DL-α-(aminomethyl)-p-hydroxybenzylic alcohol hydrochloride

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:

31 Jul 2018 to 11 Dec 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

None of the deviations were considered to have impacted the overall integrity of the study or the interpretation of the study results and conclusions.

GLP compliance:

yes

Specific details on test material used for the study:

Identification: Octopamine hydrochloride
Appearance: White to off white powder (determined by Charles River Den Bosch)
Batch: D151-1710037
Test item storage: At room temperature protected from light
Stable under storage conditions until: 26 October 2019 (retest date)

Analytical monitoring:

yes

Details on sampling:

During the limit test, samples for possible analysis were taken from all test concentrations and the control according to the schedule below.
Frequency: 1. At the start of the test and after 48 hours from the freshly prepared solutions.
2. At renewal (t=48h) and the end of the test from the 48-hour old solutions.
Volume: 1.5 mL from the approximate centre of the test vessels
Storage: Samples were stored in a freezer (≤-15°C) until analysis at the analytical laboratory of the Test Facility.

Additionally, reserve samples of 1.5 mL were taken from all test solutions for possible analysis. If not already used, these samples were stored in a freezer (≤-15°C) for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.

Vehicle:

no

Details on test solutions:

The batch of Octopamine hydrochloride tested was a white to off-white powder with a purity of 99.8% and completely soluble in test medium at the concentrations tested. No correction was made for the purity/composition of the test item. Preparation of test solutions started with the highest concentration of 100 mg/L applying a 15-16 minute period of magnetic stirring to accelerate dissolution of the test item in test medium. Lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium. All test solutions were clear and colorless at the end of the preparation procedure. Any residual volumes were discarded.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

Species: Carp (Cyprinus carpio, Teleostei, Cyprinidae) Linnaeus, 1758 Source Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
Mean length1: Range-finding test: 3.2 ± 0.3 cm Final test: 3.5 ± 0.3 cm
Mean weight1: Range-finding test: 0.38 ± 0.11 g Final test: 0.50 ± 0.12 g
Characteristics: F1 from a single parent-pair bred in UV-treated water.
Reason for selection: This system has been selected as an internationally accepted species.
Total fish used: 26

Holding Quarantine/Acclimatisation: At least 12 days after delivery.
Medium: Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, BerkelEnschot, The Netherlands) with the following composition: CaCl2.2H2O 211.5 mg/L MgSO4.7H2O 88.8 mg/L NaHCO3 46.7 mg/L KCl 4.2 mg/L
Measurements Conductivity, pH, nitrate, nitrite and ammonia concentration: once a week. Temperature: continuous. In addition, pH and temperature were measured before transferring the fish to the test system.
Water quality parameters: Were kept within the optimum limits for the respective fish species.
Feeding: Daily with pelleted fish food (Cyprico Crumble Excellent (300500 um), Coppens International bv, Helmond, The Netherlands)
Validity of batch: In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%

1 Ten fish of the batch used for the test, were weighed and measured prior to the start of the test.

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

180 mg CaCO3 per litre

Test temperature:

20-24°C

pH:

6.0-8.5, constant within 1 unit

Dissolved oxygen:

> 60% of air saturation

Salinity:

Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, BerkelEnschot, The Netherlands) with the following composition: CaCl2.2H2O 211.5 mg/L MgSO4.7H2O 88.8 mg/L NaHCO3 46.7 mg/L KCl 4.2 mg/L

Nominal and measured concentrations:

Dose Range Finding Test
Nominal Conc: 0.1, 1, 10 and 100 mg/L

Final Test
Nominal Conc: 100 mg/L
The measured concentrations were at 97-105% of the nominal concentration throughout the test duration.

Details on test conditions:

Range-Finding Test A range-finding test was performed to provide information about the range of concentrations to be used in the final test. Test procedure and conditions were similar to those applied in the final test with the following exceptions:
Three fish per concentration were exposed to a range of nominally 0.10 to 100 mg/L increasing by a factor of 10.
Temperature and pH were only measured in the lowest and the highest test concentration. Dissolved oxygen concentrations were measured in the lowest and the highest test concentration at the start of the test and after 24, 48 and 96 hours of exposure. After 72 hours of exposure, dissolved oxygen concentrations were measured in all test concentrations.
Aeration was introduced after 72 hours of exposure, after dissolved oxygen was observed to be below 5 mg/L in the two highest test concentrations

Test Concentrations: Octopamine hydrochloride 100 mg/L, the regulatory limit concentration. Control Test medium without test item or other additives.

Test Procedure and Conditions
Test duration: 96 hours
Test type: Semi-static, with renewal of test solutions after 48 hours of exposure
Test vessels:20 litres, all-glass, containing 15 litres of test solution
Test medium: Adjusted ISO medium with a hardness of 180 mg CaCO3 per litre and a pH of 7.7 ± 0.3.
Number of fish: 7 fish per concentration and control
Loading: 0.23 g fish/Litre, i.e. 7 fish per 15 litres of test medium
Illumination: 16 hours photoperiod daily
Aeration: The test media were not aerated during the test.
Feeding: No feeding from 24 hours prior to the test and during the total test period.
Introduction of fish: Within five minutes after preparation of the test media from a holding tank with comparable water quality parameters and pH and temperature differences between test and holding tank media of less than 1.0 unit and 1.0°C.
Measurements and Recordings: Mortality and other effects At 2, 24, 48, 72 and 96 hours following the start of exposure. In addition, every afternoon on days 0, 2, 3 and 4, and every morning from day 1 to observe for any dead or severely distressed fish. Dead fish were removed when observed.
Dissolved oxygen content, pH and temperature: Daily in all vessels with surviving fish, beginning at the start of the test (day 0). 5

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (PCP, SIGMA, Art. P2604, Batch MKBW8316V)

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)

Details on results:

Range-Finding test
No mortality or clinical effects were observed at any of the test concentrations during the test period. Based on these results, samples taken from nominal 100 mg/L were analysed. The initial concentration was 103 mg/L. This concentration decreased to 73% of the initial concentration at the end of the test (see also Table 2 of the appended Analytical Report). All test conditions were generally maintained within the limits prescribed by the study plan. After 72 hours of exposure, the dissolved oxygen was below 5 mg/L at the two highest test concentrations. It was consequently decided to introduce aeration for the remaining test period.

Final Test
Measured Test Item Concentrations
Samples taken from the control and the limit concentration were analysed. The measured concentrations were at 97-105% of the nominal concentration throughout the test duration. Based on these results, the nominal exposure concentrations were used to express the effect parameters.

Mortality and Other Effects
No mortality or clinical effects were observed at any of the test concentrations during the test period. The responses recorded in this test allowed for reliable determination of an LC50 and were in agreement with the results of the range-finding test.

Experimental Conditions
All test conditions remained within the ranges prescribed by the study plan (pH: 6.0-8.5, constant within 1 unit; temperature 20-24°C, constant within 2°C; oxygen > 60% of air saturation).

Results with reference substance (positive control):

Incidence of mortality observed in the reference study:
PCP Nominal conc. (mg/L) Initial number Of fish Cumulative number of dead fish recorded at various time points after start of exposure Total Mortality (%)
2½h 24h 48h 72h 96h
0.10 5 0 0 0 0 0 0
0.22 5 0 5 5 5 5 100
0.46 5 0 5 5 5 5 100

Reported statistics and error estimates:

No LC50 could be calculated because the test item proved to be non-toxic (LC50 > maximum concentration).

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the 96h-LC50 was beyond the range tested, i.e. exceeded an analytically confirmed nominal concentration of 100 mg/L.

Executive summary:

The objective of the study was to evaluate Octopamine hydrochloride for its ability to generate acute toxic effects in Cyprinus carpio during an exposure period of 96 hours and, if possible, to determine the LC50 at all observation times. The study procedure described in this report was based on the OECD guideline No. 203, 1992.  The batch of Octopamine hydrochloride tested was a white to off-white powder with a purity of 99.8% and completely soluble in test medium at the concentrations tested. A final test was performed based on the results of a preceding range-finding test. Seven carps per test group were exposed to an untreated control and to nominally 100 mg/L. The total exposure period was 96 hours, test solutions were renewed after 48 hours of exposure and samples for analytical confirmation of exposure concentrations were taken from fresh and old solutions of both refreshment periods.  No mortality or clinical effects were observed at any of the test concentrations during the test period.  Samples taken from the control and the limit concentration were analysed. The measured concentrations were at 97-105% of the nominal concentration throughout the test duration. The study met the acceptability criteria prescribed by the study plan and was considered valid. In conclusion, the 96h-LC50 was beyond the range tested, i.e. exceeded an analytically confirmed nominal concentration of 100 mg/L.  

Description of key information

Seven carps per test group were exposed to an untreated control and to 100 mg/L (nominal) of the test substance. The total exposure period was 96 hours, test solutions were renewed after 48 hours of exposure and samples for analytical confirmation of exposure concentrations were taken from fresh and old solutions of both refreshment periods.  No mortality or clinical effects were observed at any of the test concentrations during the test period.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L