Insulin Aspart 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

Remarks:

According to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004.04.13

Deviations:

no

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: Insulin Aspart (IA) Precursor (NN2000 precursor)

Chemical name: Iso precipitate of IA precursor
CAS No: Not applicable
Molecular formula: C302H446N76O91S7
Purity: 90-99%
Impurities: Derivatives of IA precursor
Percentage of (significant) main impurities:
O-glycosylated derivatives, desamido derivatives and ox-idated derivatives
Typical concentration: 4.0 % w/w
Concentration Range: 1.0 – 10.0 % w/w
Form: White powder
Water solubility: Dependent of pH
Log Pow: Not possible to perform
Batch Number: K2, FHIAZG135 (undissolved substance)
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:

In order to verify the test concentrations, 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 were collected in 20-mL plastic vials. Samples for chemical analysis were anonymized and only test item and concentration range was indicated on the samples. The samples were then 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:

The test item was tested at the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.
The reference compound K2Cr2O7 was tested at the following concentrations: 0 (control); 0.2; 0.4; 0.7; 1.0; 1.4; 2.0; and 2.8 mg/L

Test organisms (species):

Daphnia magna

Details on test organisms:

Juvenile organisms < 24 h old at the beginning of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

The test was run as a semi-static test with renewal of all test solutions after 24 hours.

Post exposure observation period:

not included

Hardness:

250 ± 25 mg/L expressed as CaCO3

Test temperature:

19.2 ± 0.1°C

pH:

7.9 ± 0.1

Dissolved oxygen:

96-100% saturation

Salinity:

n.a.

Conductivity:

n.a.

Nominal and measured concentrations:

The test item was tested at the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.
Meassured concentration: Since no effects were observed only samples from the highest concentration applied were analysed.
T= 0h T= 24h (old) T= 24h (new) T=48h Geom. Mean
257.1 259.5 255.9 246.8 254.8 mg/L

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 300 mg/L

Nominal / measured:

nominal

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:

LC10

Effect conc.:

300 mg/L

Nominal / measured:

nominal

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:

EC10

Effect conc.:

> 254.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 254.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No effects were observed. Test concentrations were within within +/- 20 % of the nominal concentration throughout the test

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:

No mortality was observed during the study, thus an EC10 and an EC50 > 300 mg/L (nominal concentration) (> 254.8 mg/L (geom. mean concentrations)) was determined respectively.

Executive summary:

A Daphnia sp. acute immobilization test (according to OECD Guideline no. 202) was conducted applying daphnids <24h old. The study was conducted according to GLP

The test item was tested at the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.

Organisms were exposed to the test item for 48h and since no mortality was observed during the study an EC10 and EC50 > 300 mg/L (nominal concentration) (> 254.8 mg/L (geom. mean concentrations)).

Description of key information

A Daphnia sp. acute immobilization test (according to OECD Guideline no. 202) was conducted applying daphnids <24h old. The study was conducted according to GLP

The test item was tested at the following nominal concentrations: 0 (control); 20; 40; 75; 150 and 300 mg/L.

Organisms were exposed to the test item for 48h and since no mortality was observed during the study an EC10 and EC50 > 300 mg/L (nominal concentration) (> 254.8 mg/L (geom. mean concentrations)).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

254.8 mg/L