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Gluskin 6 Abstract: This experiment attempts to answer the question of whether an increase in a succinate concentration (a component of the Krebs cycle) will lead to an increased rate of cellular respiration within a cell. We measured the amount of electrons given off by the succinate to fumerate redox reaction by using DPIP. DPIP is an electron acceptor that takes the place of FAD by accepting the electrons and turning from its oxidized blue state to its reduced clear state. We had three tubes with varying concentrations of succinate and measured the transmittance of each over a half hour period to determine whether more succinate led to more DPIP being reduced. The results showed that the tube with no succinate (no …show more content…

This ensured that our experimental samples were only showing readings for the light absorbed by the DPIP and not by the mitochondrial suspension as well. Tube 1 consisted of 4.4 ml buffer, 0.3 ml DPIP, 0.3 ml mitochondrial suspension, and 0 ml of succinate. This served as a control because there was no succinate to give off electrons and reduce the DPIP. Tube 2 consisted of 4.3 ml buffer, 0.3 ml DPIP, , 0.3 ml mitochondrial suspension and 0.1 ml succinate. This was the trial with a low concentration of substrate. Tube 3 had 4.2 ml buffer, 0.3 ml DPIP, 0.3 ml mitochondrial suspension and 0.2 ml succinate. This final tube was used to notice the effect of increased substrate on the amount of electrons given off, or the amount of DPIP reduced. We also made sure to add succinate to each tube last, so that the succinate to fumerate reaction would not occur before the DPIP was there to accept the electrons. Immediately after creating each cuvette, we covered it in parafilm to minimize contamination and placed it directly in the spectrophotometer to get an accurate reading. We recorded the absorbance of cuvettes 1, 2, and 3 every 5 minutes for the next thirty minutes and recorded the absorbances in a table. We also made sure to properly mix the contents of the cuvettes thoroughly before getting a reading in order to best distribute the DPIP throughout the solution. Results: Transmittance of

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