Carbamic acid, [5-isocyanato-2(or 4)-methylphenyl]-, C10-14-alkyl esters

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 26, 2001 until December 7, 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to OECD Guideline 203 (Fish, Acute Toxicity Test) under GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

440-930-8

EC Name:

-

Cas Number:

330198-48-6

Molecular formula:

C19H28N2O3 / C21H32N2O3 / C23H36N2O3 / C29H50N2O4 / C31H54N2O4 / C33H58N2O4 / C35H62N2O4 / C37H66N2O4

IUPAC Name:

Condensation products of 4-methyl-m-phenylene diisocyanate with alcohols, C10-14 (even numbered)

Details on test material:

Identity: ZP-TIX 1014
Batch: NW-01-028
Purity > 99%,
Appearance: Solid
Expiration date: 30.12.2001
Storage: At room temperature at about 20°C, in the dark
Color: white (visual judgement by RCC)
Stability in water: Determined in RCC Study 818897
Stability under test conditions: Not stable in test water, the degradation products were tested
Solubility in water: Insoluble (sponsor information) determined in RCC Study 818651

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
Common name: Zebra-fish
Strain: Brachydanio rerio
Source: Aquaruimcenter Wildenauer, Munich, Germany
Length at study initiation (length definition, mean, range and SD): 3.0 +/- 0.2 cm
Weight at study initiation (mean and range, SD): 0.28 +/- 0.05 g
Feeding before test: until one day before start of the test
Food type: comercial fish diet "Tetra Min Hauptfutter"
Source: TETRA-Werke, D-49304 Melle, Germany

ACCLIMATION
Acclimation period: one week
Acclimation conditions: same as test
Type and amount of food: comercial fish diet "Tetra Min Hauptfutter" (source see above)
Feeding frequency: in accordance with guidelines until one day before start of the test
Health during acclimation (any mortality observed): all fish were healthy

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

none

Test conditions

Hardness:

Before start of the test: 2.5 mmol/L (= 250 mg/L) as CaCO3
During test: no data

Test temperature:

From 21 to 22 °C

pH:

From 7.8 to 8.0

Dissolved oxygen:

From 7.4 to 8.1 mg/L

Salinity:

CaCl2 x 2H2O 2.0 mmol/L (= 294 mg/L)
MgSO4 x 7H20 0.5 mmol/L (= 123 mg/L)
NaHCO3 0.75 mmol/L (= 65 mg/L)
KCl 0.075 mmol/L (= 5.8 mg/L)

Nominal and measured concentrations:

100 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: aquarium (1 aquarium for each treatment)
- Material, size, headspace, fill volume: 20 L glass
- Aeration: slightly during the test
No. of organisms per vessel: 7
No. of vessels per concentration (replicates): 1
No. of vessels per control (replicates): 1
Biomass loading rate: 0.2 g fish wet weight per liter test medium

TEST MEDIUM / WATER PARAMETERS
Source/preparation of dilution water: Reconstituted water: analytical grade salts were dissolved in deionized water to obtain the following nominal concentrations
Alkalinity: 0.8 mmol/L
Ca/Mg ratio: 4:1 (based on mortality)
Na/K ratio: 10:1 (based on mortality)
Culture medium different from test medium: no
Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
Adjustment of pH: no
Photoperiod: A 16-hour light to 8-hour darkness photoperiod with a 30 minute transition period
Light intensity: within the range of 50 to 500 LUX

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Fish: observation after approximately 2, 24, 48, 72 and 96 h test duration for mortality, and visible abnormalities
Water: measurements of temperature, pH-values, dissolved oxigen concentrations at the start of the test and daily during test period
Appearance of the test medium: daily recorded

TEST CONCENTRATIONS
Spacing factor for test concentrations: none (only one concentration; limit test)
- Test concentrations: 100 mg/L

ANALYSIS OF TEST ITEM CONCENTRATION
The test item is a poorly soluble mixture of isocyanates and it is generally known, that these compounds degrade fast in the test medium (according to pre-experiments without GLP. Furthermore, the verification of the test item could not be based on its degradation products, since none could be defined during the analytical pre-experiment. For this reasons, no analytical work was performed.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Behavioural abnormalities: none
Observations on body length and weight: no data
Other biological observations: no
Mortality of control: 0
Other adverse effects control: no
Abnormal responses: no
Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
Effect concentrations exceeding solubility of substance in test medium: yes
Behavioural abnormalities: no abnormalities
Observations on body length and weight: no data
Mortality of control: 0
Other adverse effects control: none
Abnormal responses: no data
Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no data

Reported statistics and error estimates:

no data

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the controls and at the loading rate of 100 mg/L no mortality or visible abnormalities were determined at the test fish during the test period of 96 hours.

Executive summary:

The acute toxicity of the test item ZP- TIX 1014 to zebra fish(Brachydanio rario) was determined in a 96-hour static test according to the EU Commission Directive 92/69/EEC, Part C.1 (1992), and the OECD Guideline for Testing of Chemicals No. 203, (1992). The test method is also based on the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures,2000.

 

In order to assess the toxicity of ZP-TIX 1014 to fish, a water accommodated fraction (WAF) with a loading rate of 100 mg/L was tested. Additionally, a control (test water without test item) was tested in parallel. Thus, a limit test was performed in accordance with the EU Commission Directive 92/69/EEC to demonstrate that the test item has no toxic effect on the fish up to the loading rate of 100 mg/L.

 

The W AF with the loading rate of 100mg/Lwas prepared by dispersing the test item in test water by stirring for 96 hours. Then, the dispersion of the test item was filtered through a membrane filter (pore size 0.45 µm). The undiluted filtrate of the dispersion was used as test medium.

 

The test item is a poorly soluble mixture of isocyanates and it is generally known, that these compounds degrade fast in water. Due to these characteristics, the test item could not be analytically measured in the test medium. Furthermore, the verification of the test item could not be based on its degradation products, since none could be defined during the analytical pre-experiments. For this reasons, no analytical work was performed.

Since the test item is not stable in test water, the degradation products were tested. Therefore, a long stirring period of 96 hours was chosen to enrich the test water with the maximum concentration of degradation products within a practically acceptable time period.

 

The biological test results after 96 hours test duration.

 

In the control and at the loading rate of 100mg/Lnomortality or visible abnormalities were determined at the test fish during the test period of 96 hours. Therefore, the 96-hour NOEC (highest concentration tested without toxic effects after the exposure period of 96 hours), and the 96-hour LC0 of the test item ZP-TIX 1014 and its degradation products, respectively, to zebra fish were determined to be at the loading rate of 100 mg/L.

The NOEC and the LC0 might even be higher, but loading rates in excess of 100mg/L have not been tested, according to the guidelines.

The 96-hour LOEC (lowest concentration with toxic effects), the 96-hour LC50 and the 96-hour LC100 were clearly higher than the loading rate of 100mg/L.These values could not be quantified due to the absence of toxicity of ZP-TIX 1014 at the tested loading rate.