1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylindeno[5,6-c]pyran

Toxicity to terrestrial arthropods

Administrative data

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24/09/1996 -25/08/1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Reliability 2 is assigned because the test has been carried out following internation guideling and under GLP without deviations that influence the results, however, the tested concentrations were not verified with expertimental analysis

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report Date:

1997

Materials and methods

Test guideline

Qualifier:

according to

Guideline:

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

Deviations:

yes

Remarks:

: ISO/CD 11267 ISO Draft Soil Quality: Effects of soil pollutants on Collembola (Folsomia candida): Mathod for the determination of effects on reproduction (Version 3/96)

GLP compliance:

yes (incl. certificate)

Application method:

soil

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

Batch # 7953-I

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No data

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

Samples were taken at the start of the test (7 days after application), first day of exposure and at the end of the study.

Test substrate

Vehicle:

yes

Details on preparation and application of test substrate:

Acetone was used as a solvent.

Test organisms

Test organisms (species):

Folsomia candida

Animal group:

Collembola (soil-dwelling springtail)

Details on test organisms:

The test was started with juvenile springtails of the species Folsomia candida 10 to 12 days of age and survival and reproduction after 28 days were determined.

Study design

Study type:

laboratory study

Limit test:

yes

Total exposure duration:

28 d

Post exposure observation period:

No data

Test conditions

Test temperature:

17 - 25 degrees Celsius

pH (if soil or dung study):

at start of study pH 6.1 - 6.4, after 4 weeks pH 5.7 - 5.9

Humidity:

water content adjusted to 40-60% of total water holding capacity, corresponding to approx. 30% of dry weight.
Water content at start of study: 33.6 - 34.9 % of dry weight, after 4 weeks: 26.6 - 30.6 % of dry weight

Photoperiod and lighting:

dark/light cycle 8h/16h, 400 - 530 lux

Details on test conditions:

Composition artifical soil: 10% sphagnum-peat; 20% kaolinite clay;circa 0.5% CaCO3 to adjust pH to 6.0+/- 0.5; approx. 50.1% fine quartz sand (grain size 0.1 - 0.5 mm). Solvent: acetone; test range: 1 - 105 mg/kg.
Animals were fed with granulated dry yeast, added during the test when necessary.
The test materials at appropriate concentrations were dissolved in equal amounts of acetone, mixed with the quartz sand and allowed to slowly evaporate and mixed with the standard (DIN) soil. After preparation of the test concentrations and an equilibrium period of one week, the test organisms were added to the soil. The test medium was not refreshed during the test period.
10 springtails per test container. Test containers were covered with a lid to prevent excessive evaporation. Opening for 10 seconds twice a week for aeration. Water (after weighing) and food added when needed.

Nominal and measured concentrations:

Nominal test concentrations were 1, 3, 8, 19, 45 and 105 mg/kg soil. Samples were taken in case analyses would be considered to be necessary. No analyses were carried out.

Reference substance (positive control):

yes

Remarks:

Betanal (CQ 532); active ingredient 160g/l Phenmdeipham. Applied 200 mg Betanal/kg soil.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

4 wk

Dose descriptor:

NOEC

Effect conc.:

45 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

28 d

Dose descriptor:

LOEC

Effect conc.:

105 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

No significant mortality was observed in soils containing up to 45 mg/kg, but mortality was 72% in 105 mg/kg. Reproduction was inhibited by 23% in 45 mg/kg. However, this inhibition was not statistically significantly different from the control. The reproduction in 105 mg/kg was 16% of the control. Thus, the NOEC for HHCB is 45 mg/kg based on survival

Results with reference substance (positive control):

No mortality was observed for the reference compound and the reproduction was 56.3% of the controls.

Reported statistics and error estimates:

Mortality: Significance of acute toxic effects were tested with Mann&Whitney U-test (p = 0.05); effects at a dosage of 105mg/kgdw (artificial soil) differed significantly from the control.
Reproduction: reproduction rates at the highest dose tested (105 mg/kgdw) were significantly reduced (by 84%) to the control (Dunnett test, p = 0.05)

Any other information on results incl. tables

Table - Mortality of adult springtails per replicate (mean per treatment, standard deviation, n = 5)

Treatment	Concentration (mg/kgdw)	Mean (% of initial number))	Std.dev. (%)
control	0	0.0	0.0
solvent control	0	2.0	3.3
C1	1.0	0.0	0.0
C2	3.0	4.0	5.0
C3	8.0	0.0	0.0
C4	19.0	6.0	4.4
C5	45.0	4.0	4.4
C6	105.0	72.0	25.9
reference compound		0.0	0.0

 

Table - Reproduction: number of juvenile springtails per replicate (mean per treatment, standard deviation, n = 5)

Treatment	Concentration (mg/kgdw)	Mean	Std.dev.
control	0	395
solvent control	0	338	82
C1	1.0	466	72
C2	3.0	409	101
C3	8.0	380	119
C4	19.0	490	83
C5	45.0	284	87
C6	105.0	59	17
reference compound		207	55

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Mortality was 1% (required < 20%); mean reproduction rate 367 per control vessel (required at least 100), with coeff. of var. 22% (required < 30%).

Conclusions:

4wk-NOEC = 45 mg/kg dwt; LOEC = 105 mg/kg dwt.

Executive summary:

The springtail Folsomia candida was tested according to the draft ISO/CD 11267. The test materials at appropriate concentrations were dissolved in equal amounts of acetone, mixed with the quartz sand and allowed to slowly evaporate and mixed with the standard soil containing 10% Sphagnum peat, 20% kaolinite clay, approximately 70% fine quartz-sand (grain size 0.1-0.5 mm) and 0.5% calcium carbonate to adjust to pH 6.0±0.5. After preparation of the test concentrations and an equilibrium period of one week, the test organisms were added to the soil. The test medium was not refreshed during the test period.

The test was started with juvenile springtails of the species Folsomia candida 10 to 12 days of age and survival and reproduction after 28 days were determined. Nominal test concentrations were 1, 3, 8, 19, 45 and 105 mg/kg soil. The animals were fed with granulated dry yeast. No significant mortality was observed in soils containing up to 45 mg/kg, but mortality was 72% in 105 mg/kg. Reproduction was inhibited by 23% in 45 mg/kg. However, this inhibition was not statistically significantly different from the control. The reproduction in 105 mg/kg was 16% of the control. Thus, the NOEC for HHCB is 45 mg/kg based on survival.