ADANA ALPARSLAN TÜRKEŞ SCIENCE AND TECHNOLOGY UNIVERSITY

DEPARTMENT OF BIOENGINEERING

MOLECULAR BIOLOGY LABORATORY

EXPERIMENT 1. Buffers & Phosphate Buffer Preparation

Date of Experiment : 15.10.2021

Date of Submission : 22.10.2021
Student Name:Betül Oğuzbir
Instructors : Burcu Eren,Okan Zenger
Student Number:190102004
INTRODUCTION
A buffer is a solution containing either a weak acid and its salt or a weak base and its salt,
which is resistant to changes in pH. In other words, a buffer is an aqueous solution of either a
weak acid and its conjugate base or a weak base and its conjugate acid. A buffer may also be
called a pH buffer, hydrogen ion buffer, or buffer solution.
Buffers are used to maintain a stable pH in a solution, as they can neutralize small quantities
of additional acid of base. For a given buffer solution, there is a working pH range and a set
amount of acid or base that can be neutralized before the pH will change. The amount of acid
or base that can be added to a buffer before changing its pH is called its buffer capacity. 
The Henderson-Hasselbalch equation may be used to gauge the approximate pH of a
buffer. In order to use the equation, the initial concentration or stoichiometric concentration is
entered instead of the equilibrium concentration.

The general form of a buffer chemical reaction is:

HA ⇌ H+ + A−

AIM
To prepare the buffer at required pH.

EXPERIMENTAL

MATERIALS &METHOD
-Monosodium phosphate
- Disodium phosphate
- Water
-Phosphoric acid to make the pH more acidic or sodium hydroxide to make the
-pH more alkaline
- pH meter13
- Glassware
- Hot plate with stirring bar

METHOD

1-)The solution was prepared by mixing 0.477 moles of monosodium phosphate and

2-)0.523 moles of disodium phosphate prepared in a little less than one liter of water.

3-)The acid solution weighed 28.61 grams.

4)-The base solution measured 37.13 grams.

5-)The pH of the mixture was first measured at 6.60.

6-)Base solution was added to make the pH 7.

7-)Volume completed to 500 ml.( First we made it in 400 ml ) .

RESULTS

|Acid| |Base|

0.477 moles/L(molarity) 0.523 moles/L(molarity)

28.61 grams(weighed) 37.13 grams(weighed)

DISCUSSION

If acid is added to a solution, the pH tends to be lower than its original pH, and if a base

solution is added, the pH change will be greater than the initial value. They are very different

from each other in terms of comparing the pH change data after adding acid and base in

phosphate buffer and distilled water. Results showed that the change in pH was very low or

did not increase or decrease too much when using phosphate buffer. But there was a drastic

change in pH results compared to distilled water. Meaning, if the buffer is not used in a

particular solution, it tends to have a very high or very low change in pH because the presence

of the buffer resists the drastic change in pH.

REFERENCES

Butler, J. N. (1964). Ionic Equilibrium: A Mathematical Approach. Addison-Wesley. p. 151.