6-aminopenicillanic acid

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:

2009-11-12 to 2009-11-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: A GLP study conducted to current accepted testing guidelines.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP standards (Schedule 1, Good Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by SI 2004/0994)).

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not reported

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: Water samples were taken at 0 24 and 96 hours from the test and control vessels. Samples were also taken in duplicate at 24, 48 and 72 hours and stored for further analysis where nececessary.
- Sample storage conditions before analysis: The additional samples were stored at -20 °C

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: An amount of test material (2000 mg) was dissolved in dechlorinated tap water and the volume adjusted to 1 litre to give a 2000 mg/L stock solution. This stock solution was dispersed in 20 litres (final volume) of dechlorinated tap water, and stirred using a flat bladed mixer for approximately 1 minute, to give the required test concentration of 100 mg/L.
- Controls: The control group were maintained in identical conditions to the test groups, but where not exposed to test material.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Brow Well Fisheries Limited, Hebden, (near Skipton), Yorkshire, UK
- Age at study initiation: Stated as juvenile fish
- Length at study initiation : mean standard length of 4.3 cm (sd = 0.4) at the end of the definitive test
- Weight at study initiation: mean weight of 1.10 g (sd = 0.28) at the end of the definitive test.
- Feeding during test: No feeding during the test
- Food type: The stock fish were fed commercial trout pellets which was discontinued 24 hours prior to the start of the definitive test.

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: Test conditions
- Type of food: commercial trout pellets
- Health during acclimation : No mortalities were observed 7 days prior to study initiation

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Hardness:

Approximately 140 mg/L as CaCO3

Test temperature:

Please refer to table 2 Physico-Chemical Values of Test Conditions

pH:

Please refer to table 2 Physico-Chemical Values of Test Conditions

Dissolved oxygen:

Please refer to table 2 Physico-Chemical Values of Test Conditions

Salinity:

Not reported

Nominal and measured concentrations:

In the range finding study, due to a dosing error, a nominal concentration of 50 mg/L was employed.
In the definitive test, a nominal concentration of 100 mg/L was used nominal concentrations were found to be equal to measured concentrations.

Details on test conditions:

TEST SYSTEM
- Test vessel: 20 litre glass exposure vessels
- Type: semi-static system, covered to prevent evaporation
- Aeration: the test vessels were aerated via narrow bore glass tubes.
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 7
- No. of vessels per concentration : 2 replicates of 100 mg/L. In the range finding test only one group at 50 mg/L was used.
- No. of vessels per control: One control group was employed in each of the range finding and definitive tests.
- Biomass loading rate: Based on the mean weight value this gave a loading rate of 0.39 g bodyweight/litre.

TEST MEDIUM / WATER PARAMETERS
Typical water characteristics are recorded in table 3.

OTHER TEST CONDITIONS
- Photoperiod: photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours.

EFFECT PARAMETERS MEASURED : The water temperature, pH and dissolved oxygen concentrations were recorded daily throughout the test. The measurements at 0 hours, and after each test media renewal at 24, 48 and 72 hours, represent those of the freshly prepared test preparations while the measurements taken prior to each test media renewal, and on termination of the test after 96 hours, represent those of the used or 24-Hour old test preparations. The pH and the dissolved oxygen concentration were measured using a WTW pH/Oxi 340I pH and dissolved oxygen meter and the temperature was measured using a Hanna Instruments HI93510 digital thermometer .Any mortalities and sub-lethal effects of exposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start of exposure. The criteria of death were taken to be the absence of both respiratory movement and response to physical stimulation.

TEST CONCENTRATIONS
- Range finding study: 50 mg/L in the range finding study
- Test concentrations: 100 mg/L (limit test) in the definitive study. Analysis of the test preparations at 0, 24 and 96 hours showed measured test concentrations to be near nominal and so the results are based on nominal test concentrations only.
- Results used to determine the conditions for the definitive study: The results of the range-finding test showed no mortalities at the test concentration of 50 mg/l. Together with the results of the range-finding test conducted for the Acute Toxicity to Daphnia magna study on this test material (Harlan Laboratories Project Number 1127/1849), it was considered appropriate to perform the definitive test as a "Limit test" at a concentration of 100 mg/l to confirm that at the maximum concentration given in the OECD/EEC Test Guidelines

Reference substance (positive control):

no

Remarks:

Not a requirement of the guideline

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

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: No sub-lethal abnormalities were noted.
- Mortality of control: No mortality observed in the controls in both the range finder and definitive test.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Throughout the test the test preparations were observed to be clear colourless solutions. A single dissolved oxygen concentration (Control, 48-Hour, old media) was measured to be in excess of 100% ASV. This was considered to be due to microscopic air bubbles supersaturating the test media and was considered to have no adverse effect on the test as the control fish showed no adverse effects. While there were no treatment related differences for oxygen concentration, the pH of the test material preparations was observed to be lower than that of the control at each media renewal. This was considered to be due to the acidic nature of the test material. The light stability result (76%) is slightly below the acceptance criteria of 80 to 120%. However, this result is not considered to be significantly different from the results under other stability conditions, and therefore does not suggest significant degradation by photolysis. The results do indicate that the test material is slightly unstable in the test medium over 24 hours. This does not match the results obtained at 100 mg/l in the Acute Toxicity to Daphnia magna or Algal Growth Inhibition Test (Harlan Project Numbers 1127/1849 and 1127/1850 respectively), which showed the test material to be stable. The Algal Growth Inhibition Test did however show the test material to be unstable at lower concentrations. The differences in stability results are not fully understood, but are believed to be most likely attributable to differences in test media. The unsonicated stability vessel showed no evidence of insolubility or adherence to glass.

Results with reference substance (positive control):

Not reported

Reported statistics and error estimates:

Not reported

Sublethal observations / clinical signs:

Tabulated results are presented in the attached pdf Appendix I Tabulated Data for Goodband TJ and Mullee DM 2010

Table 9: Summary of Results

Time (h)	LC50(mg/L)
3	>100
6	>100
24	>100
48	>100
72	>100
96	>100

Validity criteria fulfilled:

yes

Remarks:

All of the validity/quality criteria set out in OECD 203 and EU C.1 were met within the study.

Conclusions:

The acute toxicity of the test material to the freshwater fish rainbow trout (oncorhynchus mykiss) was investigates and gave a 96 hour LC50 of greater than 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.

Description of key information

The short term toxicity to fish of 6-Aminopenicillanic acid was determined by OECD Guideline 203 (Fish, Acute Toxicity Test).

Additional information

The acute toxicity of the test material to the freshwater fish rainbow trout (oncorhynchus mykiss) was investigates and gave a 96 hour LC50 of greater than 100 mg/L. Correspondingly the No Observed Effect Concentration was 100 mg/L.