Liraglutide precursor

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

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 201 (Alga, Growth Inhibition Test)

Version / remarks:

2004.04.13

Qualifier:

according to guideline

Guideline:

ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)

Version / remarks:

2012.02.15

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:

The actual exposure concentrations of the test item was determined by the sponsor’s analytical laboratory. Duplicate samples of approx. 5 mL from the specially prepared test flasks were collected in 20-mL plastic vials at the initiation of the test and at each sampling time (24h; 48h and 72h). Samples for chemical analysis were anonymized and only test item and concentration range indicated on the samples. The samples taken were stored at -20 ± 2.0°C and 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 is prepared by dissolving the test item in MQ-water at a concentration of 5 g/L. A test solution for each test concentration will be prepared by diluting the stock solution in algal test medium.

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Details on test organisms:

Pseudokirchneriella subcapitata (NIVA-CHL 1) is cultured at DHI

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Post exposure observation period:

Not included

Test temperature:

23 ± 2.0°C in a climate room under uniform continuous illumination from a panel of fluorescent light with an intensity of 60-120 µmol/m2 × s

pH:

8.1 ± 0.2

Nominal and measured concentrations:

The test item will was tested at the following nominal concentrations: 0 (control); 5; 10; 20; 40; 75; 150 and 300 mg/L
measured cocnentration: 1.3; 4.1; 11.5; 20.5; 15.4; 24.0 35.4 mg/L

Reference substance (positive control):

yes

Remarks:

The reference compound potassium dichromate (K2Cr2O7) will be tested in the following concentrations: 0 (control); 0.1; 0.2; 0.4; 0.7; 1.0; 1.4 and 2.0 mg/L.

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

< 1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

< 1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EC20

Effect conc.:

< 1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

28.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

Values based on nominal concentrations

NOEC: <5 mg/L
LOEC: ≤5 mg/L

EC values and limits of the 95% confidence interval
EC10: <5 mg/L
EC20: <5 mg/L
EC50: 82 mg/L (39 - >300 mg/L)

Results with reference substance (positive control):

Potassium dichromate (K2Cr2O7)
NOEC = 0.1 mg/L
LOEC = 0.2 mg/L

Validity criteria fulfilled:

yes

Conclusions:

Based on the endpoint growth rate a LOEC and a NOEC of ≤1.3 mg/L and <1.3 mg/L respectively and an EC10, EC20 and EC50 (72h) of <1.3, <1.3 and 28.5 mg/L (geom. mean concentration) were determined during the study.

Executive summary:

An acute toxicity study with algae (Pseudokirchneriella subcapitata) was conducted according to the OECD guideline No. 201 following GLP.

The following nominal concentrations were tested: 0 (control); 5; 10; 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.)

Based on the endpoint growth rate a LOEC and a NOEC of ≤1.3 mg/L and <1.3mg/L respectively and an EC10, EC20 and EC50 (72h) of <1.3, <1.3 and 28.5 mg/L (geom. mean concentration) were determined during the study.

Description of key information

An acute toxicity study with algae (Pseudokirchneriella subcapitata)was conducted according to the OECD guideline No. 201 following GLP.

The following nominal concentrations were tested: 0 (control); 5; 10; 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.)

Based on the endpoint growth rate a LOEC and a NOEC of 1.3 mg/L and <1.3mg/L respectively and an EC10, EC20 and EC50 (72h) of <1.3, <1.3 and 28.5 mg/L (geom. mean concentration) were determined during the study.

Key value for chemical safety assessment

EC50 for freshwater algae:

28.5 mg/L