Practical Exercise 3 – Buffers and Titration of An Amino Acid
Name: Stephanie Tayas Anak Bumphray
Id Number: 100084790
Date Performed: 11 September 2017
Practical Group: Monday 1:30pm – 5:30pm
Introduction:
A buffer solution is which resists large changes in pH when the small amounts of an acid or a base are added to it. The buffer can be calculated based on the equation of ionization pair of acid and conjugate base. Any weak acid can be represented as “HA” and "A-" as the conjugate base. This equation is based known as Henderson-Hasselbalch equation which is used to calculate pH of a solution. Figure 1: Henderson-Hasselbalch equation
Buffer solution can be divided into two which are acidic buffer solutions and alkaline
…show more content…
In exercise 1, first, the ratio of [HPO42-] to [H2PO4-] required to produce buffer solutions was calculated with the following pH values (a) pH = 5.9, (b) pH = 7.9. Then, the solutions of 0.1 M H2PO4- and 0.1 M HPO42- was used, these buffer solutions were mixed appropriate volumes to give a total volume of 25 mL of each. The calibrated pH meter was used, the pH of each buffer solution was measured and recorded and the pH of 25mL of distilled water was compared. Next, 2.0 mL of 0.1 NaOH was added to each of the 25mL buffer samples and to the distilled water and each tube was mixed well. The pH of each solution was recorded after the addition of the alkali. Results was tabulated include a column for ∆ …show more content…
= 22.7mL
Volume of base required = 25mL – 22.7mL = 2.3mL
(b) pH = 6.9 pH = pka + log10 [HPO42-] 6.9 = 6.9 + log10 [HPO42-] [H2PO4-] [H2PO4-] log10 [HPO42-] = 0 [H2PO4-] = 1 1
Volume of acid required = 25mL (1+1) = 12.7mL
Volume of base required = 25mL – 12.7mL = 12.5mL
(c) pH = 7.9 pH = pka + log10 [HPO42-] 7.9 = 6.9 + log10 [HPO42-] [H2PO4-] [H2PO4-] log10 [HPO42-] =
In this lab, the purpose was to determine the stability of a substance after adding an acid or a base. The results claim that liver and buffer are the most resistance to change in pH. Looking at figure 3, buffer and liver both maintain a stable pH even with the addition of an acid or base. However, potato and water have less buffer in them since their pHs did change. In figure 3, the potato acid’s pH level decreased by two, and the potato base’s pH level increased by two. The level of pH of a water acid decreased by 4, while the water base’s pH increased by 5. These results all tie to the fact that buffer is a substance that maintains a stable pH; the presence of buffer in organisms help maintain homeostasis by binding or releasing hydrogen
1. To titrate a hydrochloric acid solution of “unknown” concentration with standardized 0.5M sodium hydroxide.
After added, pick up the beaker and swirl it around lightly for a short period of time.
pH was recorded every time 1.00 mL of NaOH was added to beaker. When the amount of NaOH added to the beaker was about 5.00 mL away from the expected end point, NaOH was added very slowly. Approximately 0.20 mL of NaOH was added until the pH made a jump. The pH was recorded until it reached ~12. This was repeated two more times. The pKa of each trial are determined using the graphs made on excel.
A buffer is a solution that resists changes in pH when H+, OH-, or H20 is added. By using standard lab equipment, a lab pro diagnostic tool, and acidic and basic solutions, the pH can be found. By recording the pH while adding a base or an acid gradually to a buffer solution you can find the capacity of each buffer to resist drastic changes in pH. The best buffers will keep a solution from becoming either too acidic or basic with the addition of a strong base or acid.
For the peptide Ala-Arg-Lys-Ala-Asn-Ser-Ala-Ser, what would be the expected charges at pH 1, 7, and 13?
To improve the results from the experiment buffer solutions that were not whole pHs could have been used e.g. pH 4.5, 5.5 etc. This would have provided more reliable results as a wider range of results would have been produced. Using pHs with decimals would also help to more accurately determine the optimum pH as the optimum may have been above or below the pH stated in the hypothesis; 8. In this experiment however the optimum is taken at 8 because the graph does not rise again.
To make the buffer solution you need 0.2 mol dm-3 of Na2HPO4 and 0.1mol dm-3 of citric acid this will give 100cm3 of buffer. Here is how to get the different pH in the buffer solutions:
ii. The second part of the titration series involves titration of NaOH with Hydrochloric acid (HCL). Again, three reps of titration and a blank titration have to be completed. A volumetric pipet is used to measure 10.00mL of HCL into three labeled conical flasks. Then the flasks are filled with deionized water until about the 50mL mark. A buret is
Within an acid-base titration the titration curve resembles the strengths of the corresponding acids and bases. A strong acid will correspond with a weak conjugate base, and a weak conjugate acid will correspond with a strong base. This is based on the Bronsted-Lowry model. The weak acid will donate protons to the hydroxide ion. Weak acids will have a low Ka value, the Ka value is the tendency of the acid to dissociate:
The pH of a solution is the measure of the concentration of charged Hydrogen ions in that given solution. A solution with a pH lower than seven is considered to be acidic. A solution with a higher pH is a base. It is very important for organisms to maintain a stable pH. Biological molecules such as proteins function only at a certain pH level and any changes in pH can result in them not functioning properly. To maintain these constant pH levels, buffer solutions are used. A buffer solution can resist change to small additions of acids or base’s. A good buffer will have components that act like a base, and components that act like an acid.
For this experiment, titrations on a weak acid, acetic acid, and a buffer were performed. Acetic acid was titrated with NaOH in order to observe the half-equivalence point as well as the equivalence point. Then, the buffer and the buffered acetic acid solution prepared faced additional titration with NaOH and HCl to evaluate the differing buffering effects following the addition of a strong acid and strong base. Finally, the buffer’s buffering capacity was calculated. If the experiment were to be repeated, it would be interesting to observe the buffering effects following a titration between a weak base and a buffer instead with greater concentrations. The change in the concentration following the preparation of buffer with a weak base and its conjugate acid would pose for an interesting experiment to observe an increase in the buffering capacity.
The titration curve of the unknown exhibited many characteristics, such as equivalence points, pKa of ionizable groups, isoelectric point, and buffer regions, that are particularly distinct to lysine. For unclear reasons, the pH during the titration did not reach the pH for pure 0.2 M NaOH nor 0.2 M HCl and normal equivalence points expected at two extreme ends of the titration curves for all amino acids were not observed. The titration of a phosphate buffer showed that the buffer capacity is directly proportional to the molarity of the buffer. However, our results showed that although the initial pH of the phosphate buffer was less than the pKa value, the measured buffer capacity was higher towards acid than base. The accuracy of the pH meter and calibration process was questioned under assumptions that the pH of the prepared phosphate buffer was actually above pKa.
2. Following solutions are added to the tubes and the pH of each tube is determined:
In this lab a acid-base indicator phenolphthalein was used to determine endpoint of a reaction HCl(aq) and KOH(aq). At the end point all of the HCl(aq) would have reacted with KOH(aq), and the pH becomes 7. The phenolphthalein would changed colours from colourless to pink indication when enough KOH(aq) was added. The purpose of numerous trials was to use the average volume of the 3 trials with similar measurements.