Liraglutide precursor

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

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:

2004.04.13

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2012.12.18

GLP compliance:

yes

Specific details on test material used for the study:

The test item is an intermediate. Based on information from the sponsor, the test item is:

The test item is: Liraglutide precursor (NCC 90-0743)

Chemical name: Iso precipitate of liraglutide precursor
CAS No: Not applicable
Molecular formula: C142H216N38O45
Purity: 97-99%
Impurities: Impurities: C and N-terminal truncated derivatives of liraglutide precursor. Percentage of (significant) main impurities:
Typical concentration: 1.2 % w/w
Concentration range: 1.0 – 3.0 % w/w
Form: White powder
Water solubility: Dependent of pH
Log Pow: Not possible to perform
Batch Number: EJ1LK2244
Manufacturing date: 27.11.2016
Stability: In aqueous solution, stable up to 96 hours at 4°C
Origin: Novo Nordisk A/S

Analytical monitoring:

yes

Details on sampling:

Samples were collected from the test solutions at the initiation of the test (0 h), at 24 hours (old and new test solutions) and at the termination of the test (48 h). Samples of approx. 5 mL was collected in 20-mL plastic vials. Samples for chemical analysis were anonymized and only test item and concentration range indicated on the samples. The samples were stored at -20 ± 2.0°C. The samples were sent frozen to the analytical laboratory. The chemical analysis was performed by use of reversed-phase high-performance liquid chromatography (RP-HPLC)

Vehicle:

no

Details on test solutions:

A stock solution of the test item was prepared by dissolving the test item in MQ-water at a concentration of 5 g/L. A test solution for each test concentration was prepared by diluting the stock solution in the test medium

Test organisms (species):

Daphnia magna

Details on test organisms:

A strain of the freshwater crustacean Daphnia magna (Straus) Cladocera, Crustacea, originally isolated from Langedam (Birkerød, Denmark), has been cultured at DHI since 1979

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

renewal of test solution after 24 hours

Post exposure observation period:

not included

Hardness:

272 mg CaCO3/L

Test temperature:

19.2 ± 0.1°C

pH:

7.8 ± 0.2

Dissolved oxygen:

100% of saturation

Nominal and measured concentrations:

The test item will be tested at the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.
Measured concentrations: <0.5; 21.3; 26.4; 35.2; 38.6 mg/L (ND: Not Determined)
The reference compound K2Cr2O7 will be tested at the following concentrations: 0 (control); 0.2; 0.4; 0.7; 1.0; 1.4; 2.0; and 2.8 mg/L

Details on test conditions:

Freshly produced synthetic medium was used in the test. The medium was pre-pared from Milli-Q water according to ISO 6341 Animals <24 hours old were exposed to the test item. At 24 and 48 hours, the number of immobile animals was counted. On this basis, the EC values were calculated. pH and dissolved oxygen concentrations were measured at the initiation of the test, before and after renewal of test solutions at 24 hours and at the termination of the test. Temperature was measured continuously by thermologger.
The immobilization test was performed in glass beakers containing 25 mL of test solution. At the initiation of the test, 5 daphnids <24 hours were added to each container. After 24 hours of exposure, all test solutions were renewed

Reference substance (positive control):

yes

Remarks:

K2Cr2O7

Key result

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

35.2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

no effects were observed during the study

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 38.6 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other:

Remarks:

no effects were observed during the study

Results with reference substance (positive control):

Potassium dichromate (K2Cr2O7)
EC10 (24h) = 0.8 [0.6; 0.9] mg/L
EC50 (24h) = 1.2 [1.1-1.4] mg/L

Validity criteria fulfilled:

yes

Conclusions:

The resulting EC10 (48h) and EC50 (48h) were 35.2 [-; 37.0] and > 38.6 [38.2; 38.6] mg/L respectively (geom. mean conc.)

Executive summary:

A acute toxicity study was conducted according to the OECD guideline no. 201 following GLP.

The study was conducted with the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.

Concentrations were not within 20% of the nomial concentration throughout the study. And endpoints are thus reported as geometric mean concentrations (geo. mean conc.)

The resulting EC10 (48h) and EC50 (48h) were 35.2 [-; 37.0] and > 38.6 [38.2; 38.6] mg/L respectively

Description of key information

A acute toxicity study was conducted according to the OECD guideline no. 201 following GLP.

The study was conducted with the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.

Concentrations were not within 20% of the nomial concentration throughout the study. And endpoints are thus reported as geometric mean concentrations (geo. mean conc.)

The resulting EC10 (48h) and EC50 (48h) were 35.2 [-; 37.0] and > 38.6 [38.2; 38.6] mg/L respectively

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

38.6 mg/L