Environmental Sampling
for Trace Analysis
Ray E. Clement
Ontario Ministry of the Environment
Laboratory Services Branch
125 Resources Rd.
P.O.Box 213
Rexdale, Ontario, Canada M9W 5L1
Hundreds of millions of dollars are
spent worldwide each year on envi -
ronmental issues-based directly on
the results of chemical analyses. One
would think that students in chemis-
try and engineering programs, who
will be employed in the environmen-
tal field, should receive basic train-
ing in evaluating the reliability of
such chemical analyses. However, it
is unfortunately true that far too few
of these graduates will have such
training. Many new environmental
programs being introduced in our
colleges and universities emphasize
the study of environmental issues
r a t h e r t h a n t h e scientific tools
needed to investigate these issues.
Few undergraduate programs em -
phasize the type of laboratory in-
struction in analysis that is needed
to understand how reliable chemical
data are generated. Although most
analytical textbooks and courses dis -
cuss the importance of accurate sam-
pling, students are seldom offered
practical examples of the conse-
quences of poor sampling. They may
think that an analytical result gener-
ated by a million-dollar state-of-the-
art instrument is a number “carved
in stone.” And because such analyses
are expensive, they believe there is
no need for replication.
Can students learn the basic prin-
ciples of environmental sampling and
analysis from a simple classroom ex-
periment? To find out, a sampling ex-
periment was conducted at Mohawk
College (Canada) in a class of stu-
dents who were not chemistry ma-
jors. Although the experiment was
originally designed to illustrate sam-
pling difficulties and analytical er-
1076 A ANALYTICAL CHEMISTRY, VOL. 64, NO. 22, NOVEMBER 15,1992
A oms and molecules. Green was used
Classroom
Experiment
You Can
Sink Your
Teeth Into!
rors that can occur in trace environ-
mental determinations, it became
apparent that it could also be used to
describe many analytical concepts-
including the importance of sample
treatment and the need for selective
detection. Most important, it clearly
demonstrates the reality of errors in
analytical measurements. Because
the experiment is simple, interactive,
and directly analogous to the analyt-
ical principles described above, it is
highly effective and even enjoyable.
Design of the experiment
Nest16 manufactures a candy product
called Smarties (available in Cana-
da), which are button - shaped choco-
late candies that have multicolored
h a r d outer coatings. Individual
Smarties are relatively uniform in
shape and weight, and in appearance
differ only in the color of the outer
coating. (Any brand of multicolored
candy can be substituted.) In the
sampling experiment, different colors
were used to represent different at-
to signifjr PCB molecules, blue repre-
sented Pb, brown designated Fe, and
red symbolized chlorinated dioxin
molecules. In addition, purple and
pink represented unknown mole-
cules. A soil matrix was depicted by
combining yellow and orange can-
dies. The manufacture of a sample
where the matrix predominated was
difficult, because the Smarties, as
marketed, are fairly evenly distrib-
uted among the various colors. To
avoid having to purchase kilogram
quantities of Smarties, a second type
of candy similar in appearance to
Smarties was purchased. This candy,
called Reese’s Pieces (manufactured
by Hershey), comes in orange, yellow,
and brown. The brown Reese’s Pieces
also represented Fe.
After combining the Smarties and
Reese’s Pieces, the number of candies
of each color were counted. Because
the individual molecules and compo-
nents of the soil matrix are almost
uniform in size and shape, we as-
sumed for the purpose of this experi-
ment that the molecular weights of
all the individual candy pieces were
the same. Therefore, by counting the
number of candies of the same color,
the concentration of the molecule or
atom represented by that colored
candy could be determined in units of
parts per hundred (pph) (Table I).
Two volunteers from the class were
selected to run the experiment. Stu-
dent A was instructed to sample the
contents of the plastic container in
which the candies were brought to
class by grabbing a small handful.
After t h e first grab sample was
taken, student A was told to make a
second small grab and to keep this
sample separate from the first. Stu-
dent B was given the same instruc-
tions, except he was told to take
large handfuls. The class was then
told to determine the concentrations
of all of our target analytes in parts
per hundred-based on the four grab
0003-2700/92/0364-1076A/$03.00/0
0 1992 American Chemical Society
REPORT
samples. An overhead projector was
used to record the results.
Effect of sample size and
replication on precision and
accuracy
The actual experimental data from
this classroom experiment are shown
in Table 11. Quite a large difference
can be seen in the replicate results
from student A, who collected total
sample sizes of seven candies for
each replicate. Unfortunately, the
analyte with the highest concentra-
tion, Fe, was not detected in either
replicate sample. PCBs, Pb, and the
two unknown substances were de-
tected in the first sample, each at 14
pph; only one of the unknowns was
found in the second sample, at 28
PPh.
Clearly, the differences between
the replicates are significant. The re-
sult of selecting a small sample size
was to increase (make worse) the de-
tection limit of the determination.
The comparison of individual results
to the actual values seems pretty
poor. However, when the averages of
the values of the two determinations
are taken, it can be seen that the es-
timates are much closer to the actual
values; generally the estimated con-
centrations are within a factor of
about 2-not too bad for trace analy-
sis. Only the results for Fe and the
unidentified pink analyte are way
off. In the case of dioxin, the method
is not sensitive enough to detect such
an ultratrace analyte.
Real - life trace environmental
analysis is conceptually not so differ-
ent from this example; when method-
ology is used that can barely detect
analytes, it is not uncommon for the
analytes to be observed in some sam-
ples but not in others, even though
all samples were thought to be the
same. For example, in a six-labora-
tory round-robin study of chlorinated
dibenzo -$-dioxins and chlorinated
dibenzofurans introduced into blank
water samples, a significant varia-
tion in laboratory results was noted
(1). The reported concentrations of
:ntroduced at a con-
concentrations by using a
1078 A ANALYTICAL CHEMISTRY, VOL. 64, NO. 22, NOVEMBER 15,1992
greater sample size. In these larger
handfuls each analyte in the
was detected in both replicates ex-
cept for “dioxin,’’ which was observed
in only one sample. The concentra-
tions of all analytes are more accu-
rate and more precise for the large-
sample - size experiment with the
single exception of PCBs, where the
mean concentration as determined
from the first sampling experiment
(small sample size) was closer to the
actual value. As in the first sampling
experiment, the mean estimates from
two determinations were closer to
the correct values than the estimates
from an individual sampling event
(with the single exception of PCBs,
where both replicates from student B
produced low estimates). The close-
ness of the mean analyte values from
student B to the actual concentra-
tions is quite reassuring.
Sample preparation and analyte
detection
A number of important concepts re-
lating to sample preparation and an-
alyte detection can be illustrated by
this simulated soil- sampling experi-
ment. Before sampling, the class was
asked if any special sample prepara-
tion were needed. Almost everyone
suggested t h a t the sampling con-
tainer should be well mixed before
samples were withdrawn. It was also
important (for this specific experi-
ment) that the sampler take a “blind”
sample; otherwise, the sample se
lected could be b’
sampler’s favorite
is not recommende
contaminant plume that is emitted
from a plant.
Some s a m p l e “cleanup” w a s
needed before analytes could be accu-
rately detected. In this experiment,
cleanup consisted of sorting the can-
dies into groups according to color,
after which they could be assayed by
counting. For the large sample, an
accurate quantitative determination
could not be made without this sort-
ing step, although without cleanup it
is still possible to do a qualitative de-
termination of the analytes present
by recording the different colors ob-
served.
The physical separation of the can-
dies into different colors is analogous
to various chromatographic separa-
tion methods that are the basis of
most real cleanup schemes. Note that
the cleanup is more difficult to per-
form with the large sample. In real
life, it is not difficult to overload
cleanup methods by choosing sample
sizes greater than those for which
the cleanup is designed. I n such
cases, choosing a large sample size
will increase rather than reduce the
detection limits ac
experiment the dete
not good for the sm
taken initially, but
tection took more time to complete.
The detector in this experiment
was the human eye. By observing the
various colors in the sample, the dif-
ferent analytes were identified (qual-
itative analysis). By counting the
number of candies in each color
group, quantitative estimates of
 Table II. “Contaminant” dat;
~~
their concentrations were deter -
mined. A serendipitous illustration of
some of the difficulties in analyte de-
tection could be made because the
test sample was made up of a mix-
ture of Smarties and Reese’s Pieces.
Although they appeared to be simi-
lar, Reese’s Pieces are a bit smaller
than Smarties. This size difference is
not critical for the “soil matrix” itself,
but it is important for the detection
of the brown-colored candies used to
represent Fe. Therefore the method
of detection (color) by itself cannot
distinguish between the two brown
analytes. A determination of physical
properties (size) is also needed to
avoid i overestimation of Fe in the
ten important to know
form of an analyte is present in the
environment and, as illustrated in
this experiment, this can be very dif-
ficult. In this case, a second method
of detection (taste) can be used to
distinguish between the Smarties
and Reese’s Pieces. (Note that taste
is destructive detection; observation
of color is nondestructive detection.)
Other observations and
limitations
At this time some readers may be
noting that the concentrations de-
ND
28
tected are inaccurate because the
students conducted sampling without
replacement. In other words, the can-
dies taken from the container by the
first sampler were not replaced; con-
sequently, the actual concentrations
of the candies in subsequent sam-
plings were affected by this omission.
This is true, but by selecting a large
original population size, this factor is
not significant (at least for the pur-
poses of this experiment).
In real environmental sampling,
the amount of analyte removed from
the area sampled (e.g., field, lake, or
ambient air) is trivial wheh com-
pared with the total amount of ana-
lyte present. In fact, in real environ-
mental sampling one samples such a
small amount of the whole area to be
tested that a single sample quite
likely is not representative of the
area under investigation. Therefore
The sampling experiment with
candies represents an idealized situ-
ation in which the entire population
ANALYTICAL CHEMISTRY, VOL. 64, NO. 22, NOVEMBER 15, 1992
.4
4.2
is present in a plastic container, and
therefore the concentrations of the
various analytes could be determined
exactly. In the environment, it is not
possible to know the exact concentra-
tion of any analyte in the population.
The best we can do is to estimate an-
alyte concentrations by using a rig-
orous and careful sampling program,
with judicious use of replication to
determine the precision of our esti-
mates. The population used in this
experiment is also static; the exact
numbers of the various color
dies could change only i
removed by accident or
environment represents
situation in which anal
trations depend on wea
tions, time of day, type
and natural processes occ
mine concen-
must find the
detector response to analytes by
evaluating standards prepared with
known concentrations of these ana-
lytes. For this soil-sampling experi-
9 1079 A
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REPORT
~ ~ ~~

ment, the concentrations of the two
unknown analytes could not be de-
termined until their identities were
established.
An interesting problem was pre-
sented to the students after the ini-
tial sampling experiment was com -
pleted. They were asked to sample
the population to find a single poi-
soned candy that had been thrown
in. In the absence of a specific detec-
tor that could selectively detect the
poison (nobody volunteered to per-
form a taste test!), the students fa-
vored disposal of the entire popula-
tion. This specific situation seldom
occurs in an environmental analysis,
but it is now obvious that the objec-
tives of a sampling experiment
should be clearly stated before any
work is performed. The methods se-
lected for any application depend on
the specific results desired.
Conclusions
In the 30 minutes it took to conduct
this experiment, students received a
better understanding of the princi-
ples of environmental sampling and
analysis than if they had read a text-
book on the subject. Hands-on exper-
iments capture t h e interest and Suggested reading
attention of the students and encour- Principles of Environmental Sampling; Keith,
age them to use their imaginations. L. H., Ed.; ACS Professional Reference
Nobody is suggesting that the stu- Books; American Chemical Society:
dents are now experts in environ- Washington, DC, 1988.
mental sampling and analysis, but Keith, L. H. Environmental Sampling and
Analysis: A Practical Guide; Lewis Pub-
after this experiment they are better lishers: Chelsea, MI, 1992.
prepared t o consider the details of
such methods used for environmen-
tal investigations. The most impor -
tant lesson is t h a t sampling and
analysis methods are used to esti-
mate the concentrations of analytes,
and the answers obtained are subject
to error. Proper methods are re-
quired to give the most accurate and
precise results.
After the experiment and discus-
sion, the remaining candies were
passed around the class for individ-
ual sampling and detection by taste.
The students unanimously agreed Ray E. Clement received a Ph.D. in ana-
that the soil - sampling experiment lytical chemistry (1981)from the Univer-
was literally one they could sink sity of Waterloo under the supervision of
their teeth into. F. W. Karasek. He then joined the On-
tario Ministry of the Environment where
Reference he is a senior scientist in the R&D depart-
ment. He has authored more than 100
(1)Tashiro, C.; Clement, R. E.; Davies, S.; publications, most of which concern trace
Oliver, B.; Munshaw, T.; Fenwick, J.;
Chittim, B.; Foster, M. G. Chemosphere determination of chlorinated dioxins and
1990,20(10-12), 1313-17. jkrans in environmental samples.

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