Insulin Human Methyl Ester

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:

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: Hi-OMe (Human Insulin Methylester)
Chemical name: Zinc crystals of HI-OMe
CAS No: Not applicable
Molecular formula: C258H385N65O77S6
Purity: 95-98%
Impurities: Derivatives of HI-OMe, desB30-Insulin, Insulin Human
Percentage of (significant) main impurities:
Typical Concentration: <3.0 % w/w
Concentration Range: 2.0 – 5.0 % w/w
Form: White powder
Water solubility: Dependent of pH
Log Pow: Not possible to perform

Batch Number: EM0Z31503
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:

yes

Remarks:

HCl

Details on test solutions:

A stock solution of the test item was prepared by dissolving the test item in 0.01 M HCl 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 test solution was filtered through a 0.45 μm filter to avoid precipitation.
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: 23.1-24.1°C
Control group: 23.5-24.2 °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:

7.3-8.0

Dissolved oxygen:

73-100%

Salinity:

n.a.

Conductivity:

n.a.

Nominal and measured concentrations:

Nominal: 125 mg/L
Measured:
T=0 T=48 Old T=48 New T=96
18,5 mg/L 0,9 mg/L 13,9 mg/L 0,7 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.:

> 3.6 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 values ( 96h) are reported as > 3.6 mg/L

Results with reference substance (positive control):

LC50: 200 mg/L (nominal concentrations)

Validity criteria fulfilled:

yes

Conclusions:

No mortality was observed during the study with Insulin Human Methyl Ester and LC50 is therefore reported as > 3.6 mg/L (geometric mean concentration)

Executive summary:

A fish acute toxicity study (96h) was conducted with Insulin Human Methyl Ester. 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 > 3.6 mg/L (geometric mean concentration).

Description of key information

A fish acute toxicity study (96h) was conducted with Insulin Human Methyl Ester. 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 > 3.6 mg/L (geometric mean concentration).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

3.6 mg/L