test substance

Test organisms

Test organisms (species):

Folsomia candida

Animal group:

Collembola (soil-dwelling springtail)

Details on test organisms:

TEST ORGANISM
- Common name: Springtail, Folsomia candida WILLEM
- Source: Own breeding
- Age at test initiation (mean and range, SD): 12 days old


ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Feeding: The springtails were fed with granulated dry yeast
- Health during acclimation (any mortality observed): No mortality observed

Study design

Study type:

laboratory study

Limit test:

no

Total exposure duration:

28 d

Post exposure observation period:

None

Test conditions

Test temperature:

18 - 22 °C

pH (if soil or dung study):

pH-Values of the Soil on Day 0 and Day 28


Test item [mg/kg DW]
pH
Day 0 Day 28
Control 6.38 6.14
Solvent Control 6.43 6.32
62.5 6.44 6.32
125 6.44 6.29
250 6.44 6.32
500 6.44 6.28
1000 6.43 6.25

Humidity:

Soil Moisture [%] on Day 0, 14 and 28


Test item [mg/kg DW]
Soil moisture [%]1)
Day 0 Day 14* Day 28
Control 40.49 39.28 37.98
Solvent Control 40.18 39.19 42.08
62.5 39.32 37.11 34.57
125 39.05 36.33 32.86
250 38.93 37.67 38.99
500 38.57 36.72 40.53
1000 39.03 36.62 33.19
1 ) = Related to dry weight (DW) of the soil; maximum water holding capacity (WHC) = 71.5 g / 100 g DW
* ) = Due to difference > 2 %, additional moistening of the replicates with each 1 mL demin. water on day 16

Photoperiod and lighting:

16 h : 8 h (light : dark)
423 - 581 lx

Details on test conditions:

TEST SYSTEM
- Test container (material, size): Glass beakers, volume 100 mL (inner diameter: 4.3 cm), covered with Parafilm
- Amount of soil or substrate: 30 g wet weight each
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 5
- No. of replicates per control: 8
- No. of replicates per vehicle control: 8


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)

- Composition (if artificial substrate):
- 10 % peat, air dried and finely ground (2 mm)
- 20 % kaolin clay, kaolinite content > 30 %
- 69 % air dried quartz sand (sand with > 50 % particles
size of 0.05 to 0.20 mm)
- 0.60 % CaCO3
- Organic carbon (%): 10 % peat
- Maximum water holding capacity (in % dry weigth): 72
- CEC: not measured
- Pretreatment of soil: Six days before starting the test the artificial soil was pre-moistened by adding enough deionised water to get 30 % of the maximal water holding capacity of the artificial soil. Until the start of the test, evaporation of the artificial soil was avoided by firmly closing the container.
- Storage (condition, duration): 6 days, under test conditions, darkness


OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark
- Light intensity: 400 - 800 lx


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Adults were counted twice on day 28 (1.0 and 2.5 hours after extraction) to assess later appeared adults, too. The highest counted number of adults was taken into evaluation. Missing adult springtails were counted as dead due to rapid degradation. On day 29, photos of the water surface were prepared using digital photographic equipment. On the resulting print of the respective digital picture juvenile springtails were counted.


VEHICLE CONTROL PERFORMED: yes


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study:Yes
Preliminary Study with C20/22 ATQ trocken: Adult Mortality [%] after 14 Days of
Exposure

Concentration

[mg/kg DW1)] Mortality of adults after 14 days [%] Corrected
mortality
[%]
1. Repl. 2. Repl. 3. Repl. 4. Repl. 5. Repl. Mean
10 0 10 0 0 0 2 0
100 0 0 0 0 10 2 0
1000 0 10 0 10 10 6 4
Control 10 0 0 0 0 2 ¿

Nominal and measured concentrations:

nominal concentrations: 62.5 - 125 - 250 - 500 - 1000 mg/kg DW

Reference substance (positive control):

yes

Remarks:

Boric Acid

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

and reproduction

Remarks on result:

other: n.a.

Effect concentrations 2

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: reproduction, mortality

Remarks on result:

other: n.a.

Effect concentrations 3

Duration:

28 d

Dose descriptor:

LD50

Effect conc.:

> 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: n.a.

Details on results:

- Mortality at end of exposure period: The NOEC was determined to be 500 mg/kg DW, the LOEC was determined to be 1000 mg/kg DW for adult mortality. The mean lethal dose (LD50) was determined to be > 1000 mg/kg soil DW. For exact data please refer to the respective tables.
- No. of offspring produced: The NOEC was determined to be 500 mg/kg DW, the LOEC was determined to be 1000 mg/kg DW for reproduction. The mean effect concentration for inhibition of reproduction (EC50) was determined to be > 1000 mg/kg soil DW. For exact data please refer to the respective tables.
- Other biological observations: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels:
Reference item Boric Acid
Test system Folsomia candida
Exposure Artificial soil
Adult mortality Reproduction
LOEC [mg/kg DW] 100 100
NOEC [mg/kg DW] 50 50
LD50 / EC50 (95 % CI) 120.3 (99.5 ¿ 145.5) 93.6 (88.5 ¿ 98.9)*

Reported statistics and error estimates:

Significant differences in number of adult springtails (mortality) and reproduction were determined in comparison with the control group using appropriate statistical procedures (ANOVA). When running the test a Normality Test and an Equal Variance Test were done first. P-values for both Normality and Equal Variance Test were 0.05.

LD50-value and EC50-value were calculated by sigmoidal dose- response regression. Calculation of the confidence interval for
LD50/EC50-value was carried out using standard procedures. Due to an extrapolation, the LD50-value was not reported.

Any other information on results incl. tables

Number of Alive and Dead Springtails after 28 Days of Exposure

Test item [mg/kg DW]	Replicate
	1		2		3		4		5		6		7		8
	a	d	a	d	a	d	a	d	a	d	a	d	a	d	a	d
62.5	7	3	10	0	8	2	9	1	9	1	-
125	6	4	7	3	9	1	10	0	10	0
250	7	3	9	1	6	4	8	2	9	1
500	7	3	8	2	8	2	7	3	9	1
1000	6	4	5	5	7	3	6	4	6	4
Solvent Control	10	0	8	2	6	4	10	0	8	2	9	1	10	0	10	0
Control	10	0	7	3	10	0	10	0	8	2	6	4	7	3	9	1

a    =    Springtails alive

d    =    Springtails dead

-)    =    Not tested

Adult Mortality [%] after 28 Days of Exposure and Statistics

Test item [mg/kg DW]	Replicate								Mortality
	1	2	3	4	5	6	7	8	Mean ± SD	Corrected*
62.5	30	0	20	10	10	-			14.0 ± 11.4	0.29
125	40	30	10	0	0				16.0 ± 18.2	2.61
250	30	10	40	20	10				22.0 ± 13.0	9.57
500	30	20	20	30	10				22.0 ± 8.37	9.57
1000	40	50	30	40	40				40.0 ± 7.07^1)	30.4
Solvent Control	0	20	40	0	20	10	0	0	11.3 ± 14.6	-
Control	0	30	0	0	20	40	30	10	16.3 ± 16.0	-

SD =    Standard deviation

-)    =    Not tested        *)  =  Values were corrected by pooled control mortality                 

1)   =    Statistically significant different compared with control (ANOVA, p 0.05)

Reproduction of Springtails after 28 Days of Exposure

                               

Test item [mg/kg DW]	Number of juveniles Replicate								Reproduction rate
									Mean + SD	CV [%]	Inhibition* relative to control [%]
	1	2	3	4	5	6	7	8
62.5	351	655	391	399	523	-			464 ± 125	26.9	-7.45
125	153	570	344	445	406				384 ± 153	39.9	11.1
250	510	322	205	490	368				379 ± 126	33.1	12.2
500	479	461	563	452	447				480 ± 48	9.90	-11.3
1000	116	273	268	290	305				250 ± 77^1)	30.6	42.0
Solvent Control	494	376	373	317	245	593	512	670	448 ± 144	32.2	-
Control	396	427	534	390	402	415	387	375	416 ± 50	12.1	-

SD =    Standard deviation

CV =    Coefficient of variation          

-)    =    Not tested                                  *)  =  Calculation based on reproduction data from pooled control                                                          

1)   =    Statistically significant different compared with control (ANOVA, p 0.05)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The No Observed Effect Concentration (NOEC) for mortality and reproduction was determined to be 500 mg/kg DW, the LOEC was determined to be 1000 mg/kg DW for adult mortality as well as for reproduction. The mean lethal and sub-lethal dose (LD50) was determined to be > 1000 mg/kg soil DW.

Executive summary:

The effects of C20/22 ATQ trocken on the reproduction of Folsomia candida in artificial soil were examined in a laboratory study. The study was carried out based on ISO 11267 (1999) and OECD Draft (2008) at the Dr.U.Noack-Laboratorien, Käthe-Paulus-Str. 1, D-31157 Sarstedt, from 2008-04-18 to 2009-07-22.

The aim of the test was to determine the effects of the test item on the reproduction of Folsomia candida in artificial soil by cutaneous and alimentary uptake. The test item C20/22 ATQ trocken was incorporated once into artificial soil with the test item concentrations 62.5 - 125 - 250 - 500 - 1000 mg/kg DW. The control was artificial soil without the test or reference item. Additionally a solvent control was tested. Folsomia candida (12 days old) were placed into artificial soil. For the control and solvent control 8 replicates and for the test item 5 replicates were tested. In each replicate 10 juveniles were inserted. During exposure springtails were fed with granulated dry yeast. After 28 days, mortality and reproduction were assessed.

Toxicity to Folsomia candida, Laboratory Test

Test item	C20/22 ATQ trocken
Test object	Folsomia candida
Exposure	Artificial soil
	Adult mortality	Reproduction
LOEC [mg/kg DW]	1000	1000
NOEC [mg/kg DW]	500	500
LD50 / EC50 [mg/kg DW]	> 1000	> 1000

DW = Dry weight                                            CI = Confidence Interval

The validity of the study was evaluated according to ISO 11267 (1999) and OECD Draft (2008). Control mortality should be 20 %. In this study the mean control mortality (pooled controls) was 13.8 %. The reproduction rate should be > 100 instars per control replicate. In this study the reproduction rate was between 245 and 670 instars per control replicate (pooled control). The coefficient of variation of the reproduction rate in the control should be 30.0 %. In this study the coefficient of variation in the pooled controls was 25 %. Thus, this study was considered to be valid.