In the Laboratory
Determination of Anionic Surfactants Using Atomic
Absorption Spectrometry and Anodic Stripping
Voltammetry
Richard John and Daniel Lord
School of Applied Science, Griffith University, Gold Coast Campus, Parklands Drive Southport
Gold Coast, QLD, 4214, Australia; *R.John@mailbox.gu.edu.au
Atomic absorption spectroscopy (AAS) and anodic strip-
ping voltammetry (ASV) are techniques typically restricted
to metal analysis. An experiment developed for our under-
graduate teaching program demonstrates the utility of these
techniques for the determination of organic species, in this case,
the indirect analysis of anionic surfactants. The method involves
formation of an extractable complex between the synthetic sur-
factant anion and the bis(ethylenediamine)diaquacopper(II)
cation. This complex is extracted into chloroform and then
back-extracted into dilute acid. The resulting Cu(II) ions are
determined by AAS and ASV. Students are required to deter-
mine the concentration of a pre-prepared “unknown” anionic
surfactant solution and to collect and analyze a real sample
of their choice.
The experiment was designed as a final-year under-
graduate laboratory for the subject Analytical Chemistry
within our Environmental Science degree. It demontrates
the following analytical principles:
indirect analysis
compleximetric analysis
liquid–liquid (solvent) extraction
back extraction (into dilute acid)
analytical recovery
analysis using flame-AAS and ASV, two of the most
popular methods for trace metal analysis
In their write-up, students are expected to discuss each
of the above principles and make a critical comparison of
metal analysis by AAS and ASV. The experiment expands on
a previous publication in this Journal for the determination
of anionic surfactants, which employed a colorimetric end-
point for the analysis (1).
Background
Anionic surfactants are widely used for industrial and
domestic purposes, including laundry powders, dish-washing
liquids, shampoos, emulsifiers for industrial solvent cleaners,
and agricultural and horticultural chemical delivery systems.
Although most anionic surfactants in use today are biode-
gradable and essentially nontoxic to humans (2), a need still
exists for effective monitoring of these compounds. Aside from
the visual pollution (in the form of foaming) associated with
surfactant use, the environmental impact of surfactant release
in natural water systems can be much more serious. For ex-
ample, several authors have reported impaired respiratory
function in fish caused by damage to the gill epithelium at
concentrations below 3 mg L{1 (3–6 ). In addition, damage to
chemoreceptors of fish at concentrations as low as 0.5 mg L{1
1256 Journal of Chemical Education • Vol. 76 No. 9 September 1999 • JChemEd.chem.wisc.edu
W
has been shown to impair feeding behavior (7 ). While the
more active marine adults (such as fish) are more susceptible
to the presence of anionic surfactants than less active adults
(such as crustaceans), the developmental stages of all forms
of marine life are extremely susceptible to even trace levels
of these compounds. In particular, invertebrate and fish
larvae show susceptibility to anionic surfactant concentra-
tions of 0.05 mg L{1 (8) and 0.16 mg L{1 (9), respectively.
Analysis of Anionic Surfactants
The APHA standard method for the determination of
anionic surfactants is the Methylene Blue Active Substances
(MBAS) method (10). This method employs the methylene
blue cation, which forms a 1:1 ion pair with the surfactant
anion. The ion pair is extracted into chloroform and deter-
mined spectrophotometrically. Unfortunately, the MBAS
method lacks specificity because many sulfonates and simple
inorganic anions such as cyanate, nitrate, thiocyanate, sulfide,
and chloride also form extractable compounds with methylene
blue, resulting in positive interferences. For this reason, the
MBAS method for anionic surfactants was replaced as the
Australian standard method by a method that uses the
bis(ethylenediamine)diaquacopper(II) cation as the extracting
agent (11). This ion permits selective quantitative extraction of
surfactant anions into an organic solvent (1). The surfactant
anion reacts with the aquated bis(ethylenediamine)copper(II)
ion (12) and is extracted into chloroform as a neutral inner
sphere complex (eq 1). The [Cu(en)2(RSO2O)2](org) com-
plex is back-extracted with dilute acid, releasing free Cu(II)
into the aqueous phase. The copper concentration is then
determined using AAS and ASV.
[Cu(en)2(H2O)2]2+(aq) + 2RSO2O{(aq) →
(1)
[Cu(en)2(RSO2O)2](org) + 2H2O(aq)
en = ethylenediamine (H2NCH2CH2NH2)
R = hydrocarbon group (C12H25C6H4)
In contrast to the Australian standard method for anionic
surfactant analysis, which utilizes flame AAS, the instrumen-
tation required by ASV is simpler and more robust, without
the need for specialized gases and extraction systems. The
simple instrumentation means that very little operator ex-
pertise is needed, making it ideal for undergraduate students.
Unlike ASV, flame-AAS cannot be used to measure copper
concentrations below 0.02 ppm, and the calibration curve
for ASV generally extends over more orders of magnitude
than AAS. Of its spectroscopic competitors, only flameless
AAS is able to achieve the same sensitivity as ASV, but at a
much higher cost.
 In the Laboratory

0.018 A = 0.0349 x concn - 5 x 10-4) Experimental Procedure

0.016 The preparation of reagents and full experimental pro-
cedures are based on the Australian standard method for
0.014
anionic surfactant analysis, AS 3506 (11). A brief outline of
0.012 the procedure and required equipment is given below. Full
A324.8nm

0.010 details can be found in the online lab documentation.W

0.008 General Procedure
0.006 Students are required to prepare anionic surfactant cali-
0.004
bration standards of 0.05, 0.1, 0.2 and 0.5 mg L{1 by serial
dilution of a pre-prepared 1000 mg L{1 stock solution. The
0.002
anionic surfactant standard used in this experiment is
0 dodecylbenzene sulfonic acid.
0 0.1 0.2 0.3 0.4 0.5 Foam formation should be avoided because the concen-
Concentration (ppm) tration of surfactant in the foam phase is significantly greater
Figure 1a. AAS calibration curve for Cu(II) standards.
than in the aqueous phase. Compressed air can be used to
collapse any foam that forms.
Two hundred milliliters of each standard is pipetted into
250-mL separating funnels. The copper ethylenediamine
0.014 A = 0.0271 x concn - 2 x 10-5) reagent (pre-prepared) and chloroform are then added. After
0.012 liquid–liquid extraction the chloroform layer is run off into
a centrifuge tube and centrifuged for 2 min at 2500 rpm to
0.010 achieve complete phase separation. (Alternatively, phase sepa-
ration can be achieved by allowing the chloroform extracts
A324.8nm

0.008 to stand for 20 min.) Ten milliliters of the clarified extract is

then pipetted into a 25-mL stoppered measuring cylinder and
0.006
10.00 mL of 0.1 M HCl is added. This is shaken and allowed
0.004
to stand for 20 min, after which the aqueous layer is removed
and analyzed for Cu(II) by AAS and ASV. A blank solution
0.002 is also analyzed by using 200 mL of deionized water in place
of the surfactant standards.
0 Students are required to determine the concentration
0 0.1 0.2 0.3 0.4 0.5
of a pre-prepared “unknown” (the concentration is known
Concentration (ppm) to the lab demonstrators) dodecylbenzene sulfonate solu-
Figure 1b. AAS calibration curve for dodecylbenzene sulfonic acid tion by comparison with standards. In addition they are
standards. required to collect and analyze a real sample of their choice
(see below).
Sampling
Samples should be collected in glass bottles and, as soon
as practicable, filtered through a 0.45 ± 0.05-µm cellulose
35
Ip = 68.031 x concn + 0.1964 acetate membrane filter. The filtered sample should be stored
30 at 4 °C and analyzed within 48 hours of collection. With
concentrations below 1 ppm, adsorption onto the wall of the
25 sample container will lead to reduced surfactant levels in the
bulk aqueous phase. Addition of alkali phosphate will mini-
20 mize adsorption errors (12).
Ip / µA

Three readily available sources of anionic surfactant

15
samples on our campus were used in this experiment: the
10
water from (i) the sink of a cafeteria kitchen, (ii) the dish-
washer from the same kitchen, and (iii) a domestic washing
5 machine at the end of its wash cycle. These samples, while
not typical environmental samples, were convenient examples
0 to demonstrate the principles of sampling and analysis required
0 0.1 0.2 0.3 0.4 0.5 in this experiment. It should be noted, however, that the
Concentration (ppm) anionic surfactant concentrations in these samples exceeded
Figure 2. ASV calibration curve for dodecylbenzene sulfonic acid the calibration standards, necessitating suitable dilution of
standards. the samples by the students.

JChemEd.chem.wisc.edu • Vol. 76 No. 9 September 1999 • Journal of Chemical Education 1257

 In the Laboratory

Results and Discussion Table 1. Concentration of

Laborator y-Prepared "Unknown"
The ratio of copper to surfactant in the extracted complex Samples
is 1:2. Given the experimental procedure used and assuming Result/ppm
True value/
100% extraction efficiency, a sample containing 0.1 ppm of Student No.
ppm AAS ASV
surfactant should yield a Cu(II) concentration of 0.13 ppm
after back-extraction into acid. Figure 1a shows a typical cali- 1061923 0.10 0.12 0.13
bration curve obtained by students for the determination of 593106 0.20 0.20 0.21
Cu(II) standards by AAS. The students use this curve for com- 455969 0.50 0.49 0.48
parison with the calibration curve obtained for analysis of
the dodecylbenzene sulfonic acid standards (Fig. 1b).
The ratio of the slopes for these two curves (Cu/surfactant) Table 2. Concentration of "Real"
is (0.0349/0.0270), or 1.29. This equates with the ratio expected Samples Collected by Students
for 100% extraction efficiency (0.13 ppm/0.10 ppm = 1.30).
Result/ppm
The students use this procedure as a check to ensure they Sample
have 100% (or close to it) recovery of surfactant. AAS ASV
Figure 2 shows a typical calibration curve obtained for Commercial sink 121 122
dodecylbenzene sulfonic acid standards using an ASV finish. Commercial dishwasher 5.16 5.49
A similar check on percent recovery could be performed using Domestic washing machine 70.0 71.7
ASV analysis of Cu(II) standards, although we do not make
this part of our procedure.
The results of three typical student analyses of laboratory
prepared unknown surfactant samples are shown in Table 1.
As the table shows, we find that the AAS and ASV results
correlate very well and errors tend to result from students’ Literature Cited
technique (dilutions, extractions, preparation of standards,
etc.) rather than from the end analyses. 1. Crisp, P. T.; Eckert, J. M.; Gibson, N. A. J. Chem. Educ. 1983,
Students are also required to collect a sample containing 60, 236–238.
anionic surfactant from one of three sites on campus. Table 2. Swisher, R. D. Surfactant Biodegradation, 2nd ed.; Dekker: New
2 shows the results obtained from three student analyses. York, 1987.
Again, the AAS and ASV results show good correlation. 3. Swedmark, M.; Braaten, B.; Emanuelsson, E.; Granmo, A. Ma-
Details of interferences for anionic surfactant analysis of rine Biol. 1971, 9, 183.
real samples can be found in ref 12. These are also outlined 4. Schmid, J.; Mann, H. Nature 1961, 192, 625.
in the lab documentation. Specific details concerning the 5. Swisher, R. D.; O’Rourke, J. T.; Tomlinson, H. D. J. Am. Oil
ASV and AAS parameters are also provided in the lab Chem. Soc. 1964, 41, 746.
documentation.W 6. Eisler, R. Am. Fisheries Soc. Trans. 1965, 94, 26.
7. Bardach, J. E.; Fujiya, M.; Hall, A. Science 1965, 148, 1605.
Acknowledgments 8. Renzoni, A. Arch. Oceanogr. Limnol. 1975, 18(2), 99.
9. Lesyuk, I. I.; Kostyuk, A. O.; Lemishko, A. A.; Reshetito, S. G.;
We wish to thank Mark Imisides and Melinda John for Kotsqumbuas, I. Y. Vopr. Ikhtiol (Russia) 1983, 23(b), 993.
helpful discussions in the preparation of this experiment. 10. Standard Methods for the Examination of Water and Waste Water,
Many thanks also to Greg Hope, Paul Duckworth, and AD 16th ed.; American Public Health Association: Washington, DC,
Instruments for advice and the use of the Maclab/4e inter- 1985.
face and potentiostat. 11. AS 3506. Waters. Determination of Filtrable Anionic Surfactants—
Copper–Ethylenediamine Flame Atomic Absorption Spectrometric
Note Method; Standards Association of Australia: N.S.W Australia,
W
Supplementary materials for this article are available on JCE 1987.
Online at http://jchemed.chem.wisc.edu/Journal/issues/1999/Sep/ 12. Crisp, P. T.; Eckert, J. M.; Gibson, N. A. Anal. Chim. Acta 1975,
abs1256.html. 78, 391–396.

1258 Journal of Chemical Education • Vol. 76 No. 9 September 1999 • JChemEd.chem.wisc.edu