Chapter 3

The Vocabulary
of Analytical Chemistry
Chapter Overview
3A Analysis, Determination, and Measurement
3B Techniques, Methods, Procedures, and Protocols
3C Classifying Analytical Techniques
3D Selecting an Analytical Method
3E Developing the Procedure
3F Protocols
3G The Importance of Analytical Methodology
3H Key Terms
3I Chapter Summary
3J Problems
3K Solutions to Practice Exercises

If you browse through an issue of the journal Analytical Chemistry, you will discover that the
authors and readers share a common vocabulary of analytical terms. You probably are familiar
with some of these terms, such as accuracy and precision, but other terms, such as analyte and
matrix, are perhaps less familiar to you. In order to participate in any community, one must first
understand its vocabulary; the goal of this chapter, therefore, is to introduce some important
analytical terms. Becoming comfortable with these terms will make the chapters that follow
easier to read and to understand.

41
42 Analytical Chemistry 2.1

3A Analysis, Determination and Measurement

The first important distinction we will make is among the terms analysis,
determination, and measurement. An analysis provides chemical or physi-
cal information about a sample. The component in the sample of interest
to us is called the analyte, and the remainder of the sample is the matrix.
In an analysis we determine the identity, the concentration, or the proper-
ties of an analyte. To make this determination we measure one or more of
the analyte’s chemical or physical properties.
An example will help clarify the difference between an analysis, a de-
termination and a measurement. In 1974 the federal government en-
acted the Safe Drinking Water Act to ensure the safety of the nation’s public
drinking water supplies. To comply with this act, municipalities monitor
their drinking water supply for potentially harmful substances, such as
fecal coliform bacteria. Municipal water departments collect and analyze
samples from their water supply. To determine the concentration of fecal
A fecal coliform count provides a gen-
eral measure of the presence of patho- coliform bacteria an analyst passes a portion of water through a membrane
genic organisms in a water supply. For filter, places the filter in a dish that contains a nutrient broth, and incubates
drinking water, the current maximum the sample for 22–24 hrs at 44.5 oC ± 0.2 oC. At the end of the incuba-
contaminant level (MCL) for total co-
liforms, including fecal coliforms is less tion period the analyst counts the number of bacterial colonies in the dish
than 1 colony/100 mL. Municipal water and reports the result as the number of colonies per 100 mL (Figure 3.1).
departments must regularly test the water Thus, a municipal water department analyzes samples of water to determine
supply and must take action if more than
5% of the samples in any month test posi- the concentration of fecal coliform bacteria by measuring the number of
tive for coliform bacteria. bacterial colonies that form during a carefully defined incubation period.

Figure 3.1 Colonies of fecal coliform bacteria from a water supply. Source: Susan
Boyer. Photo courtesy of ARS–USDA (www.ars.usda.gov).
 Chapter 3 The Vocabulary of Analytical Chemistry 43

3B Techniques, Methods, Procedures, and Protocols

Suppose you are asked to develop an analytical method to determine the
concentration of lead in drinking water. How would you approach this
problem? To provide a structure for answering this question, it is helpful to
consider four levels of analytical methodology: techniques, methods, pro-
cedures, and protocols.1
A technique is any chemical or physical principle that we can use
to study an analyte. There are many techniques for that we can use to de-
termine the concentration of lead in drinking water.2 In graphite furnace See Chapter 10 for a discussion of graph-
ite furnace atomic absorption spectrosco-
atomic absorption spectroscopy (GFAAS), for example, we first convert py. Chapters 8–13 provide coverage for a
aqueous lead ions into free atoms—a process we call atomization. We then range of important analytical techniques.
measure the amount of light absorbed by the free atoms. Thus, GFAAS uses
both a chemical principle (atomization) and a physical principle (absorp-
tion of light).
A method is the application of a technique for a specific analyte in a
specific matrix. As shown in Figure 3.2, the GFAAS method for determin-
ing the concentration of lead in water is different from that for lead in soil
or blood.
A procedure is a set of written directions that tell us how to apply a
method to a particular sample, including information on how to collect the
sample, how to handle interferents, and how to validate results. A method
may have several procedures as each analyst or agency adapts it to a specific
need. As shown in Figure 3.2, the American Public Health Agency and
1 Taylor, J. K. Anal. Chem. 1983, 55, 600A–608A.
2 Fitch, A.; Wang, Y.; Mellican, S.; Macha, S. Anal. Chem. 1996, 68, 727A–731A.

Graphite Furnace Atomic Absorption Spectroscopy

Techniques
(GFAAS)

Methods Pb in Soil Pb in Water Pb in Blood

Figure 3.2 Chart showing the hierar-

chical relationship between a technique,
Procedures APHA ASTM methods that use the technique, and
procedures and protocols for a method.
The abbreviations are APHA: Ameri-
can Public Health Association, ASTM:
Protocols American Society for Testing Materi-
EPA als, EPA: Environmental Protection
Agency.
44 Analytical Chemistry 2.1

the American Society for Testing Materials publish separate procedures for
determining the concentration of lead in water.
Finally, a protocol is a set of stringent guidelines that specify a pro-
cedure that an analyst must follow if an agency is to accept the results.
Protocols are common when the result of an analysis supports or defines
public policy. When determining the concentration of lead in water under
the Safe Drinking Water Act, for example, the analyst must use a protocol
specified by the Environmental Protection Agency.
There is an obvious order to these four levels of analytical methodology.
Ideally, a protocol uses a previously validated procedure. Before developing
and validating a procedure, a method of analysis must be selected. This
requires, in turn, an initial screening of available techniques to determine
those that have the potential for monitoring the analyte.

3C Classifying Analytical Techniques

The analysis of a sample generates a chemical or physical signal that is
1 2
proportional to the amount of analyte in the sample. This signal may be
anything we can measure, such as volume or absorbance. It is convenient to
divide analytical techniques into two general classes based on whether the
signal is proportional to the mass or moles of analyte, or is proportional to
the analyte’s concentration.
Consider the two graduated cylinders in Figure 3.3, each of which
contains a solution of 0.010 M Cu(NO3)2. Cylinder 1 contains 10 mL, or
1.0 × 10-4 moles of Cu2+, and cylinder 2 contains 20 mL, or 2.0 × 10-4 moles
of Cu2+. If a technique responds to the absolute amount of analyte in the
sample, then the signal due to the analyte, SA, is
Figure 3.3 Two graduated cyl-
inders, each containing 0.10 M SA = kAnA 3.1
Cu(NO3)2. Although the cylin- where nA is the moles or grams of analyte in the sample, and kA is a pro-
ders contain the same concentra- portionality constant. Because cylinder 2 contains twice as many moles of
tion of Cu2+, the cylinder on the Cu2+ as cylinder 1, analyzing the contents of cylinder 2 gives a signal twice
left contains 1.0 × 10-4 mol Cu2+ as large that for cylinder 1.
and the cylinder on the right con- A second class of analytical techniques are those that respond to the
tains 2.0 × 10-4 mol Cu2+. analyte’s concentration, CA
SA = kACA 3.2
Since the solutions in both cylinders have the same concentration of Cu2+,
their analysis yields identical signals.
A technique that responds to the absolute amount of analyte is a total
analysis technique. Mass and volume are the most common signals for a
Historically, most early analytical meth- total analysis technique, and the corresponding techniques are gravimetry
ods used a total analysis technique. For
this reason, total analysis techniques are (Chapter 8) and titrimetry (Chapter 9). With a few exceptions, the signal
often called “classical” techniques. for a total analysis technique is the result of one or more chemical reactions,
the stoichiometry of which determines the value of kA in equation 3.1.
 Chapter 3 The Vocabulary of Analytical Chemistry 45

Spectroscopy (Chapter 10) and electrochemistry (Chapter 11), in which

an optical or an electrical signal is proportional to the relative amount of Since most concentration techniques rely
analyte in a sample, are examples of concentration techniques. The re- on measuring an optical or electrical sig-
nal, they also are known as “instrumental”
lationship between the signal and the analyte’s concentration is a theoretical
techniques.
function that depends on experimental conditions and the instrumentation
used to measure the signal. For this reason the value of kA in equation 3.2
is determined experimentally.

3D Selecting an Analytical Method

A method is the application of a technique to a specific analyte in a specific
matrix. We can develop an analytical method to determine the concentra-
tion of lead in drinking water using any of the techniques mentioned in
the previous section. A gravimetric method, for example, might precipi-
tate the lead as PbSO4 or as PbCrO4, and use the precipitate’s mass as the
analytical signal. Lead forms several soluble complexes, which we can use
to design a complexation titrimetric method. As shown in Figure 3.2, we
can use graphite furnace atomic absorption spectroscopy to determine the
concentration of lead in drinking water. Finally, lead’s multiple oxidation
states (Pb0, Pb2+, Pb4+) makes feasible a variety of electrochemical methods.
Ultimately, the requirements of the analysis determine the best method.
In choosing among the available methods, we give consideration to some
or all the following design criteria: accuracy, precision, sensitivity, selectiv-
ity, robustness, ruggedness, scale of operation, analysis time, availability of
equipment, and cost.

3D.1 Accuracy
Because it is unlikely that we know the
Accuracy is how closely the result of an experiment agrees with the “true” true result, we use an expected or accepted
result to evaluate accuracy. For example,
or expected result. We can express accuracy as an absolute error, e we might use a standard reference mate-
rial, which has an accepted value, to estab-
e = obtained result - expected result lish an analytical method’s accuracy.
or as a percentage relative error, %er
obtained result - expected result
%e r = # 100
expected result
A method’s accuracy depends on many things, including the signal’s source,
the value of kA in equation 3.1 or equation 3.2, and the ease of handling You will find a more detailed treatment of
samples without loss or contamination. A total analysis technique, such as accuracy in Chapter 4, including a discus-
gravimetry and titrimetry, often produce more accurate results than does sion of sources of errors.
a concentration technique because we can measure mass and volume with
high accuracy, and because the value of kA is known exactly through stoi-
chiometry.
46 Analytical Chemistry 2.1

(a)

5.8 5.9 6.0 6.1 6.2

ppm K

(b)

5.8 5.9 6.0 6.1 6.2

ppm K

Figure 3.4 Two determinations of the concentration of K+ in serum, showing the

effect of precision on the distribution of individual results. The data in (a) are less
scattered and, therefore, more precise than the data in (b).

3D.2 Precision
When a sample is analyzed several times, the individual results vary from
Confusing accuracy and precision is a
trial-to-trial. Precision is a measure of this variability. The closer the agree-
common mistake. See Ryder, J.; Clark, A. ment between individual analyses, the more precise the results. For example,
U. Chem. Ed. 2002, 6, 1–3, and Tomlin- the results shown in Figure 3.4(a) for the concentration of K+ in a sample
son, J.; Dyson, P. J.; Garratt, J. U. Chem.
Ed. 2001, 5, 16–23 for discussions of this
of serum are more precise than those in Figure 3.4(b). It is important to
and other common misconceptions about understand that precision does not imply accuracy. That the data in Figure
the meaning of error. 3.4(a) are more precise does not mean that the first set of results is more
accurate. In fact, neither set of results may be accurate.
A method’s precision depends on several factors, including the uncer-
tainty in measuring the signal and the ease of handling samples reproduc-
You will find a more detailed treatment of
precision in Chapter 4, including a discus-
ibly. In most cases we can measure the signal for a total analysis technique
sion of sources of errors. with a higher precision than is the case for a concentration method. Preci-
sion is covered in more detail in Chapter 4.

3D.3 Sensitivity

Confidence, as we will see in Chapter 4,

The ability to demonstrate that two samples have different amounts of ana-
is a statistical concept that builds on the lyte is an essential part of many analyses. A method’s sensitivity is a mea-
idea of a population of results. For this sure of its ability to establish that such a difference is significant. Sensitivity
reason, we will postpone our discussion of
detection limits to Chapter 4. For now,
is often confused with a method’s detection limit, which is the smallest
the definition of a detection limit given amount of analyte we can determine with confidence.
here is sufficient. Sensitivity is equivalent to the proportionality constant, kA, in equa-
tion 3.1 and equation 3.2.3 If DSA is the smallest difference we can measure

3 IUPAC Compendium of Chemical Terminology, Electronic version, http://goldbook.iupac.org/

S05606.html.
 Chapter 3 The Vocabulary of Analytical Chemistry 47

between two signals, then the smallest detectable difference in the absolute
amount or the relative amount of analyte is
3nA = 3SA or 3 C A = 3 S A
kA kA
Suppose, for example, that our analytical signal is a measurement of mass
using a balance whose smallest detectable increment is ±0.0001 g. If our
method’s sensitivity is 0.200, then our method can conceivably detect a
difference in mass of as little as
! 0.0001 g
3 n A = 0.200 = ! 0.0005 g
For two methods with the same DSA, the method with the greater sensitiv-
ity—that is, the method with the larger kA—is better able to discriminate
between smaller amounts of analyte.

3D.4 Specificity and Selectivity

An analytical method is specific if its signal depends only on the analyte.4
Although specificity is the ideal, few analytical methods are free from
interferences. When an interferent contributes to the signal, we expand
equation 3.1 and equation 3.2 to include its contribution to the sample’s
signal, Ssamp
S samp = S A + S I = k A n A + k I n I 3.3
S samp = S A + S I = k A C A + k I C I 3.4
where SI is the interferent’s contribution to the signal, kI is the interferent’s
sensitivity, and nI and CI are the moles (or grams) and the concentration of
interferent in the sample, respectively.
Selectivity is a measure of a method’s freedom from interferences.5 A
method’s selectivity for an interferent relative to the analyte is defined by a
selectivity coefficient, K­A,I

K A,I = k I 3.5
kA
Although kA and kI usually are positive,
which may be positive or negative depending on the sign of kI and kA. The they can be negative. For example, some
selectivity coefficient is greater than +1 or less than –1 when the method analytical methods work by measuring the
concentration of a species that remains
is more selective for the interferent than for the analyte. after is reacts with the analyte. As the
Determining the selectivity coefficient’s value is easy if we already know analyte’s concentration increases, the con-
the values for kA and kI. As shown by Example 3.1, we also can deter- centration of the species that produces the
signal decreases, and the signal becomes
mine KA,I by measuring Ssamp in the presence of and in the absence of the smaller. If the signal in the absence of ana-
interferent. lyte is assigned a value of zero, then the
subsequent signals are negative.

4 (a) Persson, B-A; Vessman, J. Trends Anal. Chem. 1998, 17, 117–119; (b) Persson, B-A; Vessman,
J. Trends Anal. Chem. 2001, 20, 526–532.
5 Valcárcel, M.; Gomez-Hens, A.; Rubio, S. Trends Anal. Chem. 2001, 20, 386–393.
48 Analytical Chemistry 2.1

Example 3.1
A method for the analysis of Ca2+ in water suffers from an interference in
the presence of Zn2+. When the concentration of Ca2+ is 100 times greater
than that of Zn2+, an analysis for Ca2+ has a relative error of +0.5%. What
is the selectivity coefficient for this method?
Solution
Since only relative concentrations are reported, we can arbitrarily assign ab-
solute concentrations. To make the calculations easy, we will let CCa = 100
(arbitrary units) and CZn = 1. A relative error of +0.5% means the signal in
If you are unsure why the signal in the
presence of zinc is 100.5, note that the the presence of Zn2+ is 0.5% greater than the signal in the absence of Zn2+.
percentage relative error for this problem Again, we can assign values to make the calculation easier. If the signal for
is given by Cu2+ in the absence of Zn2+ is 100 (arbitrary units), then the signal in the
obtained result - 100 presence of Zn2+ is 100.5.
#
100
The value of kCa is determined using equation 3.2
100 = + 0.5%
Solving gives an obtained result of 100.5. S Ca = 100 = 1
k Ca = C
Ca 100
In the presence of Zn2+ the signal is given by equation 3.4; thus
S samp = 100.5 = k Ca C Ca + k Zn C Zn = (1 # 100) + k Zn # 1
Solving for kZn gives its value as 0.5. The selectivity coefficient is
K Ca,Zn = k Zn = 01.5 = 0.5
k Ca

Practice Exercise 3.1

Wang and colleagues describe a fluorescence method for the analysis of
Ag+ in water. When analyzing a solution that contains 1.0 × 10-9 M Ag+
and 1.1× 10-7 M Ni2+, the fluorescence intensity (the signal) was +4.9%
greater than that obtained for a sample of 1.0 × 10-9 M Ag+. What is
KAg,Ni for this analytical method? The full citation for the data in this
exercise is Wang, L.; Liang, A. N.; Chen, H.; Liu, Y.; Qian, B.; Fu, J.
Anal. Chim. Acta 2008, 616, 170-176.
Click here to review your answer to this exercise.

A selectivity coefficient provides us with a useful way to evaluate an

interferent’s potential effect on an analysis. Solving equation 3.5 for kI
k I = K A,I # k A 3.6
substituting in equation 3.3 and equation 3.4, and simplifying gives
S samp = k A {n A + K A,I # n A} 3.7
S samp = k A {C A + K A,I # C I } 3.8
 Chapter 3 The Vocabulary of Analytical Chemistry 49

An interferent will not pose a problem as long as the term KA,I × nI in equa-
tion 3.7 is significantly smaller than nA, or if KA,I × CI in equation 3.8 is
significantly smaller than CA.

Example 3.2
Barnett and colleagues developed a method to determine the concentra-
tion of codeine in poppy plants.6 As part of their study they evaluated the
effect of several interferents. For example, the authors found that equimo- H3CO
lar solutions of codeine and the interferent 6-methoxycodeine gave signals,
respectively of 40 and 6 (arbitrary units).
(a) What is the selectivity coefficient for the interferent, 6-methoxyco-
O H
deine, relative to that for the analyte, codeine.
(b) If we need to know the concentration of codeine with an accuracy of H N
±0.50%, what is the maximum relative concentration of 6-methoxy- CH3

codeine that we can tolerate? HO

Solution codeine

(a) The signals due to the analyte, SA, and the interferent, SI, are
SA = kACA SI = kI CI
Solving these equations for kA and for kI, and substituting into equa-
tion 3.6 gives
K A,I = SS I //C
CI
A A

Because the concentrations of analyte and interferent are equimolar

(CA = CI), the selectivity coefficient is
K A,I = SS I = 6 = 0.15
A 40
(b) To achieve an accuracy of better than ±0.50% the term KA,I × CI in
equation 3.8 must be less than 0.50% of CA; thus
K A,I # C I # 0.0050 # C A
Solving this inequality for the ratio CI/CA and substituting in the
value for KA,I from part (a) gives
CI 0.0050 0.0050
C A # K A,I = 0.15 = 0.033
Therefore, the concentration of 6-methoxycodeine must be less than
3.3% of codeine’s concentration.

When a method’s signal is the result of a chemical reaction—for exam-

ple, when the signal is the mass of a precipitate—there is a good chance that
the method is not very selective and that it is susceptible to an interference.
6 Barnett, N. W.; Bowser, T. A.; Geraldi, R. D.; Smith, B. Anal. Chim. Acta 1996, 318, 309–
317.
50 Analytical Chemistry 2.1

Practice Exercise 3.2

Mercury (II) also is an interferent in the fluorescence method for Ag+
developed by Wang and colleagues (see Practice Exercise 3.1 for the cita-
tion). The selectivity coefficient, KAg,Hg has a value of –1.0 × 10–3.
(a) What is the significance of the selectivity coefficient’s negative sign?
(b) Suppose you plan to use this method to analyze solutions with con-
centrations of Ag+ no smaller than 1.0 nM . What is the maximum
concentration of Hg2+ you can tolerate if your percentage relative
errors must be less than ±1.0%?
Look back at Figure 1.1, which shows Fre-
senius’ analytical method for the determi- Click here to review your answers to this exercise.
nation of nickel in ores. The reason there
are so many steps in this procedure is that Problems with selectivity also are more likely when the analyte is present at
precipitation reactions generally are not
very selective. The method in Figure 1.2
a very low concentration.7
includes fewer steps because dimethylgly-
oxime is a more selective reagent. Even so, 3D.5 Robustness and Ruggedness
if an ore contains palladium, additional
steps are needed to prevent the palladium For a method to be useful it must provide reliable results. Unfortunately,
from interfering. methods are subject to a variety of chemical and physical interferences that
contribute uncertainty to the analysis. If a method is relatively free from
chemical interferences, we can use it to analyze an analyte in a wide variety
of sample matrices. Such methods are considered robust.
Random variations in experimental conditions introduces uncertainty.
If a method’s sensitivity, k, is too dependent on experimental conditions,
such as temperature, acidity, or reaction time, then a slight change in any of
these conditions may give a significantly different result. A rugged method
is relatively insensitive to changes in experimental conditions.

3D.6 Scale of Operation

Another way to narrow the choice of methods is to consider three potential
limitations: the amount of sample available for the analysis, the expected
concentration of analyte in the samples, and the minimum amount of ana-
lyte that will produce a measurable signal. Collectively, these limitations
define the analytical method’s scale of operations.
We can display the scale of operations visually (Figure 3.5) by plot-
ting the sample’s size on the x-axis and the analyte’s concentration on the
y-axis.8 For convenience, we divide samples into macro (>0.1 g), meso (10
mg–100 mg), micro (0.1 mg–10 mg), and ultramicro (<0.1 mg) sizes, and
we divide analytes into major (>1% w/w), minor (0.01% w/w–1% w/w),
trace (10-7% w/w–0.01% w/w), and ultratrace (<10–7% w/w) components.
Together, the analyte’s concentration and the sample’s size provide a charac-
teristic description for an analysis. For example, in a microtrace analysis the
7 Rodgers, L. B. J. Chem. Educ. 1986, 63, 3–6.
8 (a) Sandell, E. B.; Elving, P. J. in Kolthoff, I. M.; Elving, P. J., eds. Treatise on Analytical Chem-
istry, Interscience: New York, Part I, Vol. 1, Chapter 1, pp. 3–6; (b) Potts, L. W. Quantitative
Analysis–Theory and Practice, Harper and Row: New York, 1987, pp. 12.
 Chapter 3 The Vocabulary of Analytical Chemistry 51

sample weighs between 0.1 mg and 10 mg and contains a concentration of

analyte between 10–7% w/w and 10–2% w/w.
The diagonal lines connecting the axes show combinations of sample
size and analyte concentration that contain the same absolute mass of ana-
lyte. As shown in Figure 3.5, for example, a 1-g sample that is 1% w/w
analyte has the same amount of analyte (10 mg) as a 100-mg sample that
is 10% w/w analyte, or a 10-mg sample that is 100% w/w analyte.
We can use Figure 3.5 to establish limits for analytical methods. If a
method’s minimum detectable signal is equivalent to 10 mg of analyte, then
it is best suited to a major analyte in a macro or meso sample. Extending the
method to an analyte with a concentration of 0.1% w/w requires a sample
of 10 g, which rarely is practical due to the complications of carrying such It should not surprise you to learn that a
a large amount of material through the analysis. On the other hand, a small total analysis technique typically requires
sample that contains a trace amount of analyte places significant restric- a macro or a meso sample that contains a
major analyte. A concentration technique
tions on an analysis. For example, a 1-mg sample that is 10–4% w/w in is particularly useful for a minor, trace,
analyte contains just 1 ng of analyte. If we isolate the analyte in 1 mL of or ultratrace analyte in a macro, meso, or
solution, then we need an analytical method that reliably can detect it at a micro sample.
concentration of 1 ng/mL.

10–9 %

ultratrace 10–8 %

10–7 % ppb
concentration of analyte (% w/w)

10–6 %

10–5 % microtrace
trace analysis
10–4 % ppm

10–3 %

10–2 %
Figure 3.5 Scale of operations for ana-
minor 10–1 % lytical methods (adapted from refer-
ences 8a and 8b).
1%
The shaded areas define different types
major 10 % 10 of analyses. The boxed area, for exam-
m
10

g ple, represents a microtrace analysis.

1
0

μg
μg

100 % The diagonal lines show combinations

1 10–1 10–2 10–3 10–4 10–5 10–6 10–7 10–8 10–9 of sample size and analyte concentra-
1 mg 1μg 1 ng tion that contain the same mass of
macro meso micro ultramicro •
analyte. The three filled circles ( ), for
example, indicate analyses that use 10
mass of sample (g) mg of analyte.
52 Analytical Chemistry 2.1

3D.7 Equipment, Time, and Cost

Finally, we can compare analytical methods with respect to their equip-
ment needs, the time needed to complete an analysis, and the cost per
sample. Methods that rely on instrumentation are equipment-intensive
and may require significant operator training. For example, the graphite
furnace atomic absorption spectroscopic method for determining lead in
water requires a significant capital investment in the instrument and an
experienced operator to obtain reliable results. Other methods, such as
titrimetry, require less expensive equipment and less training.
The time to complete an analysis for one sample often is fairly similar
from method-to-method. This is somewhat misleading, however, because
much of this time is spent preparing samples, preparing reagents, and gath-
ering together equipment. Once the samples, reagents, and equipment are
in place, the sampling rate may differ substantially. For example, it takes
just a few minutes to analyze a single sample for lead using graphite fur-
nace atomic absorption spectroscopy, but several hours to analyze the same
sample using gravimetry. This is a significant factor in selecting a method
for a laboratory that handles a high volume of samples.
The cost of an analysis depends on many factors, including the cost of
equipment and reagents, the cost of hiring analysts, and the number of
samples that can be processed per hour. In general, methods that rely on
instruments cost more per sample then other methods.

3D.8 Making the Final Choice

Unfortunately, the design criteria discussed in this section are not mutually
independent.9 Working with smaller samples or improving selectivity often
comes at the expense of precision. Minimizing cost and analysis time may
decrease accuracy. Selecting a method requires carefully balancing the vari-
ous design criteria. Usually, the most important design criterion is accuracy,
and the best method is the one that gives the most accurate result. When
the need for a result is urgent, as is often the case in clinical labs, analysis
time may become the critical factor.
In some cases it is the sample’s properties that determine the best meth-
od. A sample with a complex matrix, for example, may require a method
with excellent selectivity to avoid interferences. Samples in which the ana-
lyte is present at a trace or ultratrace concentration usually require a con-
centration method. If the quantity of sample is limited, then the method
must not require a large amount of sample.
Determining the concentration of lead in drinking water requires a
method that can detect lead at the parts per billion concentration level.
Selectivity is important because other metal ions are present at significantly
higher concentrations. A method that uses graphite furnace atomic absorp-
tion spectroscopy is a common choice for determining lead in drinking
9 Valcárcel, M.; Ríos, A. Anal. Chem. 1993, 65, 781A–787A.
 Chapter 3 The Vocabulary of Analytical Chemistry 53

water because it meets these specifications. The same method is also useful
for determining lead in blood where its ability to detect low concentrations
of lead using a few microliters of sample is an important consideration.

3E Developing the Procedure

After selecting a method, the next step is to develop a procedure that accom-
plish our goals for the analysis. In developing a procedure we give attention
to compensating for interferences, to selecting and calibrating equipment,
to acquiring a representative sample, and to validating the method.

3E.1 Compensating for Interferences

A method’s accuracy depends on its selectivity for the analyte. Even the best
method, however, may not be free from interferents that contribute to the
measured signal. Potential interferents may be present in the sample itself
or in the reagents used during the analysis.
When the sample is free of interferents, the total signal, Stotal, is a sum
of the signal due to the analyte, SA, and the signal due to interferents in
the reagents, Sreag,
S total = S A + S reag = k A n A + S reag 3.9
S total = S A + S reag = k A C A + S reag 3.10
Without an independent determination of Sreag we cannot solve equation
3.9 or 3.10 for the moles or concentration of analyte.
To determine the contribution of Sreag in equations 3.9 and 3.10 we
measure the signal for a method blank, a solution that does not contain
the sample. Consider, for example, a procedure in which we dissolve a 0.1-g
sample in a portion of solvent, add several reagents, and dilute to 100 mL
with additional solvent. To prepare the method blank we omit the sample
and dilute the reagents to 100 mL using the solvent. Because the analyte is A method blank also is known as a reagent
absent, Stotal for the method blank is equal to Sreag. Knowing the value for blank.
Sreag makes it is easy to correct Stotal for the reagent’s contribution to the
total signal; thus When the sample is a liquid, or is in so-
lution, we use an equivalent volume of
(S total - S reag ) = S A = k A n A an inert solvent as a substitute for the
sample.
(S total - S reag ) = S A = k A C A

By itself, a method blank cannot compensate for an interferent that is

part of the sample’s matrix. If we happen to know the interferent’s identity
and concentration, then we can be add it to the method blank; however,
this is not a common circumstance and we must, instead, find a method
for separating the analyte and interferent before continuing the analysis.
54 Analytical Chemistry 2.1

Methods for effecting this separation are 3E.2 Calibration

discussed in Chapter 7.
A simple definition of a quantitative analytical method is that it is a mecha-
nism for converting a measurement, the signal, into the amount of analyte
in a sample. Assuming we can correct for interferents, a quantitative analysis
is nothing more than solving equation 3.1 or equation 3.2 for nA or for CA.
To solve these equations we need the value of kA. For a total analysis
method usually we know the value of kA because it is defined by the stoi-
chiometry of the chemical reactions responsible for the signal. For a con-
centration method, however, the value of kA usually is a complex function
of experimental conditions. A Calibration is the process of experimentally
determining the value of kA by measuring the signal for one or more stan-
dard samples, each of which contains a known concentration of analyte.
With a single standard we can calculate the value of kA using equation 3.1
or equation 3.2. When using several standards with different concentra-
tions of analyte, the result is best viewed visually by plotting SA versus the
concentration of analyte in the standards. Such a plot is known as a cali-
bration curve, an example of which is shown in Figure 3.6.

3E.3 Sampling
Selecting an appropriate method and executing it properly helps us ensure
that our analysis is accurate. If we analyze the wrong sample, however, then
the accuracy of our work is of little consequence.
A proper sampling strategy ensures that our samples are representative
of the material from which they are taken. Biased or nonrepresentative sam-
Chapter 7 provides a more detailed discus-
pling, and contaminating samples during or after their collection are two
sion of sampling, including strategies for
obtaining representative samples. examples of sampling errors that can lead to a significant error in accuracy.
It is important to realize that sampling errors are independent of errors in
the analytical method. As a result, we cannot correct a sampling error in
the laboratory by, for example, evaluating a reagent blank.

1
Figure 3.6 Example of a calibra-
•
tion curve. The filled circles ( ) are
0.8
the results for five standard sam-
ples, each with a different concen-
trations of analyte, and the line is 0.6
the best fit to the data determined
SA

by a linear regression analysis. See 0.4

Chapter 5 for a further discussion
of calibration curves and an expla-
0.2
nation of linear regression.

0
0.00 0.20 0.40 0.60 0.80 1.00
Concentration of Analyte (µg/mL)
 Chapter 3 The Vocabulary of Analytical Chemistry 55

3E.4 Validation
If we are to have confidence in our procedure we must demonstrate that it
can provide acceptable results, a process we call validation. Perhaps the
most important part of validating a procedure is establishing that its preci-
sion and accuracy are appropriate for the problem we are trying to solve. We
also ensure that the written procedure has sufficient detail so that different
analysts or laboratories will obtain comparable results. Ideally, validation
uses a standard sample whose composition closely matches the samples we You will find more details about validating
analytical methods in Chapter 14.
will analyze. In the absence of appropriate standards, we can evaluate ac-
curacy by comparing results to those obtained using a method of known
accuracy.

3F Protocols
Earlier we defined a protocol as a set of stringent written guidelines that
specify an exact procedure that we must follow if an agency is to accept the
results of our analysis. In addition to the considerations that went into the
procedure’s design, a protocol also contains explicit instructions regarding
internal and external quality assurance and quality control (QA/QC) proce-
dures.10 The goal of internal QA/QC is to ensure that a laboratory’s work
is both accurate and precise. External QA/QC is a process in which an
external agency certifies a laboratory.
As an example, let’s outline a portion of the Environmental Protection
Agency’s protocol for determining trace metals in water by graphite furnace
atomic absorption spectroscopy as part of its Contract Laboratory Program
(CLP). The CLP protocol (see Figure 3.7) calls for an initial calibration
using a method blank and three standards, one of which is at the detec-
tion limit. The resulting calibration curve is verified by analyzing initial
calibration verification (ICV) and initial calibration blank (ICB) samples.
The lab’s result for the ICV sample must fall within ±10% of its expected
concentration. If the result is outside this limit the analysis is stopped and
the problem identified and corrected before continuing.
After a successful analysis of the ICV and ICB samples, the lab reverifies
the calibration by analyzing a continuing calibration verification (CCV)
sample and a continuing calibration blank (CCB). Results for the CCV also
must be within ±10% of its expected concentration. Again, if the lab’s result
for the CCV is outside the established limits, the analysis is stopped, the
problem identified and corrected, and the system recalibrated as described
above. Additional CCV and the CCB samples are analyzed before the first
sample and after the last sample, and between every set of ten samples. If
the result for any CCV or CCB sample is unacceptable, the results for the
last set of samples are discarded, the system is recalibrated, and the samples
reanalyzed. By following this protocol, each result is bound by successful
10 (a) Amore, F. Anal. Chem. 1979, 51, 1105A–1110A; (b) Taylor, J. K. Anal. Chem. 1981, 53,
1588A–1593A.
56 Analytical Chemistry 2.1

Start

Figure 3.7 Schematic diagram showing a portion of the EPA’s

protocol for determining trace metals in water using graphite Initial Calibration
furnace atomic absorption spectrometry.
The abbreviations are ICV: initial calibration verification;
ICB: initial calibration blank; CCV: continuing calibration
ICV, ICB No Identify and
verification; CCB: continuing calibration blank. correct problem
OK?

Yes

CCV, CCB No
OK?

Yes

Run 10 Samples

CCV, CCB No Discard results for

OK? last set of samples

Yes

Yes More
Samples?

No

End

checks on the calibration. Although not shown in Figure 3.7, the protocol
also contains instructions for analyzing duplicate or split samples, and for
using spike tests to verify accuracy.

3G The Importance of Analytical Methodology

The importance of the issues raised in this chapter is evident if we examine
environmental monitoring programs. The purpose of a monitoring pro-
gram is to determine the present status of an environmental system, and to
assess long term trends in the system’s health. These are broad and poorly
defined goals. In many cases, an environmental monitoring program begins
before the essential questions are known. This is not surprising since it is
difficult to formulate questions in the absence of results. Without careful
planning, however, a poor experimental design may result in data that has
little value.
 Chapter 3 The Vocabulary of Analytical Chemistry 57

These concerns are illustrated by the Chesapeake Bay Monitoring Pro-

gram. This research program, designed to study nutrients and toxic pollut-
ants in the Chesapeake Bay, was initiated in 1984 as a cooperative venture
between the federal government, the state governments of Maryland, Vir-
ginia, and Pennsylvania, and the District of Columbia. A 1989 review of the
program highlights the problems common to many monitoring programs.11
At the beginning of the Chesapeake Bay monitoring program, little at-
tention was given to selecting analytical methods, in large part because the
eventual use of the data was not yet specified. The analytical methods ini-
tially chosen were standard methods already approved by the Environmen-
tal Protection Agency (EPA). In many cases these methods were not useful
because they were designed to detect pollutants at their legally mandated
maximum allowed concentrations. In unpolluted waters, however, the con-
centrations of these contaminants often are well below the detection limit
of the EPA methods. For example, the detection limit for the EPA approved
standard method for phosphate was 7.5 ppb. Since the actual phosphate
concentrations in Chesapeake Bay were below the EPA method’s detection
limit, it provided no useful information. On the other hand, the detection
limit for a non-approved variant of the EPA method, a method routinely
used by chemical oceanographers, was 0.06 ppb, a more realistic detec-
tion limit for their samples. In other cases, such as the elemental analysis
for particulate forms of carbon, nitrogen and phosphorous, EPA approved
procedures provided poorer reproducibility than nonapproved methods.

3H Key Terms
accuracy analysis analyte
calibration calibration curve concentration techniques
detection limit determination interferent
matrix measurement method
method blank precision procedure
protocol QA/QC robust
rugged selectivity selectivity coefficient
sensitivity signal specificity
technique total analysis techniques validation

3I Chapter Summary
Every discipline has its own vocabulary and your success in studying ana-
lytical chemistry will improve if you master this vocabulary. Be sure you
understand the difference between an analyte and its matrix, between a
technique and a method, between a procedure and a protocol, and between
a total analysis technique and a concentration technique.
11 D’Elia, C. F.; Sanders, J. G.; Capone, D. G. Envrion. Sci. Technol. 1989, 23, 768–774.
58 Analytical Chemistry 2.1

In selecting an analytical method we consider criteria such as accu-

racy, precision, sensitivity, selectivity, robustness, ruggedness, the amount
of available sample, the amount of analyte in the sample, time, cost, and
the availability of equipment. These criteria are not mutually independent,
and often it is necessary to find an acceptable balance between them.
In developing a procedure or protocol, we give consideration to com-
pensating for interferences, calibrating the method, obtaining an appropri-
ate sample, and validating the analysis. Poorly designed procedures and
protocols produce results that are insufficient to meet the needs of the
analysis.

3J Problems
1. When working with a solid sample, often it is necessary to bring the
analyte into solution by digesting the sample with a suitable solvent.
Any remaining solid impurities are removed by filtration before con-
tinuing with the analysis. In a typical total analysis method, the proce-
dure might read
After digesting the sample in a beaker using approximately 25 mL
of solvent, remove any solid impurities that remain by passing the
solution the analyte through filter paper, collecting the filtrate in a
clean Erlenmeyer flask. Rinse the beaker with several small portions
of solvent, passing these rinsings through the filter paper and col-
lecting them in the same Erlenmeyer flask. Finally, rinse the filter
paper with several portions of solvent, collecting the rinsings in the
same Erlenmeyer flask.

For a typical concentration method, however, the procedure might

state
After digesting the sample in a beaker using 25.00 mL of solvent,
remove any solid impurities by filtering a portion of the solution
containing the analyte. Collect and discard the first several mL of
filtrate before collecting a sample of 5.00 mL for further analysis.

Explain why these two procedures are different.

2. A certain concentration method works best when the analyte’s concen-

tration is approximately 10 ppb.
(a) If the method requires a sample of 0.5 mL, about what mass of
analyte is being measured?
(b) If the analyte is present at 10% w/v, how would you prepare the
sample for analysis?
(c) Repeat for the case where the analyte is present at 10% w/w.
 Chapter 3 The Vocabulary of Analytical Chemistry 59

(d) Based on your answers to parts (a)–(c), comment on the method’s

suitability for the determination of a major analyte.

3. An analyst needs to evaluate the potential effect of an interferent, I, on

the quantitative analysis for an analyte, A. She begins by measuring the
signal for a sample in which the interferent is absent and the analyte is
present with a concentration of 15 ppm, obtaining an average signal of
23.3 (arbitrary units). When she analyzes a sample in which the analyte
is absent and the interferent is present with a concentration of 25 ppm,
she obtains an average signal of 13.7.
(a) What is the sensitivity for the analyte?
(b) What is the sensitivity for the interferent?
(c) What is the value of the selectivity coefficient?
(d) Is the method more selective for the analyte or the interferent?
(e) What is the maximum concentration of interferent relative to that
of the analyte if the error in the analysis is to be less than 1%?

4. A sample is analyzed to determine the concentration of an analyte. Un-

der the conditions of the analysis the sensitivity is 17.2 ppm–1. What is
the analyte’s concentration if Stotal is 35.2 and Sreag is 0.6?

5. A method for the analysis of Ca2+ in water suffers from an interference

in the presence of Zn2+. When the concentration of Ca2+ is 50 times
greater than that of Zn2+, an analysis for Ca2+ gives a relative error of
–2.0%. What is the value of the selectivity coefficient for this method?

6. The quantitative analysis for reduced glutathione in blood is compli-

cated by many potential interferents. In one study, when analyzing a
solution of 10.0 ppb glutathione and 1.5 ppb ascorbic acid, the signal
was 5.43 times greater than that obtained for the analysis of 10.0 ppb
glutathione.12 What is the selectivity coefficient for this analysis? The
same study found that analyzing a solution of 3.5×102 ppb methio-
nine and 10.0 ppb glutathione gives a signal that is 0.906 times less
than that obtained for the analysis of 10.0 ppb glutathione. What is
the selectivity coefficient for this analysis? In what ways do these inter-
ferents behave differently?

7. Oungpipat and Alexander described a method for determining the con-

centration of glycolic acid (GA) in a variety of samples, including physi-
ological fluids such as urine.13 In the presence of only GA, the signal is

12 Jiménez-Prieto, R.; Velasco, A.; Silva, M; Pérez-Bendito, D. Anal. Chem. Acta 1992, 269, 273–
279.
13 Oungpipat, W.; Alexander, P. W. Anal. Chim. Acta 1994, 295, 36–46.
60 Analytical Chemistry 2.1

Ssamp,1 = kGACGA

and in the presence of both glycolic acid and ascorbic acid (AA), the
signal is

Ssamp,2 = kGACGA + kAACAA

When the concentration of glycolic acid is 1.0 × 10–4 M and the con-
centration of ascorbic acid is 1.0 × 10–5 M, the ratio of their signals is
S samp,1
S samp,2 = 1.44
(a) Using the ratio of the two signals, determine the value of the selec-
tivity ratio KGA,AA.
(b) Is the method more selective toward glycolic acid or ascorbic acid?
(c) If the concentration of ascorbic acid is 1.0 × 10–5 M, what is the
smallest concentration of glycolic acid that can be determined such
that the error introduced by failing to account for the signal from
ascorbic acid is less than 1%?

8. Ibrahim and co-workers developed a new method for the quantitative

analysis of hypoxanthine, a natural compound of some nucleic acids.14
As part of their study they evaluated the method’s selectivity for hy-
poxanthine in the presence of several possible interferents, including
ascorbic acid.
(a) When analyzing a solution of 1.12 × 10–6 M hypoxanthine the au-
thors obtained a signal of 7.45 × 10–5 amps. What is the sensitivity
for hypoxanthine? You may assume the signal has been corrected
for the method blank.
(b) When a solution containing 1.12 × 10–6 M hypoxanthine and
6.5 × 10–5 M ascorbic acid is analyzed a signal of 4.04 × 10–5 amps
is obtained. What is the selectivity coefficient for this method?
(c) Is the method more selective for hypoxanthine or for ascorbic
acid?
(d) What is the largest concentration of ascorbic acid that may be pres-
ent if a concentration of 1.12 × 10–6 M hypoxanthine is to be de-
termined within 1.0%?

9. Examine a procedure from Standard Methods for the Analysis of Waters

and Wastewaters (or another manual of standard analytical methods)
and identify the steps taken to compensate for interferences, to cali-

14 Ibrahim, M. S.; Ahmad, M. E.; Temerk, Y. M.; Kaucke, A. M. Anal. Chim. Acta 1996, 328,
47–52.
 Chapter 3 The Vocabulary of Analytical Chemistry 61

brate equipment and instruments, to standardize the method, and to

acquire a representative sample.

3K Solutions to Practice Exercises

Practice Exercise 3.1
Because the signal for Ag+ in the presence of Ni2+ is reported as a relative
error, we will assign a value of 100 as the signal for 1 × 10–9 M Ag+. With a
relative error of +4.9%, the signal for the solution of 1 × 10–9 M Ag+ and
1.1 × 10–7 M Ni2+ is 104.9. The sensitivity for Ag+ is determined using the
solution that does not contain Ni2+; thus
S Ag 100
k Ag = C = = 1.0 # 10 11 M -1
Ag 1 # 10 -9 M
Substituting into equation 3.4 values for kAg, Ssamp , and the concentrations
of Ag+ and Ni2+
104.9 = (1.0 # 10 11 M -1) # (1.0 # 10 -9 M) + k Ni # (1.1 # 10 -7 M)
and solving gives kNi as 4.5 × 107 M–1. The selectivity coefficient is

K Ag,Ni = k Ni = 4.5 # 1011 M -1 = 4.5 # 10 -4

7 -1

k Ag 1.0 # 10 M
Click here to return to the chapter.

Practice Exercise 3.2

(a) A negative value for KAg,Hg means that the presence of Hg2+ decreases
the signal from Ag+.
(b) In this case we need to consider an error of –1%, since the effect of Hg2+
is to decrease the signal from Ag+. To achieve this error, the term KA,I × CI
in equation 3.8 must be less than -1% of CA; thus
K Ag,Hg # C Hg =- 0.01 # C Ag
Substituting in known values for KAg,Hg and CAg, we find that the maxi-
mum concentration of Hg2+ is 1.0 × 10-8 M.
Click here to return to the chapter.
62 Analytical Chemistry 2.1