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The mobile phases con¬sisting of methanol and water were not efficient enough to clearly separate all fractions. Using a mobile phase with acetonitrile as a non-polar solvent resulted in a better sep¬aration of different peaks, where the actual volume fraction of acetonitrile depended on the analyzed samples. We first analyzed LFP obtained after in vitro incubation of mito¬chondria with MDA at 25 °C. Comparing the chromato¬grams obtained with different volume fractions of acetonitrile used, it was found that mobile phase made of 60% acetonitrile, v/v, gave the best separation of individual peaks. Using the same procedure we found that the best separation of LFP from mitochondria incubated with MDA at 50 °C was with 40% acetonitrile, v/v. |
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