BIOCHEMISTRY
LABORATORY
Activity 2.
pH and Buffer
Solutions
OBJECTIVES:
At the end of the exercise, the students should be able to:
1. Demonstrate how to measure pH of a solution using indicators.
2. Determine the operations of buffer systems.
3. Compare the accuracy of pH meter and pH paper.
4. Calculate the pH given the concentration of H+ or concentration of OH-
5. Predict the acidity or alkalinity from the given concentration.
INTRODUCTION
Buffers are systems that resist a change in pH even when we add acids or bases
to it. This system is a mixture of a weak acid and its conjugate base or a mixture
of a weak base and its conjugate acid. In our blood, carbonic acid is the most
important buffer. This solution maintains our blood pH (7.35-7.45) to facilitate
transport of oxygen from the lungs to the cells. The ability of buffers to resist
changes in pH is ruled by le Chatellier’s principle- principle of equilibrium shift
due to changes in buffer conditions.
MATERIALS/EQUIPMENT
• 12 test tubes with rack
• pH paper
• wash bottle
• eight 50 mL beakers
• two 10 mL graduated cylinder
• two 10 mL pipettes
REAGENTS AND SAMPLES
0.1 M hydrochloric acid
0.1 M acetic acid
0.1 M sodium acetate
0.1 M carbonic acid
0.1 M sodium bicarbonate
0.1 M ammonia
0.1 M sodium hydroxide
PROCEDURE
A. Measurement of pH
1. Get 7 test tubes.
2. Put 1mL of 0.1M HCl in the first test tube and dip the pH paper.
3. Repeat the procedure with 0.1M acetic acid, 0.1M sodium
acetate, 0.1M carbonic acid, 0.1M sodium bicarbonate, 0.1M ammonia,
and 0.1M sodium hydroxide.
4. Record your results on report sheet.
If the pH meter is available, determine the pH of the solutions listed above.
Record your results and compare them with the result you obtained in using the
pH paper.
B. Buffer System
Get four 50 mL beakers. Label them as A,B,C and D. Prepare buffer systems in
four beakers as follows.
1. 10 mL 0.1M CH3COOH + 10 mL M 0.1NaC2H3O2
2. 5 mL 0.1M CH3COOH + 15 mL M 0.1NaC2H3O2
3. 10 mL 0.1 M H2CO3+ 10 mL 0.1 NaHCO3
4. 5 mL 0.1 M H2CO3+ 15 mL 0.1 NaHCO3
Measure the pH of each buffer system using either pH paper or pH meter.
Record your results. Divide each of the buffers (A-D) into halves (10mL each) and
place half of each in a clean, dry 50 mL beaker. To the first half, add 1 mL of 0.1
M HCl. Mix and determine the pH. To the second half add 1 mL of 0.1 M NaOH.
Mix and determine the pH. Record your answers.
Guide Questions:
1. Which gives more pH value, pH meter or pH paper? Explain.
-pH meter gives more pH value than pH paper because it has a digital
interface, by using standarlized buffer which allow the meter to associate a
particular voltage with a pH value. The result can be given on a digital displays
which will provide more precise result than the pH paper.
Guide Questions:
2. Which of the buffers prepared had the best buffer capacity?
-The soap had the best buffer capacity, which reacted into red and blue
litmus paper. In red litmus paper it turns out into blue while in blue litmus paper
it has no reaction. And base from that situation we concluded that the soap
solution is a base where in, we found out that it has pH 9-10 compared to the
other solution that only have pH2.
Guide Questions:
3. A buffer solution contains acetic acid and sodium acetate. The concentration
of each substance is 0.50 M. determine the pH of the solution (Ka= 1.8 x 10-5).
Guide Questions:
4. Cite some applications of a buffer solution in the following:
a. Food
b. Photography
c. Agriculture
a. Food - In a buffer solution, through the acid and base (acidity and basicity), it
measures the substances of free acidity that help to digest foods and that
maintain health, usually we eat are acidic because pH is also in maintaining.
b. Photography - Buffer solution also helps photography to determine and
capture good memories of pictures. It helps to develop some processes to make
good photography.
c. Agriculture - Through the pH of neutral, it helps the soil to grow effectively
and maintaining having a good formation and production of glomalin. It helps to
control proper food processing and regulates by having nutrients in the soil.
LEADER: MAYWAY BULAO
MEMBERS:
ANGELICA MARTINEZ
VICAH JOY CORPUZ
IVY ANN NICOLE LEAÑO
ABEGAIL MAMBA
RHENALYN BAUTISTA
