General Exposure Conditions Sample Clauses

General Exposure Conditions. The disc cuts were performed the day before the exposure to RTLF and were kept at room temperature and in a dark place. On the day of the exposure 450μl Chelex-treated water was added, and samples were briefly vortexed. It was checked that all disc cuts were fully immersed in the solution. The PM-filter solutions were then pre-incubated at 37°C (Stuart Scientific Incubator S160) for 10 min prior to a short vortex mixing, the addition of 50μl of the 2mM RTLF stock solution (containing 2mM AA, GSH and UA; see RTLF section) to each incubation tube and a further vortex mix. This process resulted in a final starting concentration of 200µM antioxidants. The resulting reaction mixtures were incubated for up to 2 hours at 37°C with gentle mixing throughout (Vortex Genie-2, shaking mode). Controls were also incubated (see relevant section). For each time point samples were centrifuged at 13,000rpm for 1 hour at 4°C and the supernatant analysed for remaining antioxidant (HPLC and Glutathione analysis sections).