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First instar Chironomus riparius larvae (in house breeding at BT) are exposed to a range of concentrations of the test substance in the test medium (M4 medium) for a period of 48 hours. The exposure is carried out under controlled conditions of temperature and light cycle. The immobilization is recorded at 24 and 48 hours and compared with the control.
A toxic reference substance is tested twice a year to check the sensitivity of the organisms.
Range finding test includes at least 5 concentrations of the test substance and 1 untreated control (2 or 4 replicates/concentration or control) with 5 larves/replicate
Limit test is carried out with 1 concentration (100 mg/L or at solubility limit) of the test substance and 1 untreated control (4 replicates/concentration or control) with 5 larves/replicate
Dose-response test includes at least 5 concentrations of the test substance and
1 untreated control (4 replicates/concentration or control) with 5 larves/replicate
Analytical checks, to verify the exposure concentrations in the medium, are performed at time 0 and at time 48 hours on control and on each test substance sample.
The analysis are performed using a method validated according to SANCO/825/00 rev. 8.1 and SANCO/3029/99 rev. 4 guidance documents.
Immobilization: EC50 at 48 hours as the main endpoint and as other endpoints: EC10, EC20, LOEC/NOEC.
Study includes GLP management and reporting.
REFERENCES AND GUIDELINES
OECD Guideline fo Testing of Chemicals, No. 235 (28th July 2011) - Chironomus sp., acute immobilization test.
SANCO/825/00 rev. 8.1 (16/11/2010) - Guidance document on pesticide residue analytical methods.
SANCO/3029/99 rev. 4 (11/07/2000) - Residues: guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (Part A, section 4) and Annex III (Part A, section 5) of Directive 91/414.