Phenoxymethylpenicillin potassium

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

9 September 2019 - 05 November 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

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:

no

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Origin:
Akvárium Magazin Kft. (Pasaréti Gyula)1222 Budapest, Dévény u. 36.
Number of animals:
There were 10 animals in the test item treated and in the control group respectively (no replicates were used).
Age and size of animals:
Juveniles were used. The body length was within a range of 1-2 cm. The total body weight of the test animals per groups was recorded on the day 0 (at the start of the test). The loading of the test aquaria was calculated (g fish/liter test solution). The loading of the test aquaria did not exceed 0.8 g / L.
Sex:
Both female and male (were not separated).
Animal health:
Apparently healthy animals were used in the test.

Acclimatization, holding of fish prior to the study:
Fish were held for at least 9 days before test initiation in the fish laboratory of TOXI-COOP ZRT. under the same conditions as used during the exposure period. During holding, fish were fed with appropriate, commercial diet for fish at least three times per week until 24-48 hours before the test start. The health of the breeding was continuously monitored and any mortality or abnormal behaviour recorded. No mortality (less than 5 % of population) occurred in seven days before the start of the experiment, therefore the batch was considered to be acceptable for testing.
Photoperiod:
12 to 16 hours light daily;
Temperature:
21 – 25°C; constant within ± 2 °C;
Oxygen concentration:
at least 80 % of air saturation;
Feeding:
three times per week or daily until 24-48 hours before the test is started.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

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

Test temperature:

Aquariums were kept in a climatic chamber, maintained at a temperature in the range of 21-25 °C, constant within ± 2 °C.

pH:

The preferable range of pH for control is between 6.0 and 8.5. The test was carried out without adjustment of pH.

Dissolved oxygen:

Not less than 60 % of the air saturation value.

Salinity:

No.

Nominal and measured concentrations:

100 mg/L (limit concentration)

Details on test conditions:

In order to select appropriate test concentrations for use in the definitive test, two non-GLP preliminary range-finding tests were conducted to determine the approximate toxicity of the test item. In the first static preliminary test three fish were exposed for 96 hours in each test group (no replicates were used). A control group ran concurrently. For preparation of the test solution an amount of 0.1008 g test item was dissolved in 1008 mL ISO Medium to obtain the 100 mg/L nominal concentration.
In the second semi-static preliminary test three fish were exposed for 96 hours in each test group (no replicates were used). A control group ran concurrently. For preparation of the test solution an amount of 0.1005 g test item was dissolved in 1005 mL ISO Medium to obtain the 100 mg/L nominal concentration in the first renewal period and an amount of 0.1006 g test item was dissolved in 1006 mL ISO Medium to obtain the 100 mg/L nominal concentration in the second renewal period.
The preliminary range-finding tests were not performed in compliance with the GLP-Regulations and are excluded from the Statement of Compliance (Statement of the Study Director) in the Final Report, but the raw data of these tests will be archived under the study code of the present study. Based on the results of the non-GLP preliminary range-finding tests, single concentration of 100 mg/L (limit concentration) was tested in a limit test with one renewal on day 2. A concurrent control group ran parallel. The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. Therefore, the biological results are based on the nominal test item concentration.
The test solution used in the test was prepared by mechanical dispersion without using any solubilising agent. For preparation of the test solution an amount of 0.5000 g test item was dissolved in 5000 mL ISO Medium to obtain the 100 mg/L nominal concentration in the first renewal period and an amount of 0.5003 g test item was dissolved in 5003 mL ISO Medium to obtain the 100 mg/L nominal concentration in the second renewal period.
The test solution was freshly prepared in the testing laboratory just before introduction of fish (at the start of each renewal period).

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

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:

There was no mortality observed in 10 fish in the control group and in 10 fish exposed to the test item concentration of 100 mg/L. No sub-lethal effect was observed in the control and in the test item treated group during the study. The 96-h No Observed Effect Concentration (NOEC) was determined to be 100 mg/L. The 96-h and the 96-h LOEC values were determined to be > 100 mg/L based on nominal test concentration.

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity test on Zebrafish (Danio rerio)the test item Pen V Potassium had no toxic effect on fish at the limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be >100 mg/L. The 96-h NOEC value was determined to be 100 mg/L. All validity criteria were met and biological endpoints are given directly from the raw data.

Executive summary:

Fish were exposed to aqueous test media containing the test item for 96 hours test period in a semi-static test. Ten fish were exposed to the test concentration or run as control (no replicates were used). Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily.Endpoints:NOEC, LOEC and LC50 for 96-h test period. Statistics:No mortality or sublethal effects were observed, therefore no statistical analysis was performed. Endpoints were determined directly from the raw data.Validity:All validity criteria were met and therefore the study can be considered as valid (see section 10.1) Analytics:The quantification of the test item in the test solutions was performed according to a previously validated analytical method (Study No.: 880-100-4606). Samples were taken from the test concentration and from the control at the start and at the end of each renewal period and analysed by HPLC method with UV detection. Results:No mortality or sub-lethal effects were observed during the 96-h exposure period in the test item treated or in the control group. The measured test item concentrations remained within ± 20 % of the nominal during the test period of 96 hours. Therefore, the biological results are based on the nominal test item concentration.

 

Endpoint	Nominal concentration
96-h LC50	> 100 mg/L
96-h NOEC	100 mg/L
96-h LOEC	> 100 mg/L

Description of key information

The 96h LC₅₀ is >100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The 96-h NOEC for Penicillin V Potassium is 100 mg/L