To Study The Physiological Buffers
In this article, we will discuss to study the physiological buffers 1, Principle 2, Types of buffers 3, Requirements 4, Procedure 5, Calculations 6, Observation table and 7, Precaution for the BSc biotechnology students sem 1
Aim: To study the physiological buffers.
PRINCIPLE:
The buffer solutions are that solution that resists the pH change on the addition of acid or base. Usually, the buffer contains a mixture of a weak acid and its salts such as acetic acid and sodium acetate. For a weak base and its salts such as ammonium hydroxide and ammonium chloride. pH is measured by using pH meter. pH should be inert towards by the molecules. It should be inert towards by the molecules. It should be inert towards by the molecules. It should be stable both enzymatically and hydrolytic, so it should maintain pH over a wider range.
TYPES OF BUFFERS:-
Phosphate buffer:- It is an important buffer activity in the cytoplasm of all cells. It contains sodium dihydrogen phosphate- a proton donor and disodium hydrogen phosphate – a proton acceptor.
Bicarbonate buffer:- It is an important buffer of blood plasma in removing metabolic carbon-dioxide produced. It cuts end in blood plasma and contains carbonic acid – a proton donor and bicarbonate acceptor.
REQUIREMENTS:-
pH meter, distilled water, glass tube, pipettes, sodium phosphate, and disodium phosphate.
Preparation of phosphate buffer:- It contains 2 solutions
Solution A: It contains 0.2M of sodium dihydrogen phosphate that acts as an acid component.
Solution B: It contains 0.2M of disodium hydrogen phosphate that acts as a base component. Now mix ‘x’ml of A and ‘y’ml of B.
PROCEDURE:-
1. Make 200 ml of a solution of 0.2 monobasic sodium dihydrogen phosphate solution A.
2. Make 200ml of the solution of 0.2M of dibasic disodium hydrogen phosphate solution B.
3. For making buffer solution requirements pH mix solution A and solution B in definite proportion as mention in the table.
CALCULATIONS:-
a) 0.2M monobasic NaH2SO42H2O Molecular mass = 156gm
1M of NaH2PO4.H2O =56gm
0.2 Molar of NaH2PO4.2H2O=31.2
Also, 1000ml require=31.2/1000*200 = 6.24gm/200ml
b) 0.2M dibasic Na2HPO42H2O Molecular mass=178gm
1Molar of Na2HPO42H2O=178gm 1Molar of Na2HPO42H2O=178gm/l
0.2 Molar of Na2HPO4.2H2O= 178*0.2 / L= 35.6gm/L 1000ml require Na2HPO4.2H2O=35.6/1000*2000=7.12gm/200ml Preparation of Phosphate buffer= x ml of A + y ml of B= 200
Also, Check Helpful Notes Paper
OBSERVATION TABLE:-
S.no. |
X
(NaH2PO4.H2O) |
Y
(Na2HPO4.2H2O) |
pH |
1 | 93.5ml | 6.5ml | 5.7 |
2 | 77.5ml | 22.5ml | 6.1 |
3 | 39ml | 61ml | 7.0 |
4 | 56.5ml | 43.5ml | 6.1 |
PRECAUTIONS:-
1. Weighing should be proper.
2. pH should be accurately calibrated.
3. Glassware should be clean.
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