Answers to Postlab- Questions A-1.) My measured pH values for 0.01N NaOH and 0.

01N KOH are different because the K+ ions and Na+ ions have different atomic sizes. Since K+ ions are larger than Na+ ions because it has more protons, its atomic diameter is bigger. This means that the bond between K+ ions and the hydroxide is longer because it has less ability to pull the electrons on the oxygen towards the nucleus. Since the bond is longer, it is weaker, and thus K+ ions are supposed to ionize more than Na+ ions. Therefore, because of the ways KOH and NaOH dissociate are different, their pHs also differ, since the pH is the measure of [H+], or in other words, 14 pOH. Also, NaOH can absorb CO2 from the atmosphere, which will affect the pH when the CO2 dissolves in solution. A-2.) They were both different from the theoretical value, which is pH = 12, because there could have been an error with the pH meter, or maybe the solution of KOH and NaOH got mixed with water during transfer, or when we calibrated the pH meter, it was not actually pH4. Since the calibrated pH meter will calibrate exactly at pH4 and pH7 for solutions that are marked pH4 and pH7, if those solutions are not pH4 and pH7, even if the pH value of the NaOH and KOH exactly 12, it will not measure it as 12. A-3.) KOH was used instead of NaOH as a titrant because NaOH can quickly absorb CO2 from the atmosphere, which can form NaCO3. This will throw off the pH unless you are able to keep NaOH in a closed system.

C-1.) Since trypsin is in HCl, the extra H+ from the HCl can increase the number of H+ ions, making it seem like the number of peptide bonds is more than it is. Therefore, it may appear as if there is an increased number of residues of lysine and arginine. Results: The pH curve (Graph 1) showed four areas of potential flat lines, also the isoelectric points (pI). This can also be seen on Graph 2, which is water adjusted. The areas where buffer capacity is high around pH= 1.25, pH = 6.00, pH=10.25, and pH=12.25. The graph for Time vs. mmols of KOH show an acute increase in KOH addition until around 10 minutes until it starts to become more stable. The endpoint is at 35 minutes when the amount of KOH added was 0.219 mmols. Discussion: The theoretical value for B-2 was very close to what we obtained experimentally. The error was 0.5%, which is shown in the calculations in question B-2. Our standard curve was not as good as it could have been because we did not take the time to take more points between pH6~12. Therefore, even though there is some indication of a bump in the middle, it is hard to see. However, to estimate, I still used the tiny bump that appears near 9.18 pH as the third buffering capacity zone. In order to calculate the second capacity zone at around ~pH6, I averaged two data points of (5.75mmol, 5.74 pH) and (8.25 mmol and 6.11pH) to get 5.92pH.

B-1.) Calculations shown below

B-2.) The theoretical buffer capacity for the histidine buffer at pH 6.0

B-3.)
pH1 pH6.2 pH9.8 pH11

B4.) I am not completely sure, but my thought is that this value could be experimentally confirmed if one did a titration of pure histidine buffer without putting in any water. If they do it this way, then the curves should look the same because the curve calculated on Graph 2 is adjusted for water error. B-5.) The experiments that would help confirm the identity of the groups responsible are experiments that separate according to charges. Therefore, experiments such as cation exchange and anion exchange would be able to separate the positively charged histidine molecules and the negatively charged histidine molecules, respectively.