1. Preparation of the buffer solutions, titration of acetic acid and its buffering capacity

2. Objectives To study the nature of the buffers To investigate the buffering capacity of acetic acid by doing the titration curve.

3. Buffers Buffers are the solutions which resist changes in pH when small amounts of acid or alkali is added to them. A buffer is a pair of weak acid and its salt with a strong base. Buffers are of main importance in regulating the pH of the body fluids and tissues Many biochemical reactions including those catalyzed by enzymes require pH control which is provided by buffers

4. Buffers Mammalian tissues in the resting state have a pH of about 7.4 In order to maintain the required pH in an invitro biochemical experiment a buffer is always used The pH of a buffer is given by Handerson- Hasselbalch equation pH= pKa + log [A-] [HA]

5. Buffers pKa= -log Ka Ka is the dissociation constant of the acid [A-] is the concentration of the base [HA] is the concentration of the acid

6. Buffers A buffer consisting of a mixture of the weak acid(CH3COOH) and its salt (CH3COONa+) Will undergo the following changes on the addition of acid or base: If the base is added (OH) to the solution it will be buffered by the following reaction with acetic acid: CH3COOH + OH  CH3COO- + H2O So the pH will not change significantly

7. Buffers If acid (H+) is added, it will be buffered by another reaction, this time using the salt (CH3COO-) CH3COO- + H+  CH3COOH The pH will not alter significantly because the CH3COOH formed is a weak acid Addition of more base increases A- and decrases (HA) and this doesn’t alter the pH much until [A-]>>>>[HA]

8. Buffers The buffering power is greatest when pH=pKa i.e. when the acid and the salt are at the same concentration

9. Method You are provided with 0.1M sodium acetate and 0.1M acetic acid solution Prepare 45ml of the buffer solution by mixing the two standard solutions Use the following info to calculate the composition of the buffer: pH of the buffer 4.86 pKa of acetic acid at 25C 4.76

10. Buffer Preparation of buffer measure 24ml of 0.1M sodiumacetate measure 20.25ml of 0.1M acetic acid mix them complete the volume to 45 ml with dis water measure the pH( calibrate the pH meter before measuring) Checking the buffering capacity with HCL add 1ml HCL to the buffer measure the pH and record it in the table in the result section

11. Titration Titration curves are produced by monitoring the pH of given volume of a sample solution after successive addition of acid or alkali The curves are usually plots of pH against the volume of titrant added or more correctly against the number of equivalents added per mole of the sample

12. Titration Determination of pKa values: pKa values can be obtained from the titration data by the following methods: 1.The pH at the point of inflection is the pKa value and this may be read directly 2.By definition the pKa value is equal to the pH at which the acid is half titrated. The pKa can therefore be obtained from the knowledge of the end point of the titration.

13. Titration