Laboratory Experiment

pubs.acs.org/jchemeduc

A Discovery Chemistry Experiment on Buffers

Suzanne E. Kulevich,* Richard S. Herrick, and Kenneth V. Mills
Department of Chemistry, College of the Holy Cross, Worcester, Massachusetts 01610, United States
*
S Supporting Information

ABSTRACT: The Holy Cross Chemistry Department has designed and implemented an
experiment on buffers as part of our Discovery Chemistry curriculum. The pedagogical
philosophy of Discovery Chemistry is to make the laboratory the focal point of learning for
students in their first two years of undergraduate instruction. We first pose questions in
prelaboratory sessions that serve as the focus for discussion and that guide inquiry in the lab.
We then use the experimental results to guide the lecture-hall discussion and discovery of
chemical principles. In this experiment, students first discover the Henderson−Hasselbalch
equation using pooled student data for three buffers. Students also discover the qualitative
effects of buffers by examining the influence of the addition of a strong acid, a strong base, or
water on the pH of a buffered solution versus that of nonbuffered solutions. Using their
pooled results and knowledge of titration curves discovered in the previous lab, students gain
a qualitative and quantitative understanding of buffers and report increased confidence in
making buffered solutions.
KEYWORDS: First-Year Undergraduate/General, Laboratory Instruction, Physical Chemistry, Hands-On Learning/Manipulatives,
Inquiry-Based/Discovery Learning, Acids/Bases, pH

T he Holy Cross Chemistry Department has developed a

laboratory-based guided-inquiry curriculum for our four-
semester general and organic chemistry sequence that we call
Discovery Chemistry, as described in this Journal.1−6 We have
recently reported on density, reaction kinetics, and electro-
chemistry7−9 experiments along with several organic experi-
ments.10−13 A key aspect of Discovery Chemistry is the pooling
of student data, which creates a large data set to facilitate the
use of graphical analysis. This analysis is the basis for detailed
in-class discussion and introduces students to chemistry in a
way that emphasizes the centrality of experimentation and the
inquiry-based nature of science. Successful discovery experi-
ments work best when complex topics are explored that span
multiple concepts, processes, and techniques, leading to a rich
postlaboratory discussion experience.
Buffers are a crucial concept in the curriculum for general
chemistry, analytical chemistry, and biochemistry. The
molecular explanation for buffered systems, as well as their
calculations and manipulations, are often a particularly difficult
topic for students because the understanding of buffers has
multiple levels of complexity.14 Mastering buffers requires a
strong understanding of fundamental acid−base chemistry and
chemical equilibrium, the ability to identify information
required to solve a quantitative problem, and the skill to
apply algebraic methods to compute an answer. In addition,
students often report a disconnect between practical and
theoretical understandings of the concept.14 A thorough
understanding of the derivation, utility, and limitations of the
Henderson−Hasselbalch equation, as discovered in this
experiment, helps to tie together the qualitative and
quantitative aspects of buffers.
© 2014 American Chemical Society and
Division of Chemical Education, Inc.
There are many reports of laboratory and classroom exercises
that focus on various aspects of buffers. Topics covered include
buffer capacity,15−18 buffer preparation,19,20 the Henderson−
Hasselbalch equation and it is appropriateness for use,19,21−26
analogies to promote understanding,27 classroom demonstra-
tions of buffers,28−30 and the use of biological buffers.31 One
report describes an experiential lab in which students prepare
various acetate buffer solutions and study the effect of dilution
or addition of strong acids or bases.32 Although components of
that exercise resemble our experiment described below, we
extend the analysis to multiple buffer systems covering a large
pH range, incorporate the methods of data-pooling and
graphical analysis that are central tenets of our Discovery
pedagogy, and assess student learning. In addition, we find that
employing both acidic and basic buffer systems is crucial in
demonstrating the generality of the discovery, that comparing
raw data and transformed plots is important to developing
students’ analytical skills, and that incorporating discussions of
a prior experiment on titration curves provides a comprehen-
sive learning experience for the student. We also find that
approaching the problem of buffers as a guided-inquiry exercise
increases student engagement with the material and is a more
modern pedagogical take on time-tested experiments.
We report a laboratory-based guided-inquiry experiment that
introduces students to many aspects of buffers. The experiment
builds on student understanding of chemical equilibria, acid−
base chemistry, and titration curves. Concepts that are explored
in the experiment include (1) preparing buffers, (2) buffer
range, (3) buffer capacity, (4) buffer chemistry, and (5) the law
of mass action. Processes utilized include (1) data pooling, (2)
Published: July 14, 2014
1207 dx.doi.org/10.1021/ed400377a | J. Chem. Educ. 2014, 91, 1207−1211
Journal of Chemical Education
graphing, (3) applying equations derived from theory to
experimental data, and (4) connecting data analysis with
fundamental molecular understanding of buffers. This multi-
pronged approach is applied to three buffer systems: acetic
acid/acetate, TrisHCl/Tris, and ammonium/ammonia. Stu-
dents observe and graph the properties of their assigned buffer
system, learn from pooled data how buffer systems with
different pKa values behave, and practice careful calculations
derived from acid−base equilibria. This path of learning leads
from simple observations to a comprehensive understanding of
buffers and their uses. The prelaboratory discussion and
experimental work is completed in a 4 h laboratory period.
■
PRE-LABORATORY DISCUSSION
Before beginning this Discovery Chemistry exercise, students
have studied acids and bases including strong acids and bases
and weak acid−base equilibria. In the previous week’s
experiment, pairs of students generated titration curves for
mono-, di-, or triprotic acids.33 These data serve as the focus for
subsequent lecture discussion, and representative curves are
shown as Supporting Information Figure S1. From this exercise
students discover the general shape of the titration curve and
note changes due to the strength of the acid. They learn the
meaning of pKa and how to interpolate the pKa from the
titration curve of a weak acid and, thus, estimate the relative
strength of the acid. They also observe that the pH does not
change significantly on addition of strong base until close to the
equivalence point.
In the prelaboratory discussion for the buffer experiment,
students are shown their acetic acid titration curve from the
previous week. We return to an observation about weak acid
titration curves that was not fully answered: Why does the pH
change so little in the intermediate region of the titration curve
(i.e., from about 10% to 90% of the titration of each weak acid
proton)? We then set the discovery questions, based on
observations from the preceding week: What is the nature of
the acid−base equilibrium in the buffering region, and can we
find a graphical way to more easily estimate the pKa and
strength of an acid? For the first question, students speculate
why the system is resistant to pH change in the buffering region
and if this might be different than the reason that pH changes
little after the final equivalence point is reached. We also ask
students what they might expect to observe when we add a
strong acid or base to water or to a buffered system or when we
significantly dilute a solution of acid, base, or a stoichiometric
mixture. The ideas from the students are listed on the board,
recorded by the professor, and used as the basis to begin the
postlaboratory discussions. This sets the stage for experimental
discovery, to be followed by both graphical analysis, and an
examination of the relevant acid−base equilibrium reactions
that describe buffered systems.
■
EXPERIMENTAL WORK
Students work in small groups and are assigned one of three
systems: acetic acid/acetate, TrisHCl/Tris, or ammonium/
ammonia. These are chosen so that students can learn the
generality of their observations and be able to rank the strength
of the acids. The discussion in the manual is intentionally
vague, and the provided details are only enough to allow
students to safely perform the experiment. This allows for the
student and instructor to interact and discover the concepts
together, both in the lab and in pre- and postlab discussions.
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Laboratory Experiment
First, students obtain separate 0.5 M solutions of their acid
and conjugate base. They prepare mixtures of their acid and
conjugate base, using volumetric glassware, in volume ratios of
1:1, 5:1, 10:1, 1:5, and 1:10. We believe it is good practice for
students to use volumetric glassware to make dilutions, an
important skill with which novice chemists often struggle.
However, we do not have the students make the initial
solutions, both to improve data quality and to avoid having
students handle concentrated acids and bases in an
introductory lab setting. They calibrate a Fisher Scientific
Accumet Research AR10 pH meter and record the pH of each
solution. This is a standard laboratory pH meter, and less
expensive models would likely be a good substitute.
Students are directed to use their data to plot pH as a
function of molar ratio (Supporting Information Figure S2) and
observe that there is not a linear relationship. During in-lab
conversations, they are directed to consider a plot of pH as a
function of the base-10 log of the ratio (Figure 1), which results
Figure 1. Data from pairs of students showing best-fit line using linear-
least-squares regression for ammonium/ammonia (seven pairs of
students), TrisHCl/Tris (seven pairs of students), or acetic acid/
acetate (six pairs of students). With standard error from the regression
analysis the equations of the lines are y = (0.96 ± 0.02)x + (9.17 ±
0.01), y = (0.99 ± 0.03)x + (8.16 ± 0.02), and y = (1.04 ± 0.01)x +
(4.76 ± 0.01), respectively. Students prepare mixtures of 0.5 M acid
and 0.5 M conjugate base solutions using volumetric glassware and
measure the pH of the resulting solutions.
in a linear relationship with a slope equal to one. This allows for
derivation of the Henderson−Hasselbalch equation in class,
both by graphical analysis and by derivation of the equation
from the Ka expression. In-class lectures use this equation to
help students learn how to prepare buffers and how to evaluate
the relative population of acid and conjugate base using these
three acids as examples. (Instructors also discuss the limitations
of the Henderson−Hasselbalch equation in lecture, drawing on
examples from the titration curve analysis from the previous lab
where the small x approximation is not valid.)
In the second part of the experiment, students add
concentrated HCl or NaOH to water and to a 1:1 molar
mixture of their acid/conjugate base pair (Figure 2). They
record the initial and final pH of each solution. They plot the
data as instructed, creating separate lines for the water and
buffer solutions. If instructors have time, they might consider
testing different ratios of acid/conjugate base pairs to help
discover the concept of buffer capacity.
dx.doi.org/10.1021/ed400377a | J. Chem. Educ. 2014, 91, 1207−1211
Journal of Chemical Education Laboratory Experiment

We conducted an assessment quiz before the initial prelab

discussion and before the prelab discussion the following
week.34 The questions and student responses are reported in
Table 1.

Table 1. Student Assessment Data from a Before and After

Survey, 2013
Question Answersa Prelabb,c Postlabc
1. On 10-fold dilution of a strong Increase 43% 84%
acid, the pH will:
Decrease 54% 11%
Stay about the same 3% 5%
2. On 10-fold dilution of a buffered Increase 10% 4%
solution, the pH will:
Decrease 11% 10%
Figure 2. Data from seven pairs of students showing the addition of Stay about the 79% 87%
concentrated HCl or NaOH to a stoichiometric mixture of same
ammonium/ammonia or to distilled water. Lines are added to guide 3. If one adds a small amount of Increase slightly 73% 82%
the eye. The initial pH of water is reported as 7 for all student pairs. strong base to a buffered solution,
Representative data for the other two systems are given as Supporting the pH will:
Information Figure S3. Students make two separate 20 mL solutions Increase by a large 5% 4%
by combining 10 mL of 0.5 M acid and 10 mL of 0.5 M conjugate amount
base. They measure the pH of each solution, then add 10 mL of 0.1 M Decrease slightly 16% 16%
HCl to one solution and 10 mL of 0.1 M NaOH to the other, mix, and Decrease by a large 6% 0%
measure the pH again. They also add 10 mL of 0.1 M HCl to 20 mL of amount
water in one beaker and 10 mL of 0.1 M NaOH to 20 mL of water in 4. To have a solution that is A weak acid/con- 43% 71%
considered a buffer, one must have jugate base pair
another beaker and measure the pH of each.
A strong acid/con- 30% 19%
jugate base pair
A mixture of a 27% 11%
strong acid and
In the third part of lab, students perform 10- and 100-fold strong base
serial dilutions of their solutions of acid, conjugate base, or the 5. The pH of a buffer solution Is always equal to 9% 1%
stoichiometric mixture. We find that students often struggle pH 7
with dilutions both conceptually and in the lab and can benefit Depends on the pKa 7% 8%
of the acid
from a review of this useful laboratory technique. Students Depends on the 11% 14%
measure the pH of each initial solution and after each of the ratio of base to
dilutions and graph their data as in Figure 3. acid
Both b. and c. 72% 77%
6. How confident are you that you Not very confident 61% 4%
could create a buffer?
Somewhat confident 31% 40%
Confident 7% 45%
Very confident 1% 11%
a
Correct answer in bold. bMost popular answer in bold italics for pre-
and postlab surveys. cNumbers might not equal 100% due to rounding.
140−143 respondents per question.

Figure 3. Data from seven pairs of students showing the influence of
dilution on a solution of ammonia, ammonium, or a stoichiometric
mixture. Students are instructed to connect their single measurements
to guide the eye; the lines above connect a point of the average value
for each measurement. Representative data for the other two systems
are given as Supporting Information Figure S4. Students add 10 mL of
0.5 M acid to 10 mL of 0.5 M conjugate base and mix. They pipet 5
mL of this solution into a 50 mL volumetric flask and bring to volume
with water. They transfer this solution to a beaker and repeat the 10-
fold dilution. The same two dilutions are performed with the 0.5 M
acid and 0.5 M conjugate base solutions, and the pH of each solution is
measured with the pH meter.
1209
■ HAZARDS
Acetic acid (CAS # 64-19-7) is corrosive and flammable;
sodium acetate (CAS # 127-09-3), ammonium chloride (CAS #
12135-02-9), Tris (CAS # 77-86-1), and Tris−HCl (CAS #
1185-53-1) are irritants; aqueous ammonia (CAS # 7664-41-7)
is toxic, corrosive, and a lachrymator; aqueous HCl (CAS #
76447-01-0) is corrosive and toxic. Sodium hydroxide (CAS #
1310-73-2) is corrosive, causes skin burns, and is harmful or
fatal if swallowed or inhaled. Safety glasses and gloves must be
worn.
■ POSTLAB DISCUSSION (INTERPRETATION OF
RESULTS)
This experiment is well received by students, mostly likely due
to its experiential learning element.35 Students discover the
Henderson−Hasselbalch equation by making and testing buffer
solutions of various component ratios rather than simply
dx.doi.org/10.1021/ed400377a | J. Chem. Educ. 2014, 91, 1207−1211
Journal of Chemical Education
memorizing an equation. By plotting their results, they observe
mathematical trends and see linear relationships. They discover
that buffer solutions resist pH change when diluted or treated
with strong acid or base while working with modern digital pH
meters.
Via this concrete, hands-on experience, students observe that
although buffered systems change pH with manipulation, they
resist large changes in pH. Subsequent lectures begin with
reflective observation and remind students of the qualitative
aspects of buffers. These ideas are reinforced by examination of
the pooled data and graphs for the three buffer systems.
As part of learning how to manipulate equations, the
discussion returns to the weak acid titration curves from the
previous week’s experiment. During that titration experiment,
students were told that the pKa can be estimated by the pH at
the half-equivalence point for conditions under which the small
x approximation is valid and interpolated from their curve to
estimate the pKa and the relative strength of the acid. This
week, using the plot that allows them to discover the
Henderson−Hasselbalch equation, students learn how to use
an equation based on a linear best-fit line to more accurately
calculate the pKa.
This week’s data also allow students to interpret the shape of
their weak acid titration curves in the following four areas. First,
using the previous week’s data, students observe that the pH
does not vary much in the region near the half-equivalence
point. During the discussions that follow the buffers lab, we
return to these titration curves and discuss what species were in
solution in these regions and why they would or would not
make a buffer. We also make reference to our titration curve
with HCl and ask students to consider why, although the pH
does not change near the half-equivalence point, this solution
does not constitute a buffer. As part of this thought exercise,
students are asked to write the balanced equations for both the
neutralization and resultant equilibrium reactions for strong
and weak acids.
Second, the combination of the two weeks of laboratories
also allows for a graphical appreciation of buffer capacity. We
connect our lecture discussion about buffer capacity to the
observation of the curvature of the titration curve outside of the
buffering range. Students learn why the base to conjugate acid
ratios of 1:10 and 10:1 are commonly used as the lower and
upper limits for use of the Henderson−Hasselbalch equation.
Third, from the dilution experiment, we discuss the balance
between the concentration of the buffer and its ability to resist
changes in pH. This allows for a segue into methods for
preparation of a buffer, noting that students have prepared
buffers in both weeks: titrating a weak acid with a strong base in
week one when making a titration curve and mixing solutions
of acid and conjugate base in week two. Finally, we link the
regions of the titration curves that students find most
surprising, such as the shallow inflection points for relative
strong weak acids or conjugate bases, with the limitations of the
assumptions behind the Henderson−Hasselbalch equation.
This period of discernment is important in building a
foundation that is then used for abstract conceptualization. In
this last crucial phase of discussion, students learn how to
manipulate chemical equilibria to generate equations that
explain the quantitative results obtained in lab. Students are
expected to be able to write all of the relevant chemical
equations that describe both the neutralization and subsequent
equilibrium reactions and connect these symbolic chemical
equations to the quantitative relationships they observed
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Laboratory Experiment
empirically. They see that the linear relationship obtained is
based on the Henderson−Hasselbalch equation, and they
discover how to use the equation to answer questions about
buffers. We also direct discussions using balanced chemical
equations that require students to reflect on the relationship
between Le Chatelier’s principle, the common ion effect, and
buffered systems. These concepts are subsequently reinforced
the following week during a lab that utilizes solubility and
complex-ion formation equilibria. The concepts and their
applications are also reinforced with follow-up problem sets and
exam questions that focus on the analysis of lab data, the use of
the chemical concepts to perform practical calculations relating
to pH prediction and buffer preparation, and more in-depth
thought questions that require students to apply what they have
learned in the lab and lecture to new and more complex
problems on their own.
■ CONCLUSION
Our assessment data suggest that when students prepare and
manipulate buffers and make thoughtful use of graphical
analysis, their understanding of topics related to acid−base
equilibrium and their confidence with preparing buffered
systems improve (Table 1). Using three separate systems
allows students to make discoveries with their own data and
generalize with class data. The lab experience creates both a
concrete and a qualitative picture of the concept of a buffer.
This reflective discovery allows students to tackle difficult buffer
questions (see Supporting Information) and apply their
qualitative and quantitative understanding of buffered systems
to future courses.
■
ASSOCIATED CONTENT
*
S Supporting Information
Student lab manual; instructor prep notes, including CAS
numbers and hazards; student data sheet; sample postlab
follow-up exercises and exam questions; Figures S1−S4. This
material is available via the Internet at http://pubs.acs.org.This
material is available via the Internet at http://pubs.acs.org.
■
AUTHOR INFORMATION
Corresponding Author
*E-mail: skulevic@holycross.edu.
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
We gratefully acknowledge the assistance of Jane Jennette,
Jamie Herrick, Maria Fistik, and Elizabeth Landis. This material
is based upon work supported by the National Science
Foundation under grants MCB-0950245 and MCB-1244089
(K.V.M.), by a Henry Dreyfus Teacher-Scholar Award
(K.V.M.), and by a Petroleum Research Fund grant 51085-
UR3 (R.S.H.).
■
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