Phenoxymethylpenicillin potassium

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 September 2019 - 5 November 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Test Item name: Pen V Potassium
Lot No.: B519322
Appearance: white, solid powder
Expiry date: 30 April 2024
Storage: room temperature, protected from light

Analytical monitoring:

yes

Details on sampling:

For the determination of the test item concentration, samples were taken from the test item treated group and from the control group (4 x 5 mL at the start and 4 x 5 mL at the end of the study).

Vehicle:

no

Details on test solutions:

The test solution used in the test was prepared by mechanical dispersion without using any solubilizing agent. An amount of 0.0444 g test item was dissolved in 444 mL dilution water (ISO Medium, see 5.4) in order to obtain the nominal concentration of 100 mg/L. The test solution was freshly prepared in the testing laboratory just before introduction of test animals (start of the experiment).

Test organisms (species):

Daphnia magna

Details on test organisms:

Species and strain: Daphnia magna (Straus)
Origin: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
Breeding: The Daphnia are bred in the Laboratory of TOXI-COOP ZRT.
Number of animals: Twenty animals in both groups (concentration and untreated control), divided into 4 concurrent batches (5 animals per batch).
Age of animals: Less than 24 h old at the beginning of the test.
Sex: Female
Animal health: Apparently healthy animals were used in this test with a known history
Acclimatization: Test animals were bred under similar (or the same) conditions as that used during the exposure period (holding water, temperature, background colour etc.), therefore additional acclimatisation before the test was not necessary. Brood daphnids were maintained in dilution water at the test temperature for at least 48 hours prior to the start of the test.
Food and Feeding: The Daphnia were fed with concentrated algal suspension of Raphidocelis subcapitata during the holding. Test animals were not fed during the exposure.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

According to Guidelines.

Post exposure observation period:

No.

Hardness:

The reconstituted water (ISO medium) had an approximate theoretical total hardness of 249 mg/L (as CaCO3).

Test temperature:

The test temperature was between 18 °C and 22 °C with a maximum deviation of ± 1 °C. As minimum, the ambient temperature was measured continuously with a min/max thermometer in the climate chamber. Water temperature was additionally determined in test vessels at the start and at the end of the test.

pH:

The pH did not vary by more than 1.5 units in any one test and was within the range of 6 - 9.

Dissolved oxygen:

The dissolved oxygen concentration and pH were measured in the control and in the test concentration at the start and at the end of the experiment. The oxygen concentration was ≥ 3 mg/L at the temperature used.

Salinity:

No.

Conductivity:

Not reported.

Nominal and measured concentrations:

0 - 100 mg/L nominal. The measured concentrations varied between 80 and 101 % of the nominal.

Details on test conditions:

The test vessels (glass beaker; volume approx. 50 mL filled with approx. 40 mL control or test solution) were kept in a climate chamber under controlled environmental conditions during the test and test solutions were not aerated. The water temperature, dissolved oxygen concentration and pH were measured in each test group at the start (before test solutions had been distributed into the test vessels) and in each test vessel at the end of the test. Additionally, the ambient temperature was measured continuously using a min/max thermometer in the climate chamber.

Test type:Static test. Based on the results obtained during analytical method validation (Study number: 880-100-4606) the test item is stable for the duration of 48 hours in ISO Medium, therefore, a static test was performed.

Temperature: The test temperature was between 18 °C and 22 °C with a maximum deviation of ± 1 °C. As minimum, the ambient temperature was measured continuously with a min/max thermometer in the climate chamber. Water temperature was additionally determined in test vessels at the start and at the end of the test.

Oxygen Concentration and pH: The dissolved oxygen concentration and pH were measured in the control and in the test concentration at the start and at the end of the experiment. The oxygen concentration was ≥ 3 mg/L at the temperature used. The pH did not vary by more than 1.5 units in any one test and was within the range of 6 - 9.

Light-Dark Cycle: 16 h light : 8 h dark. Hardness: The reconstituted water (ISO Medium) has an approximate theoretical total hardness of 249 mg/L (as CaCO3).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate.

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Analytical Results
The quantification of the test item in test solutions was performed according to a previously validated analytical method (Study Code: 880-100-4606) by the analytical laboratory of TOXI-COOP ZRT. Samples were taken from the test concentration and from the control at the start and at the end of the experiment and analysed by HPLC method with UV detection. The mean of the measured concentrations of the test item in the test solution was 96 % of the nominal value at the start and 86 % of the nominal value at the end of the test. There is evidence that the concentration of the tested substance was maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on the nominal concentration value. Test item was not detected in the control group.

Biological Results
Biological results are based on the nominal test item concentration. No immobilisation or any abnormal behaviour of the test animals was observed in the control group and no immobilization or any abnormal behavior of the test animals was observed at the test concentration of 100 mg/L after 48 hours of exposure.

Results with reference substance (positive control):

The date of the last study (Study Number: 392-202-4466) with reference item Potassium dichromate was: 12 - 13 March 2019. The 24-h EC50: 1.58 mg/L (95 % conf. limits: 1.34 – 1.85 mg/L)

Validity of the Study

Immobilisation was not observed in the control group and the dissolved oxygen concentration in control and test vessels was in the range of 8.16 – 8.57 mg/L during the experiment (more than 3 mg/L in all cases).

All validity criteria were within acceptable limits and, therefore, the study can be considered as valid.

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity test on Daphnia magna, the obtained results showed that the test item Pen V Potassium had no toxic effects on the mobility of Daphnia up to the test concentration of 100 mg/L. The 48-h LOEC was determined to be higher than 100 mg/L and the 48-h NOEC was determined to be 100 mg/L (directly from the raw data).The results are based on the nominal test item concentration (100 mg/L).

All validity criteria were met and, therefore, the study can be considered as valid.

Executive summary:

The test substance Pen V Potassium was assessed using Daphnia magna in a 48-hour Acute Immobilisation Test. Young Daphnia were exposed to aqueous test media containing the test item for 48 hours. Based on results obtained in a non-GLP preliminary experiment Daphnia were exposed to a single concentration of 100 mg/L (limit concentration). A concurrent control group was run. A static test was performed as the test item was previously shown to be stable in the ISO Medium for 48 hours. The analytically measured test item concentration remained within ± 20 % of the nominal during the test period of 48 hours; therefore, biological results are based on the nominal concentration. Healthy, young female daphnids with known origin, history and acclimatisation conditions were used. Twenty Daphnia(divided into 4 replicates) were tested in each test group, exposed to 100 mg/L test item or run as control. Each test vessel contained approximately 40 mL test solution. Fully characterised (content, physico-chemical characteristics) ISO Medium was used as test medium. The immobilisation of the test animals was recorded 24 and 48 hours after treatment. Environmental conditions were recorded. All measured values remained within acceptable ranges. No immobilisation or any abnormal behaviour of the test animals was observed in the control group and no immobilization or any abnormal behavior of the test animals was observed at the test concentration of 100 mg/L after 48 hours of exposure. The 48-h NOEC was determined to be 100 mg/L and the 48-h LOEC was determined to be > 100 mg/L (directly from the raw data). All validity criteria were met and, therefore, the study can be considered as valid.

Biological endpoints are summarised below:

Endpoints	Concentration [mg/L] based on nominal concentration
48-h NOEC	100
48-h LOEC	> 100

 

Description of key information

A static 48h-test with Daphnia magna was performed. The test item had no toxic effect at 100 mg/L concentration (i.e. limit test concentration) on Daphnia magna; the EC10, EC20, EC50 results and the LOEC are higher than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The EC50 is not 100 mg/L, as one is forced to enter in the field above, but the EC50 is >100 mg/L.