Assays from the EU's Science Committee on Consumer Safety guidance document and from the EPA's conditional approval for HEIQ AG 20S Fred Klaessig, September 25, 2012

OPPTS Algal Toxicity, Tiers I and II Data required: test organisms, including the scientific name, method of verification, strain, and source Control charts of growth in the nontreatment and solvent controls for each toxicity test the number of replicates, the temperature, the lighting, and method of incubation, oscillation rates, and type of apparatus. concentration of the test chemical in the control and in each treatment at the end of the test and the pH of the solutions number of algal cells per milliliter in each treatment and con- trol and the method used to derive these values at the beginning, at 24, 48, and 72 h, and at the end of the test; the percentage of inhibition or stimulation of growth relative to controls 96–h EC50 values, and when sufficient data have been gen- erated, the 24–, 48–, and 72–h EC50s and 95 percent confidence limits, the methods used to derive these values, the data used to define the shape of the concentration-response curve and the goodness-of-fit determination.

OECD 434 Acute Dermal Toxicity – Fixed Dose Procedure tabulation of response data and dose level for each animal (i.e., animals showing signs of toxicity including mortality, nature, severity and duration of effects); individual weight of animals at the day of exposure, in weekly intervals thereafter, and at time of death or euthanasia; date and time of death if prior to scheduled euthanasia; time course of onset of signs of toxicity and whether these were reversible for each animal; time course of onset of signs of toxicity and whether these were reversible for each animal; necropsy findings and histopathological findings for each animal, if available.

OECD 436 Acute Inhalation Toxicity - Acute Toxic Class Method Tabulation of chamber temperature, humidity, and airflow; Tabulation of chamber nominal and actual concentration data; Tabulation of particle size data including analytical sample collection data, particle size distribution, and calculations of the MMAD and σg; Tabulation of response data and concentration level for each animal (i.e. animals showing signs of toxicity including mortality, nature, severity, and duration of effects); Individual body weights of animals collected on study days, date and time of death if prior to scheduled euthanasia; time course of onset of signs of toxicity, and whether these were reversible for each animal; Necropsy findings and histopathological findings for each animal, if available; The GHS category classification and the LC50 cut-off value;

OECD 432 In Vitro3T3 NRU phototoxicity test cell viability obtained at each concentration of the test chemical, expressed in percent viability of mean, concurrent solvent controls; concentration response curves (test chemical concentration vs. relative cell viability) obtained in concurrent +Irr and -Irr experiments; analysis of the concentration-response curves: if possible, computation/calculation of IC50 (+Irr) and IC50 (-Irr); comparison of the two concentration response curves obtained in the presence and in the absence of irradiation, either by calculation of the Photo-Inhibition- Factor (PIF), or by calculation of the Mean-Photo-Effect (MPE); test acceptance criteria; concurrent solvent control ; absolute viability (optical density of Neutral Red extract) of irradiated and non-irradiated cells; historic negative and solvent control data; means and standard deviations; test acceptance criteria; concurrent positive control ; IC50(+Irr) and IC50(-Irr) and PIF/MPE of positive control chemical; historic positive control chemical data: IC50(+Irr) and IC50(-Irr) and PIF/MPE; means and standard deviations.