Insulin Aspart Ethyl Ester

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Read Across please refer to section "justification for type of information"

Justification for type of information:

The aquatic toxicity of Insulin Aspart Ethyl Ester to fish is assumed to follow the same pattern as that of the source substance because of the high degree of structural similarity:
• Insulin Aspart Ethyl Ester and the source substance belong to the same chemical categories: human insulins, polypeptides and proteins
• Insulin Aspart Ethyl Ester and the source substance are composed of polypeptide containing amino acids

Please refer to Environmental Assessment Reports (Section 13)

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 50.1 mg/L

Nominal / measured:

meas. (geom. mean)

Validity criteria fulfilled:

not applicable

Conclusions:

Insulin Aspart Ethyl Ester is predicted to have a toxicity to fish with LC50 of > 50,1 mg/L (geometric mean concentration).

The aquatic toxicity of Insulin Aspart Ethyl Ester is assumed to be similar to the source substance (S4) because of the high degree of structural similarity:
• Insulin Aspart Ethyl Ester and the source substance belong to the same chemical categories: human insulins, polypeptides and proteins
• Insulin Aspart Ethyl Ester and the source substance are composed of polypeptide containing amino acids.

No mortality to fish was observed during the study with Insulin Aspart Precursor (S4) and LC50 is therefore reported as > 50,1 mg/L (geometric mean concentration).

Executive summary:

The aquatic toxicity of Insulin Aspart Ethyl Ester is assumed to be similar to the source substance (S4) because of the high degree of structural similarity:

•       Insulin Aspart Ethyl Ester and the source substance belong to the same chemical categories: human insulins, polypeptides and proteins

•       Insulin Aspart Ethyl Ester and the source substance are composed of polypeptide containing amino acids.

Insulin Aspart Ethyl Ester is predicted to have a toxicity to fish with LC50 of > 50,1 mg/L (geometric mean concentration) based on the results from the study with Insulin Aspart Precursor. No mortality to fish was observed during the study with Insulin Aspart Precursor (S4) and LC50 is therefore reported as > 50,1 mg/L (geometric mean concentration).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017.04.03 till 2017.04.07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

According to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: ISO International Standard 7346-2, 1996. Water Quality – Determination of the acute lethal toxicity of substances to a freshwater fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] Part 2: Semi static method.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline No. 126: Short guidance on the threshold approach for acute fish toxicity. 2010.05.31

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:

Duplicate samples of approx. 5 mL from the control and the test container were collected in 20-mL plastic vials at the initiation of the test and at each sampling time (0h; 48h (before and after water renewal) and 96h). The samples taken were stored at -20 ± 2.0°C and sent frozen to the analytical laboratory.

Vehicle:

no

Details on test solutions:

A stock solution of the test item was prepared by dissolving the test item in Milli-Q water at a concentration of 5 g/L. The stock solution was then dissolved in zebra fish medium to obtain a final test concentration of 125 mg/L.
The reference compound potassium dichromate (K2Cr2O7) was tested at the following concentrations: 0 (control); 50; 100; 200; 300 and 400 mg/L.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The test was performed with zebra fish. The fish were purchased from Credo Fish, Nørresundby, Denmark. The fish arrived at DHI on 2017.02.23

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

none

Post exposure observation period:

not included

Test temperature:

Exposure group: 23.8-24.5°C
Control group: 23.1-24.1°C
The water temperature did not differ by more than ± 1.5°C between test chambers or between successive days at any time during the test

pH:

7.6-8.0

Dissolved oxygen:

88-100%

Salinity:

n.a.

Conductivity:

n.a.

Nominal and measured concentrations:

Nominal: 125 mg/L
Measured concentrations:
T=0 T=48 Old T=48 New T=96
107 mg/L 11 mg/L 105 mg/L 51 mg/L

Details on test conditions:

Limit test
Semi static. Renewal of test media every 48h.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 50.1 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality was observed. Therefore LC50 ( 96h) is reported as > 50,1 mg/L (geometric mean concentration)

Results with reference substance (positive control):

LC50: 200 mg/L

Validity criteria fulfilled:

yes

Conclusions:

No mortality was observed during the study with Insulin Aspart Precursor and LC50 is therefore reported as > 50,1 mg/L (geometric mean concentration)

Executive summary:

A fish acute toxicity study (96h) was conducted with Insulin Aspart Precursor. The study was conducted according to GLP following the OECD Guideline No. 203. The study was conducted as a limit test according to the threshold approach described in the OECD Guideline No. 126.

No mortality was observed during the study. LC50 is reported as > 50.1 mg/L (geometric mean concentration).

Description of key information

The ecotoxicity of Insulin Aspart Ethyl Ester is assumed to be simmilar to the source substance (S4) because of the high degree of structural similarity.

No mortality towards fish was observed during the study with Insulin Aspart Precursor (S4) and LC50 is therefore reported as > 50,1 mg/L (geometric mean concentration).

Insulin Aspart Ethyl Ester is predicted to have a toxicity towards fish of > 50,1 mg/L (geometric mean concentration).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

50.1 mg/L