Trends in Analytical Chemistry, Vol. 24, No.

6, 2005 Trends

Quality control in qualitative

analysis
B.M. Simonet

In recent years, quality control (QC) has acquired great importance in vide trust about meeting the quality
chemical laboratories, so the international standard ISO/IEC 17025 points requirements [2]. In a more specific
out that any laboratory shall establish, implement and maintain a quality approach, international standard ISO/IEC
system appropriate to the scope of its activity. However, the standard 17025 points out that any laboratory
scarcely addresses qualitative analysis in spite of its increasing importance in shall establish, implement and maintain a
chemical and biochemical laboratories. In part of the report EUR 20605 EN quality system appropriate to the scope of
(2002), QC of qualitative analysis was considered for the first time. This its activity [3]. In other words, the labo-
overview presents one view of the state of the art of QC in qualitative ratory shall have QC procedures for mon-
analysis. The use of control charts in qualitative analysis is the most impor- itoring the validity of test and calibrations
tant activity of internal QC to monitor and control routine analysis. The undertaken.
paper describes the control charts to be used in qualitative analysis because In general, quality can be defined as
the control charts typically used in quantitative analysis cannot be used the degree of fulfilment of expectations
directly. The correct application of proficiency testing (PT) schemes to express, as well as implied. Depending on
qualitative analysis, the most important activity of external QC, is also these expectations, different types of
described in this overview. quality can be defined. The term QA
ª 2005 Elsevier Ltd. All rights reserved. describes the overall measures that a
laboratory uses to ensure the quality of
Keywords: Control chart; EQC; External quality control; Internal quality control; IQC;
its operations [4]. It is important to
Proficiency testing scheme; PT; QC; Qualitative analysis; Quality control
distinguish between QA and QC. In fact
the meaning of these terms often varies
1. Introduction according to the context. But, in practical
B.M. Simonet*
terms, we can consider, as was indicated
Department of Analytical
Chemistry, University of Chemical analysis or measurement gen- above, that QA relates to the overall
Girona, Campus de Montilivi, erally involves the use of a validated measures taken by the laboratory to
E-17071 Girona, Spain method, the use of an adequate calibration regulate or ensure quality, whereas QC
graph to ensure traceability and the describes the individual measures that
measurement of uncertainty. These relate to the quality of individual samples
actions form the first level of quality or batches of samples. Typical QC proce-
assurance (QA) in the laboratory. But, dures include: (i) analysis of reference
only when well-defined quality control materials/measurement standards; (ii)
(QC) procedures are also employed the analysis of blind samples; (iii) use of QC
validity of measurement can be assured samples and control charts; (iv) analysis
(Fig. 1). In fact, QC and quality in chemi- of blanks; (v) analysis of spiked samples;
cal analysis are old concepts. Probably, as (vi) analysis in duplicate; and, (vii)
long as there has been chemical analysis, proficiency testing (PT).
there has been the need to control the The CITAC/Eurachem guide to quality
quality of its performance [1]. However, in analytical chemistry points out that, as
due to the enforcement of legal require- a part of their quality systems and to
ments and the high standards between monitor day-to-day and batch-to-batch
companies obliged to assure the quality of analytical performance, laboratories must
the results, QC seems a young concept. operate an appropriate level of internal QC
*
Fax: +34 971 418150;
The new international standard ISO checks and participate wherever possible
E-mail: 9000:2000 defines QA as a part of quality in appropriate PT schemes (external QC)
bartolome.simonet@udg.es. management addressing the need to pro- [5]. Obviously, the level and the type of QC

0165-9936/$ - see front matter ª 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.trac.2005.03.011 525
Trends Trends in Analytical Chemistry, Vol. 24, No. 6, 2005

2. QC in qualitative analysis
Validation

To control the quality of results, it is necessary to

Quality Control
Valid and Safe
Meas urement
Traceable Measurement
Response/ Calibration Uncertainty/Traceability
Figure 1. Role of quality control in the analytical process to obtain
valid and safe measurements.
will critically depend on the nature of the analysis, the
frequency of the analysis, the batch size, the degree of
automation, and the test difficulty and reliability.
The importance of establishing the QC principles is
clear. They have been established by the international
standard ISO/IEC 17025. Unfortunately, this inter-
national standard scarcely addresses qualitative analysis
in spite of the fact that qualitative analyses represent a
significant and important part of the daily activity of
chemical and biochemical laboratories. Such importance
was reported in EUR 18405 EN (1998) [6]. Then, EUR
20605 EN (2002) systematically considered the
metrology of qualitative chemical analysis. An important
part of EUR 20605 EN deals with the QC of qualitative
analysis [7,8].
This article presents an overview of the state of the art
of QC in qualitative analysis.
describe the parameter that denotes the quality of a
certain property of chemical analysis. Afterwards, it is
necessary to define how the quality can be influenced
and what features determine the optimal quality.
Quality is always a relative notion, referring to the
requirements fixed beforehand on the basis of national or
international regulations or customer needs [9]. In this
context, the requirements fixed by a screening or quali-
tative method are clear, because, as is well known, the
goal of a screening method based on a binary response is
the classification of samples according to the presence/
absence of an analyte or a group of analytes. The QC for
this type of method therefore focuses on identifying false
positives and false negatives.
QC can be classified into two groups: internal QC (IQC)
and external QC (EQC). Fig. 2 describes the types of QC
and the activities carried out in each case.
IQC has, as its main objective, the maintenance of
validation conditions in the laboratory for a long time. It
is meant to assure the appropriate performance of the
particular test used for qualitative analysis. Another key
principle for quality is comparability of results between
laboratories, which is precisely the objective of EQC – to
ensure that qualitative analysis performs adequately in
an interlaboratory test.
It is important to note that data about the quality of
analytical measurements encompasses two essential
criteria: utility and reliability. Utility means that ana-
lytical results must allow reliable decision making (e.g., if
a result is a false positive, the decision will be wrong).

QUALITY CONTOL
(QC)

Internal Quality Control External Quality Control

(IQC) (EQC)

OBJECTIVE To maintain the validation OBJECTIVE To assure the

conditions in the comparability of results
laboratory between laboratories

ACTIVITIES ACTIVITIES
Repetition of analysis Proficiency testing
Comparison between schemes
analysts
Use of reference materials
Control charts
Etc.

Figure 2. Classification of quality control according to its objective and activities.

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Trends in Analytical Chemistry, Vol. 24, No. 6, 2005
The consequences of a wrong decision can be health
risks, higher costs or even illegal practices. IQC and EQC
are therefore key tools to assure quality.
3. (Certified) reference materials
Reference materials (RMs) are important tools in realis-
ing a number of aspects of measurement quality
because, in addition to their use for method validation,
calibration or estimation of measurement uncertainty,
they are also used for IQC and EQC. Matrix RMs, which
are characterised for the composition of specific major or
minor constituents, are the RMs most widely used for
QC.
In general, RMs can be classified into two classes
according to ISO recommendations: (i) certified RMs
(CRMs) and (ii) RMs. According to the degree of uncer-
tainty, RMs can be classified as: (i) primary RMs,
(ii) secondary RMs, and (iii) in-house or working RMs.
In order to use RMs for QC, the materials should be
characterised with respect to homogeneity, stability and
the certified property value. All of these parameters are
important characteristics to establish an efficient IQC. In
the case of EQC (PT), homogeneity is essential and
sample stability within the time-scale of the exercise
must be assessed and controlled. When the QC is focused
on qualitative analysis, the use of RMs, sample blank and
matrix standard are crucial to detect false positives and
false negatives, respectively. Although a large number of
RMs are commercially available, most of them are for
quantitative purposes when it is difficult to find sample-
blank RMs. This has meant that in-house RMs have been
used widely in qualitative analysis. In these cases, the
key issue that needs to be considered is the selection of
the material in order to assure homogeneity, stability
and an appropriate or representative matrix. Although
usually the measurement of QC can be carried out by
using in-house materials of adequate homogeneity and
stability, the possibility of using commercially available
sample-blank RMs to validate screening methods is an
interesting point.
4. IQC
The international standard ISO/IEC 17025 points out
that any laboratory shall have QC procedures for moni-
toring the validity of tests. The international standard
also indicates that the resulting data should be recorded
in such a way that trends are detectable, and, where
practicable, statistical techniques shall be applied to
reviewing the results.
Control charts are one of the oldest methods used to
monitor and control routine analyses because of their
simplicity and their usefulness in detecting negative
Trends
trends in analytical work. The objective is to detect
negative effects on results and to identify their potential
source. Although control charts may take a variety of
forms, basically they are a graphic representation on
which the values of the quality characteristic under
investigation are plotted sequentially. Several inter-
national standards of the ISO have addressed control
charts [10–12]. Nevertheless, none of them has addressed
qualitative tests. EUR 20605 EN (2002) describes the
adaptations needed to expand the use of these charts to
qualitative analysis [7].
The principle of control charts is simple and can be
structured in three steps:
(1) take sample during the process;
(2) measure the quality indicator; and,
(3) mark the result on a chart with limits (warning
and action).
Normally, if a CRM is used, the certified value is used
as the assigned value. In using in-house RMs, the
assigned value is the mean of repeated measurements.
Conventionally, warning and action limits are estab-
lished as ±2s and ±3s range, respectively, which corres-
pond to probabilities of 95.5% and 99.7%, respectively,
of the data being normally distributed. Obviously,
warning and actions limits calculated in the above way
should not be used without considering the fit-for-
purpose requirement of the analysis.
These steps are easy to apply to quantitative analysis
but, in qualitative analysis, some problems can appear.
For example, if the binary yes/no response is obtained by
transforming the analytical signal, the limits can be
established, but, if the binary response is provided
directly by the screening method, it is not possible to
establish such limits.
4.1. Simple practical approach
The control chart is the most recommended approach to
use when a binary response is provided directly by the
screening method. For this approach, two samples are
selected (A and B). The samples must be homogenous
and stable and the matrix must be representative of the
matrix of the real samples being analysed. Sample A and
B must have a concentration of analyte slightly lower
and slightly higher than the cut-off or threshold con-
centrations, respectively. Therefore, sample A must
produce a 100% negative response and be used to detect
false positives. Sample B must produce a 100% positive
response and be used to detect false negatives.
Fig. 3 shows a hypothetical example of this simple
practical approach. As can be seen, situations out-of-
control can be easily detected. When the binary re-
sponse is obtained by data transformation, it is possible
to determine the probability–concentration graph. In
this case, it is recommended to establish the concen-
trations of samples A and B from a graph, as shown in
Fig. 4.
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Trends
Check of false positives (A: low concentrations ofanalyte)
Check of false negatives (B:h igh concentrations of analyte)
OUT-OF-CONTROL SITUATIONS
Figure 3. Hypothetical example of a simple control chart used for a qualitative analysis method showing two out-of-control situations.
Figure 4. Selections of concentrations of samples to establish IQC
from a typical probability–concentration graph for a screening
method. Sample A will be used to detect false positives and sample
B to detect false negatives.
4.2. Control charts for blanks
The control charts for blanks are a variation of a
Shewart graph. In this chart, the instrumental signal is
represented as a function of number of the blank control
sample. The objective of these control charts is the same
for qualitative or quantitative methods (i.e., basically to
control the performance of the equipment and the
system), so, from this chart, information about the
contamination of reagents or the state of the instrument
can be obtained. Blank samples can also be used to detect
false positives, although, for that, it is recommended that
a sample with a low concentration of analyte should be
used (e.g., sample A in Fig. 4). In the case of screening
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Trends in Analytical Chemistry, Vol. 24, No. 6, 2005
FALSE ANSWER
CORRECT ANSWER
days
CORRECT ANSWER
FALSE ANSWER
days
method that provides a purely qualitative response, the
use of control charts is limited to detecting false positives.
4.3. Control charts for binary response
The first approach to control charts for binary response
is to use a control chart similar to the common Shewart
control chart. For that, the first step is to select from the
probability–concentration graph two samples at two
concentration levels, as described in Section 4.2 above
(Fig. 4). Afterwards, the reference lines to be used for
control must be defined. A proposal for these reference
lines [7] established the acceptance error level at 5% of
false response, the warning limit at 10% of false response
and the control limit at 15% of false response. Finally,
the false negative and positive rates are determined and
results are graphed in the control chart. It is important
to note that, in this case, the rate of false responses is
represented and not the result of a simple analysis as in
the case of the simple practical approach described in
Section 4.1 above.
Fig. 5 shows a typical control chart for eight daysÕ
monitoring of the rates of false positives and false neg-
atives provided by the test method. As can be seen, in
this example, the majority of error are between the
warning and the acceptance limits.
Taking into account that the objective of this type of
control chart is to detect false responses, rather than the
Shewart chart, the use of a graph similar to the Youden
plot used for quantitative analysis has been recom-
mended [7]. As the objective is to detect both false
positives and false negatives, this control also uses two
control samples, the concentrations of which determine
the unreliability region (Fig. 4). The chart graph com-
prises two axes that define four regions (Fig. 6). Every
point of the group is obtained as result of analysis of the
two samples and is represented in the graph, so each
quadrant of the graph represents a different situation, as
indicated below:
Trends in Analytical Chemistry, Vol. 24, No. 6, 2005
Figure 5. Control chart similar to Shewart chart for monitoring the rates of false positives and false negatives.

Region 1 represents a situation under control.

Region 2 represents a situation in which a false
positive has been detected. Sample B gives a
positive response.

Region 3, in which sample A gives a negative re-
sponse, represent a situation in which false neg-
ative is obtained.

Region 4 represents a double false response. In
this region, a false positive plus a false negative
are obtained.
Fig. 7 shows the control chart used during the anal-
ysis of nitrate in synthetic and real water samples [13].
The authors proposed the use of a blank control sample
to check the negative response and to detect possible
false positives for both screening and confirmative
methods. A sample containing 0.10 lg/L of nitrate was
used to control positives responses in the screening
method. The positive response of the confirmatory
method was checked using a sample containing
0.16 lg/L of nitrate. The control chart obtained pointed
out when any situation out of control was detected.
4.4. Comparison with a confirmatory method
In many cases, a confirmatory method is also used
together with the screening method, following the
so-called vanguard–rearguard principle [14]. The general
Figure 6. Control plot equivalent to Youden plot for IQC of binary
response.
Trends
principle of such a strategy is based on the systematic
analysis of samples using rapid, low-cost vanguard
analytical systems (screening method), which allow the
rapid classification of samples according to a target
characteristic. As a result, only the selected samples
must be processed by a conventional or rearguard
analytical system in order to expand the analytical
information, so the influences of false negatives and false
positives are different. False negatives are most relevant
because the confirmatory or rearguard method is
regularly applied to those samples.
When the analytical system also involves a confir-
matory method, it can be used to control the screening
method. For that, negative samples must be also ana-
lysed by the confirmatory method in a random fashion.
The results obtained for both methods can be repre-
sented in a Youden plot, as in Fig. 8. This IQC can be
introduced by analysing the same samples at different
times or by analysing different samples. In the former
case, the method can also be used to control to perfor-
mance of the confirmatory method. In the latter case,
only differences between both methods can be detected.
Figure 7. Example of an IQC for a screening/confirmative method
for the determination of nitrate in water. Adapted from [13].
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Trends
Figure 8. Control chart for monitoring difference between the
qualitative method of binary responses (vanguard method) and
the confirmatory method (rearguard method).
4.5. Frequency of QC samples
It is the responsibility of the analyst to select an
appropriate level of QC. This level must be selected
taking into account the reliability of the method and the
criticality of the work. As was indicated above, the
samples must by representative of the real problem and
appropriate in terms of homogeneity, stability and reli-
ability. According to QA principles, laboratories should
plan the number of control samples to be used, as well
as the frequency of analysis. In quantitative analysis,
there is wide acceptance of a QC level of 5% (i.e. 1 QC
sample for every 20 samples analysed). For complex
procedures, a level of 20% or even 50% is not unusual.
By contrast, for robust routine methods, a QC level
lower than 5% may be reasonable. In qualitative
analysis, the QC level must take into account the sim-
plicity and the rapidity of the method, which makes it
easy to increase the number of samples analysed.
Table 1 shows recommendations for frequency. In all
cases, it seems reasonable to choose at least two control
samples (one positive and the other negative) to
implement appropriate QC for qualitative methods pro-
viding binary response. Obviously, in order to select the
samples correctly, it is important to know the proba-
bility-concentration graph of the qualitative method.
5. EQC
As was indicated in Section 1, due to the importance of
producing fit-for-purpose data, it is important to dem-
onstrate the accuracy and also the comparability of the
Table 1. Selection and frequency of use of control samples in qualitative analysis
Sample Control of true positive and negative samples
n < 20 Samples with similar matrix Three for series
n > 20 Samples with similar matrix Insert controls every 6 analyses
Samples with different matrix Group samples and insert controls for each matrix
or compare the results with another method
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Trends in Analytical Chemistry, Vol. 24, No. 6, 2005
results. The comparability of results can be demonstrated
by interlaboratory comparisons or PT.
In general, there are different reasons for interlabo-
ratory tests, namely method validation, characterisation
of RMs, and PT. The third and most important for QA
(i.e., PT) aims to obtain an indication of the performance
of the laboratory.
There are several guidelines on the requirements for
PT schemes. Probably, the most important are the guides
from ISO [15,16] and Eurochem [17]. However, these
guides focus on quantitative analysis and they are not
applicable to qualitative analysis.
In 1997, the Analytical Methods Committee of the
Analytical Division of the Royal Society of Chemistry
published an article on handling false negatives, false
positives and reporting limits in analytical proficiency
tests [18]. The authors of the article pointed out that,
when proficiency tests are made with samples contain-
ing little or no measurand, problems occur when
applying the z-scoring system when false negatives and
false positives occur. The article recommends avoiding
the use of terms such as ‘‘not detected’’ and ‘‘less than’’.
It is therefore evident that a new approach for PT
schemes dealing with binary response is needed. As a
starting point, that part of EUR 20605 EN (2002) [7]
that deals with PT schemes of qualitative analysis must
be considered.
5.1. PT schemes
The plan of the PT proposed in the EUR 20605 EN
(2002) [7] is based on the number or percentage of true
responses obtained in the overall assessment of the lab-
oratories. It recommends distributing several samples,
including a set of positive and negative samples. The
simulated example presented in EUR 20605 EN (2002) is
shown in Table 2. As can be seen, 10 samples (three
positives and seven negatives) are distributed to four
laboratories. It is important to ensure that all the sam-
ples have a concentration of analyte outside the range of
concentration established by the unreliability region of
the binary response. Table 2 also simulates the results
for the four laboratories. When a true response is
obtained, a ‘‘0’’ mark is written in accordance with the
z-score system in which ‘‘0’’ is the best score. The mark
‘‘1’’ is used to denote a false response. The report
recommends considering the individual values of the
response obtained for each sample and laboratory as a
partial z-score. The combined values of z shown in Table
Control of blank samples
One for series
One every 15 samples
One every 15 samples
Trends in Analytical Chemistry, Vol. 24, No. 6, 2005 Trends

Table 2. Simulated results of a proficiency testing exercise using a

6. Conclusion
binary response method
QC is an important point to be considered under the
Sample Laboratory Laboratory Laboratory Laboratory
umbrella of enforcing legislation and the new interna-
1 2 3 4
tional standard ISO/IEC 17025. The basis of QC and the
A (+) 0 0 0 1 activities to be carried out in IQC and EQC are well
B (+) 0 0 1 0
established for quantitative analysis, but the basis for
C () 0 1 0 0
D () 0 0 0 1 qualitative analysis needs to be clearly established. This
E () 0 0 0 0 article shows that, with appropriate modifications, the
F () 0 0 0 0 most important points of QC in quantitative analysis can
G (+) 0 0 0 0 be applied to qualitative analysis.
H () 0 0 1 0
I () 0 0 0 1
J () 0 0 0 0
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True responses are marked as ‘‘0’’ and false responses as ‘‘1’’.
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