Carbamic acid, N-[(1R)-2-hydroxy-1-phenylethyl]-, 1,1-dimethylethyl ester

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:

April 20, 2016 to Dec 6, 2016

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

In range-finding test, the concentrations were chosen to be 1.00, 10.0, 100 and 150 mg/L which deviated from protocol the concentrations of 1.00, 10.0 and 100 mg/L. However, the test results showed that the minimum concentration causing 100% mortality

GLP compliance:

yes (incl. QA statement)

Remarks:

This experiment strictly complied with "The guidelines of chemical testing good laboratory practices (HJ/T 155)'', CMA (China Metrology Accreditation) and CNAS (China National Accreditation Service for Conformity Assessment).

Test material

Test material information

Constituent 1

Reference substance name:

Carbamic acid, N-[(1R)-2-hydroxy-1-phenylethyl]-, 1,1-dimethylethyl ester

EC Number:

600-273-1

Cas Number:

102089-74-7

Molecular formula:

C13H19NO3

IUPAC Name:

Carbamic acid, N-[(1R)-2-hydroxy-1-phenylethyl]-, 1,1-dimethylethyl ester

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Batch No. : LS-53-20150815
Appearance: Powder
Aqueous solubility 0.45 g/L (25°C)
Storage conditions: Room temperature ( 10-30°C)
Purity / Assay: 99.99%

Sampling and analysis

Analytical monitoring:

yes

Remarks:

The analytical results showed that the concentration of the test substance is consistent in the test medium throughout the 96-hour test period (deviation within 20%), thus a static procedure was reasonable.

Test solutions

Vehicle:

no

Details on test solutions:

In the test, test solutions of N-Boc-D-phenylglycinol had been prepared by directly
dissolving appropriate amount of test substance in dilution medium (Vol: 3L) and then facilitating its
dispersion by stirring for 30 min.

Test organisms

Details on test organisms:

Fish (Gobiocypris rarus)

TEST ORGANISM
- Common name: Rare minnow, Gobiocypris rarus
- Strain: (Batch No. F20160415G)
- Source: Obtained from a fish supplier of Institute of Hydrobiology, Chinese Academy of Sciences.
- Length at study initiation (length definition, mean, range and SD): Mean: 2.78cm. Range:2.69-2.92cm.
- Weight at study initiation (mean and range, SD): Mean: 0.222g Range: 0.202-0.35g

ACCLIMATION
- Acclimation period: Fish were held at least for 12 days in holding tanks supplied with a continuous flow of aerated water before being used for testing. Fish to be used in the test were held for at least 7 days in water of the quality and temperature to be used in the test.
A photoperiod of 16 hours light, provided by overhead fluorescent tubes, and 8 hours dark
was maintained. The oxygen concentration was more than 80% of the air saturation value.

FEEDING DURING TEST
- Food type: The ingredients of the fish food are given as follows:
Crude Protein> 36.0%
Crude .Fat> 2.0%
Crude Fibre< 3.0%
Crude Ash <13.5%
Moisture< 10.0%

- Frequency: The fish was fed daily during the holding period on proprietary fish food. They were held without food for approximately 24 hours before being placed into the test vessels.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

The test fishes were exposed for 96 hours to the test solution. The study comprised at least one range-finding test followed by a definitive test to determine the 96-hour median lethal concentration (LC50) and 95% confidence limit of the test substance.

Test conditions

Hardness:

The total hardness was in the range of 157 to 168 mg (CaC03)/L.

Test temperature:

The temperature of the test mediums was maintained in the range of 22.8°C to 23.2°C

pH:

During the whole test period, the pH values of the control mediums and test mediums were
between 7.76 and 7.99.

Dissolved oxygen:

The Dissolved Oxygen (DO) values varied from 74% ~ 95% of the air saturation.

Nominal and measured concentrations:

Nominal concentrations of 1.00, 10.0, 100 and 150 mg/L were used in the range-finding test. While the nominal concentrations of 10.0, 20.0, 40.0, 80.0 and 150 mg/L were used in the definitive test.
For the definitive test, the measured concentrations were 9.10, 20.0, 39.5, 79.6 and 145 mg/L, respectively.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, K2Cr20 7, CAS: 7778-50-9; Purity: 2: 99.8% Lot number: LVC00524; Beijing Leon Technology Co. Ltd.

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

72.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The results showed that under valid static test conditions, the 96 h-LC50 of the test
substance to fish (Rare minnow, Gobiocypris rarus) was 72.4 mg/L, with 95% confidence
limit of 56.8 mg/L - 92.4 mg/L (based on the measured concentration). The maximum
tested concentration causing no mortality (96 h-LC0) was 20.0 mg/L (measured
concentration 20.0 mg/L). The minimum concentration causing 100% mortality (96 h-LC100) was 150 mg/L (measured concentration 145 mg/L), i.e.:
96 h-LC50= 72.4 mg/L, 95% CI 56.8 mg/L - 92.4 mg/L (based on measured concentration);
96 h-LC0 = 20.0 mg/L (measured concentration 20.0 mg/L);
96 h-LC100= 150 mg/L (measured concentration 145 mg/L).

Executive summary:

Under static conditions, the acute toxicity of test substance (N-Boc-D-phenylglycinol) to Rare minnow (Gobiocypris rarus) was conducted according to: "The guidelines for the testing of chemicals" (HJ/T 153-2004), The Guidelines for the Testing of Chemicals, Effects on Biotoc Systems"(the 2nd edition); and with reference to Procedure 203 of the "Guidelines for Testing of Chemicals" of the OECD: "Fish, Acute Toxicity Test" (1992). A range-finding test and then a definitive test were performed respectively. Nominal concentrations of 1.00, 10.0, 100 and 150 mg/L were used in the range-finding test. While the nominal concentrations of 10.0, 20.0, 40.0, 80.0 and 150 mg/L were used in the definitive test. Water samples taken from the blank control and the treatments in the definitive test were analysed. Concentrations of the test substance were quantified using
UPLC-PDA of the measured concentrations were 9.10, 20.0, 39.5, 79.6 and 145 mg/L, respectively. The analytical results showed that the concentration of the test substance is consistent in the test medium throughout the 96-hour test period (deviation within 20% ). Thus a static procedure was reasonable. In the range-finding test, 5 fishes per treatment and no replicates were used. And 10 fishes per treatment of no replicates were used in the definitive test. The test fishes were exposed for 96 hours to the test solution. All validity criteria were met, therefore the test was considered valid.
During the test period, all fish in the control and treatment of 10.0, 20.0 mg/L were alive and appeared normal. Effects (Fish lying on side or back) occurred at concentrations of 40.0 mg/Land above. All Fish were dead after 96 h-exposure at concentration of 150 mg/L. The results showed that under valid static test conditions, the 96 h-LC50 of the test substance to fish (Rare minnow, Gobiocypris rarus) was 72.4 mg/L, with 95% confidence limit of 56.8 mg/L ~
92.4 mg/L (based on the measured concentration). The maximum
tested concentration causing no mortality (96 h-LC0) was 20.0 mg/L (measured
concentration 20.0 mg/L). The minimum concentration causing 100% mortality (96h-LC100) is 150 mg/L (measured concentration 145 mg/L), i.e.:
96 h-LC50 = 72.4 mg/L, 95% CI 56.8 mg/L ~ 92.4 mg/L (based on measured concentration);  96 h-LC0 = 20.0 mg/L (measured concentration 20.0 mg/L);
96 h-LC100 = 150 mg/L (measured concentration 145 mg/L).