(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-enecarboxylic acid

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted under GLP conditions and based on the ISO International Standard.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-enecarboxylic acid

EC Number:

205-334-2

EC Name:

(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-enecarboxylic acid

Cas Number:

138-59-0

Molecular formula:

C7H10O5

IUPAC Name:

(3R,4S,5R)-3,4,5-trihydroxycyclohex-1-enecarboxylic acid

Constituent 2

Reference substance name:

Shikimic acid

IUPAC Name:

Shikimic acid

Details on test material:

- description : white powder

Range-finding test
- Purity : 98,9%
- Test substance storage : in refrigerator in the dark
- Stability under storage conditions : stable
- Expiry date : 3 March 2002

Final test
- Purity : 100,4%
- Test substance storage : in refrigerator in the dark
- Stability under storage conditions : stable
- Expiry date : 31 December 2001

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Species : Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Reason for selection : this system has been selected as an internationally accepted species.
- Validity of batch : frequent inspection of the culture with respect to number of young, appearance of young and parental daphnia and global feeding behaviour
- Characteristics : for the test selection of young daphnia with an age of < 24 hours.

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

Temperature of medium : daily in all vessels, beginning at the start of the test.

pH:

At the beginning, after 24 h and at the end of the test, for all concentrations and the control.

Details on test conditions:

The project started with a static range-finding test exposing Daphnia to Shikimic Acid concentrations of 0.1, 1.0, 10 and 100 mg/l. No daphnids became immobilised during the 48-hour test period up to and including 10mg/l, while 60% of the daphnids exposed to 100 mg/l became immobilised. Hence, the 48h-EC50 was expected to be between nominally 10 and 100 mg/l. Analysis of samples taken during the range-finding test of the study with Carp (NOTOX project 322289) showed that a shikimic acid concentration of 100 mg/l decreased by more than 20% during the first 24 hours of exposure. As a consequence of the instability in the test medium further testing was performed applying continuous renewal of test solutions in a flow-through system.

Reference substance (positive control):

yes

Remarks:

Test medium without test substance or other additives.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

116 mg/L

Nominal / measured:

meas. (not specified)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 10 - <= 100 mg/L

Nominal / measured:

nominal

Details on results:

In the controls, no Daphnia had been immobilised or trapped at the surface of the water.

The analytical program confirmed that the actual concentrations had been maintained at the target concentration +/- 20 %. The only exception was the measured concentration at the end of the test at 18 mg/l. However, this did not affect the results of this study.

Further, all test conditions (pH, dissolved oxygen and temperature) remained within the ranges prescribed by the protocol.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Acute Toxicity Study in Daphnia Magna with shikimic acid.
The study procedure described in this report were based on the ISO International Standard 6341 : "Water quality - Determination of the inhibition of the mobility of Daphnia magna Straus. Acute toxicity test, Third edition, 1996-04-01".

Executive summary:

The project started with a static range-finding test exposing Daphnia to Shikimic Acid

concentrations of 0.1, 1.0, 10 and 100 mg/I..No daphnids became immobilised during the 48-hour test

period up to and including 10 mg/1, while 60% of the daphnids exposed to 100 mg/1 became

immobilised. Hence, the 48h-ECs0,was expected to be

between nominally 10 and 100 mg/1. Analysis of samples taken during the range-finding

test of the study with Carp (NOTOX Project 322289) showed that a Shikimic Acid concentration of 100

mg/1 decreased by more than 20% during the first 24 hours of exposure. As a consequence of the

instability in the test medium further testing was performed applying continuous renewal of test

solutions in a flow-through system. The project was continued with an EC50,study under flow-through

test conditions with continuous renewal of test media at target concentrations between 10 and 100

mg/1 with an increasing factor of 1.8. The test was performed in duplicate with 10 Daphnia per

vessel. Exact volumes of a 20 mg/ml stock solution in Milli-RO water were dosed directly by 5

Gilson Dispensers via a computer-controlled system. The dosed volumes entered a mixing flask separately from the tap water supply. The tap water was supplied via a flowmeter with a flow rate of ca. 12 L/h. In the mixing flasks,the dosed volumes and the tap water were mixed under continuous stirring. The whole system was checked daily. Analysis of the samples taken

during the final test showed that the measured concentrations had been maintained at the target concentrations

Shikimic Acid did not induce significant acute immobilisation of Daphnia magna at or below a mean

measured exposure concentration of 116 mg/1 under flow-through test conditions (NOEC).