Name: REY P. AMISTOSO CHEM.

27A-DVM2

Instructor: MS. ELLEN JOY P. PANDAN Date Submitted: March 11, 2019

FACTORS AFFECTING BUFFER CAPACITY

(pH and Buffer System)

I. Introduction

A buffer is a solution that can resist pH change upon the addition of an acidic or basic components. It
is able to neutralized small amounts of added acid or base, maintaining the pH relatively stable. Buffer
solutions can be made to maintain almost any pH, depending on the acid-base pair used. The pH of a
buffer solution is determined by the Ka of the acid and by the ratio of concentrations of [HA] and [A–]. The
expression for the Ka of the conjugate acid of the buffer:
[𝑯+][𝑨−] [𝑯𝑨]
Ką= [𝑯𝑨]
BECOMES [H+]=Ką
[𝑨−]

Buffers are prepared by combining a weak acid and its conjugate (acetic acid and sodium acetate).
Buffers are prepared by combining a weak acid and its conjugate (acetic acid and sodium acetate). In
general, the pH range in which a buffer solution is effective at +/- one pH unit on either side of the pKa.
The Henderson–Hasselbalch provides the information needed to prepare a buffer.

𝒄𝒐𝒏𝒋𝒖𝒈𝒂𝒕𝒆 𝒃𝒂𝒔𝒆 [𝑨−]

pH= pKą+log [ ]
𝒘𝒆𝒂𝒌 𝒂𝒄𝒊𝒅 [𝑯𝑨]

In this experiment, the Henderson-Hasselbalch equation will be used to determine the amount of
acetic acid and sodium acetate required to prepare a series of buffer solutions. Once the buffer solutions
have been prepared, their buffer capacity will be determined. A buffer solution can maintain its
approximate pH when an acid or a base is added to it because it can react with both acids and bases. If a
strong acid (H3O+) is added, the basic component of the buffer (A–) can react with it, and if a strong base
(OH–) is added, the acidic component of the buffer (HA) will react with it.

The buffer will be prepared by choosing the appropriate acid-base pair, calculating the molar ratio
of acid to base that will produce the assigned pH, and then mixing the calculated amounts of the two
compounds with enough deionized/distilled water to make a buffer solution.

A solution with approximately the same pH as the buffer solution will be prepared by diluting a
solution of a strong acid or a strong base. This solution will not be a buffer solution, as can be shown by
comparing its buffering ability to that of the buffer solution. Finally, a known amount of strong acid or
base will be added to the buffer solution. Before adding the acid or base, the pH change that the addition
should cause will be calculated. The observed pH change will be compared to the calculated value. There
are many different factors that play a role in choosing a particular buffer system for a particular biological
application. These factors include working temperature, desired pH, toxicity to the system, and
interactions with other components. Interactions with other components was the factor investigated in
this experiment.

To testify and verify the above concept and idea, an experiment was conducted with the
following objectives:

1. To prepare various buffer solutions at a specific pH and to determine their buffer

capacity.
2. To learn to identify solutions that are buffers
3. To understand the how a buffer resists changes in pH upon addition of acid or base
solutions and;
4. To determine the effect of the following on buffer capacity:
a. Concentration of the buffer
b. Ratio of the conjugate base to the weak acid

II. Methodology

A. Chemicals, Apparatus and Materials

The chemicals used in the experiment are as follows: 0.1 M acetic acid (CH3COOH), 1 M sodium
acetate (NaCH3COO), 0.1 M sodium acetate (NaCH3COO), 0.1 N sodium hydroxide (NaOH), 0.1 N
hydrochloric acid (HCl).

The apparatus used in the experiment include pH meter, beaker, graduated cylinder, Erlenmeyer
flask, volumetric flask, pipette, aspirator, wash bottle with distilled water, stirring rod and two 500-mL
empty bottles.

B. Procedure

Effect of buffer concentration on buffer capacity

In preparation of acetic-acid buffer solution with the pH of 4.7 for buffers 1 and 2. We calculated
first the volume ratio of 1.0 M acetic acid and 1.0 M sodium acetate using the Henderson-Hasselbalch
equation. For buffer 1, we prepared 250-mL of 0.10 M of acetic acid-acetate buffer and for buffer 2 we
prepared 100-mL of 0.05 M acetic acid-acetate buffer which has the 4.76 pK𝑎 of acetic acid. In buffer 3,
in order to prepare the buffer 3 we calculated the volume needed to prepare 100 mL of 0.005 M acetic
acid-acetate buffer solutions with pH of 4.7 from 0.10 M of acetic acid-acetate by using the dilution of
𝐶2𝑉2
solution with formula of V1= .
𝐶1
 After we prepared buffers 1,2 and 3, we placed 55-mL of buffer 1 solution into the 100-mL beaker.
Then we measured the pH with the use of pH meter electrode and record the result pH in the table 1 of
the data sheet. Subsequently, we added 2 mL of 0.1 NaOH next, we stirred the mixture and measure the
pH. At that time, repeat steps 5, 6, and 7 for buffers 1 and 2. Then, we calculated the difference in the pH
values before and after addition of NaOH and we compared the magnitude of the change in pH among
Buffers 1, 2 and 3.

Effect of the ratio of the conjugate base to the weak acid on buffer capacity

In initial procedure, we calculated the volume needed to prepare the following solutions by
dilution. Afterwards, from the Henderson-Hasselbalch equation, we calculated the volume ratio of acetic
acid and sodium acetate ratio to produce buffer solution with pH 3.76, pH 4.76 and pH 5.76. Using 0.1 M
acetic acid (CH3COOH) and 0.1 M sodium acetate (NaCH3COO), make op 110 mL of the buffer solution.
After that, we measured and record the pH of the buffer solution in the table 2 (before addition of NaOH)
and in table 3 (before addition of HCl) on the data sheet. We divided the buffer into to two equal and
separate parts of 100-mL beakers. We added 2 mL of 0.1 N NaOH to the first beaker and 2 mL of 0.1 N HCl
on the second beaker. Eventually, we recorded the pH of each buffer solution after addition of alkali or
acid. The first result of the first beaker which is the addition of NaOH was recorded on the table 2 on the
data sheet and other one was on the table of 3 of the data sheet. Make the buffers 5 and 6 in a manner
similar in step 5, 6 and 7 and we calculated the difference in the pH values before and after the addition
of NaOH (table 2) and HCl (table 3). We also compared the magnitude of the change in pH among buffers
4, 5, and 6.

III. Results and Discussion

Buffer 1: 250-mL of 0.10 M acetic acid-acetate buffer

𝟎.𝟖𝟕 𝟏
[CH3COO-] = 𝟏.𝟖𝟕 = 𝟎. 𝟒𝟔𝟓 [CH3COOH] = 𝟏.𝟖𝟕 = 𝟎. 𝟓𝟑𝟒

Volume of [CH3COO-] in mL Volume of [CH3COOH] in mL

𝟎.𝟒𝟔𝟓 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟐𝟓 𝑳 𝟎.𝟓𝟑𝟒 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟐𝟓 𝑳
[CH3COO-]= 𝟏𝑳 = 𝟏𝟏. 𝟔 𝒎𝑳 [CH3COOH] = 𝟏𝑳 = 𝟏𝟑. 𝟒 𝒎𝑳
𝟏.𝟎 𝑴 𝒙 𝟏.𝟎 𝑴 𝒙
𝟏𝟎𝟎𝟎 𝒎𝑳 𝟏𝟎𝟎𝟎 𝒎𝑳

Buffer 2: 100 mL of 0.05 M acetic acid-acetate buffer

𝟎.𝟖𝟕 𝟏
[CH3COO-] = 𝟏.𝟖𝟕 = 𝟎. 𝟒𝟔𝟓 [CH3COOH]= 𝟏.𝟖𝟕 = 𝟎. 𝟓𝟑𝟒

Volume of [CH3COO-] in mL Volume of [CH3COOH] in mL

𝟎.𝟒𝟔𝟓 𝒙 𝟎.𝟎𝟓 𝑴 𝒙 𝟎.𝟏 𝑳 𝟎.𝟓𝟑𝟒 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟏 𝑳
[CH3COO-]= 𝟏𝑳 = 𝟐. 𝟑 𝒎𝑳 [CH3COOH]= 𝟏𝑳 = 𝟐. 𝟕 𝒎𝑳
𝟏.𝟎 𝑴 𝒙 𝟏.𝟎 𝑴 𝒙
𝟏𝟎𝟎𝟎 𝒎𝑳 𝟏𝟎𝟎𝟎 𝒎𝑳
Buffer 3: 100 mL of 0.005 M acetic acid-acetate buffer solution from 0.10 M acetic acid-acetate

C1= 0.10 M V1=? C2= 0.005 M V2= 100 mL

𝑪𝟐𝑽𝟐 (𝟎.𝟎𝟎𝟓 𝑴)(𝟏𝟎𝟎 𝒎𝒍)
V1= = = 𝟓 𝒎𝑳
𝑪𝟏 𝟎.𝟏𝟎 𝑴

Table 1: Effect of Buffer Concentration on Buffer Capacity

Concentration Actual reading pH

Buffer System (M) Before Addition of After Addition of Change in pH
NaOH NaOH
Buffer 1 0.10 4.76 4.79 0.03
Buffer 2 0.05 4.75 4.94 0.19
Buffer 3 0.005 4.32 4.57 0.25

 250-mL of 0.1 M of acetic acid solution from 1.0 M stock solution of acetic acid
C1= 1.0 M V1=? C2= 0.1 M V2= 250 mL

𝑪𝟐𝑽𝟐 (𝟎.𝟏 𝑴)(𝟐𝟓𝟎 𝒎𝑳)

V1= = = 𝟐𝟓 𝒎𝑳
𝑪𝟏 𝟏.𝟎 𝑴

 250-mL of 0.1 M sodium acetate solution from 1.0 stock solution of sodium acetate
C1=1.0 M V1=? C2= 0.1 M V2= 250 mL
𝑪𝟐𝑽𝟐 (𝟎.𝟏 𝑴)(𝟐𝟓𝟎 𝒎𝑳)
V1= = = 𝟐𝟓 𝒎𝑳
𝑪𝟏 𝟏.𝟎 𝑴

 Using the Henderson-Hasselbalch equation, calculate the volume ratio of 0.1 M acetic acid and
0.1 M sodium acetate solutions needed to prepare 110 mL each of the following buffer solutions.

Buffer 4: pH= 3.76

[𝑪𝑯𝟑𝑪𝑶𝑶−] 𝟎.𝟏 𝟏
= 𝟏𝟎𝟑.𝟕𝟔−𝟒.𝟕𝟔 = 𝟎. 𝟏 [CH3COO-]= = 𝟎. 𝟎𝟗𝟏 [CH3COOH]=𝟏.𝟏 = 𝟎. 𝟗𝟎𝟗
[𝑪𝑯𝟑𝑪𝑶𝑶𝑯} 𝟏.𝟏

Volume in [Ch3COO-] in mL Volume in [CH3COOH] in mL

𝟎.𝟎𝟗𝟏 𝒙 𝟎.𝟏 𝑴 𝒙 𝟎.𝟏𝟏 𝑳 𝟎.𝟗𝟎𝟗 𝒙 𝟎.𝟏 𝑴 𝒙 𝟎.𝟏𝟏 𝑳
[CH3COO-]= 𝟏𝑳 = 𝟏. 𝟎𝟎𝟏 𝒎𝑳 [CH3COOH]= 𝟏𝑳 = 𝟏𝟎 𝒎𝑳
(𝟏.𝟎 𝑴) (𝟏.𝟎 𝑴)
𝟏𝟎𝟎𝟎 𝒎𝑳 𝟏𝟎𝟎𝟎 𝒎𝑳

Buffer 5: pH= 4.76

[𝑪𝑯𝟑𝑪𝑶𝑶−] 𝟏 𝟏
= 𝟏𝟎𝟒.𝟕𝟔−𝟒.𝟕𝟔 = 𝟏 [CH3COO-] = 𝟐 = 𝟎. 𝟓 [CH3COOH] = 𝟐 = 𝟎. 𝟓
[𝑪𝑯𝟑𝑪𝑶𝑶𝑯]

Volume of [CH3COO-] in mL Volume of [CH3COOH] in mL

 𝟎.𝟓 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟏𝟏 𝑳 𝟎.𝟎𝟗𝟏 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟏𝟏 𝑳
[𝑪𝑯𝟑𝑪𝑶𝑶 −] = 𝟏𝑳 = 𝟓. 𝟓 𝒎𝑳 [CH3COOH]= 𝟏𝑳 = 𝟓. 𝟓 𝒎𝑳
(𝟏.𝟎 𝑴) (𝟏.𝟎 𝑴)
𝟏𝟎𝟎𝟎 𝒎𝑳 𝟏𝟎𝟎𝟎 𝒎𝑳

Buffer 6: pH= 5.76

[𝑪𝑯𝟑𝑪𝑶𝑶−] 𝟏𝟎 𝟏
= 𝟏𝟎𝟓.𝟕𝟔−𝟒.𝟕𝟔 = 𝟏𝟎 [CH3COO-] =𝟏𝟏 = 𝟎. 𝟗𝟎𝟗 [CH3COOH] = 𝟏𝟏 = 𝟎. 𝟎𝟗𝟏
[𝑪𝑯𝟑𝑪𝑶𝑶𝑯]

Volume of [CH3COO-] in mL Volume of [CH3COOH] in mL

𝟎.𝟗𝟎𝟗 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟏𝟏 𝑳 𝟎.𝟎𝟗𝟏 𝒙 𝟎.𝟏𝟎 𝑴 𝒙 𝟎.𝟏𝟏 𝑳
[CH3COO-] = 𝟏𝑳 = 𝟏𝟎 𝒎𝑳 [CH3COOH]= 𝟏𝑳 = 𝟏. 𝟎𝟎𝟏 𝒎𝑳
(𝟏.𝟎 𝑴) (𝟏.𝟎 𝑴)
𝟏𝟎𝟎𝟎 𝒎𝑳 𝟏𝟎𝟎𝟎 𝒎𝑳

Table 2: Effect of the ratio of the conjugate base to the weak acid, [A-]/[HA], on buffer capacity upon
addition of NaOH

Concentration Actual pH Reading

Buffer System (M) Before addition After addition Change in pH
NaOH of NaOH
Buffer 4 3.76 3.46 3.81 0.35
Buffer 5 4.76 3.63 4.76 0.13
Buffer 6 5.76 5.67 5.80 0.13

The reaction responsible for maintaining constant pH on the addition of acid or base in buffer
solution are called buffer action. The buffer action of acetic acid and sodium acetate mixture can be
explained as follows.

CH3COOH + H2O CH3COO- + H3O+

When a strong base such as NaOH is added to the buffer, the weak acid will give up its H+ in order
to transform the base (OH-) into water (H2O) and the conjugate base: HA + OH- → A- + H2O. The acetic acid
reacts with and consumes the excess OH- ion. The hydrogen ion concentration decreases by less than the
amount expected for the quantity of base added. The OH- reacts with the H3O+ ion from the acid in the
following reaction:

H2O + CH3COOH H3O+ + CH3COO-

H3O+ + OH- H2O
The OH- reduces the H3O+ ion concentration, which causes a shift to the right, forming additional
CH3COO- and H3O+ ions. For each mole of OH- added consumes a mole of CH3COOH and produces mole
of CH3COO-.

OH- + CH3COOH CH3COO- + H2O

Table 3: Effect of the ration of the conjugate base to the weak acid, [A-]/[HA], on buffer capacity upon
addition of HCl

Actual pH Reading
Buffer System Calculated pH Before addition After addition Changes in pH
of HCl of HCl
Buffer 4 3.76 3.42 2.79 0.66
Buffer 5 4.76 4.63 4.49 0.14
Buffer 6 5.76 5.68 5.57 0.11

When a strong acid such as HCl is added to the buffer, the hydronium ions react with the CH3COO-
ions of the salt and form more undissociated CH3COOH and produces few H+ ions in solution and therefore
only a little change in pH.

H3O+ + CH3COO- CH3COOH + H2O

There is a limit to the quantity of H+ or OH- that a buffer can absorb without undergoing a
significant change in pH. If a mole of HCl to a liter of buffer solution of sodium acetate/ acetic acid buffer
the H+ completely consumes the buffer and results in a drastic change in pH.

When the pH was closer to the pKa, the concentrations of the weak acid and its conjugate base
were closer to the equimolar point respect to other solutions which pH values were not so close to the
pKa. In detail, when HCl was added to the solutions, the conjugate base started to receive its proton, so
that the pH fluctuation was controlled until the conjugate base molecules finished as proton acceptors. In
this point, the protons were able to dissociate in the solution and modify the pH (acidized). When the pH
is closer to the pKa the concentration of protons decreases; when the pH is lower than the pKa, the
concentration of protons increases and saturates the conjugate base.

Regarding to the experiment, the buffer capacity refers to the amount of added acid or added
base that can be neutralized by a buffer. It is determined by the concentrations of the conjugate base and
conjugate acid. The higher the concentration the greater the buffer capacity. The more A- and HA
molecule available, the less of an effect addition of a strong acid or base will have on the pH system. When
the ratio is closer to 1 the greater the buffer capacity. A buffer is most effective when the concentrations
of acid and conjugate base are approximately equal. Therefore, the more ratio needs to differ to desire
pH, the less effective the buffers.
 IV. Conclusion

As for the conclusions, A buffer is a mixture of a weak acid and its conjugate base because the
solute can either absorb protons or release protons, the buffer solution can maintain the pH at a constant
value. They act to moderate gross changes in pH. We prepared a buffer of an assigned pH value. We also
determine the buffering ability as measured by the buffer capacity. The ability to resist pH change depends
on the concentrations of the acid and conjugate base present. In general, the more concentrated the
buffer, the higher the buffer capacity, as more conjugate acid and conjugate base are present to react
with any H+ or OH- added to the buffer. The ability to resist changes in pH also depends on the pH of the
buffer as compared to the pKa of the weak acid used to prepare it. At pH values below the pKa, the buffer
will have a higher buffer capacity for additional OH- than for H+, as the concentration of the conjugate acid
will be higher than that of the conjugate base. The reverse is true for buffers with pH values above the
pKa. The further the pH is from the pKa, the more extreme are the differences.

To avoid error in this activity or experiment, before using any apparatus. it is necessary to clean it
up first. This is to ensure that the apparatus is free contamination. The cleaning processes have to take
charge while handling the pH electrode as well. This is a must in order to obtain a desirable pH value. A
possible source of error was the sodium acetate and acetic acid solutions preparation. The acetic acid was
at 97% in the stock, whereas sodium acetate was pure and solid. When preparing the sodium acetate
solution, probably, some of it stayed in the container and was not added to the buffer solution, decreasing
the concentration of conjugate base, increasing the concentration of acid and hence, decreasing pH values
in the solutions.
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Brown T. L, LeMay, H. E. Jr., Burstein, B. E. (2012). Chemistry. The Central Science (12th edition).
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Harris, D.C. Quantitative Chemical Analysis, 7th ed.; W.H. Freeman & Co.: New York, 2007; pp 53-59 and
167-176.

Devlin, T. M. Textbook of Biochemistry With Clinical Correlations (3rd ed.). New York: Wiley-Liss, 1992. Pp.
6-13, 34-41.
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