HUI-FENG ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 84, NO.

6, 2001 1781
ENVIRONMENTAL ANALYSIS

Application of Membrane Filters for Spectrophotometric

Determination of Cationic Surfactants in River Water and
Sediment
SUN HUI-FENG, MIWA TAKAMORI, NORIKO HATA, ISSEI KASAHARA, and SHIGERU TAGUCHI
Toyama University, Faculty of Science, Gofuku 3190, Toyama 930-8555, Japan

Cationic surfactant (CS+) in urban river water and

sediment was extracted and determined spectrophotometrically with 2 membrane filters. The CS+
in the water samples, mostly in the form of an ion
associate with the coexisting anionic surfactant
(AS), was collected on a polytetrafluoroethylene
(PTFE) membrane filter and eluted with methanol.
Bromphenol blue (BPB), hydrochloric acid, and
water were added to the methanol solution successively, and the mixed solution was filtered through
a mixed cellulose ester membrane filter. The
CS+-BPB ion associate, formed by a counter ion
exchange, was collected on the filter and dissolved
into N,N-dimethylformamide (DMF) together with
the mixed cellulose ester membrane filter. After
addition of 2 drops of triethanolamine, the
absorbance of the DMF solution was measured.
The CS+ in sediment samples was extracted with
methanol by ultrasonic irradiation; the methanol
solution was then passed through a PTFE membrane filter and evaporated to dryness. The CS+
was redissolved in a small amount of methanol.
For water samples, recoveries and relative standard deviations for 0.30 mM benzyldimethyltetradecylammonium ion, a standard material, were
93 and 5%, with a detection limit of 0.02 mM.
Concentrations of CS+ in sediments were much
higher than those in water samples, indicating that
CS+ is adsorbed on the surface of the sediment.

ationic surfactant (CS+) is commonly used in fabric

softeners, disinfectants, emulsions, and human hair
cosmetics (1). The use of CS+ has been increasing recently; the production of CS+ in Japan was around 70 000 tons
in 1997 (2). Although CS+ is not as widely used as the anionic
surfactant [AS; 540 000 tons produced in 1997 (2)], its toxicity
is about 10 times higher than that of AS (3). In areas without
waste water treatment facilities, CS+ is directly discharged into
the environmental water and can disturb the ecological balance

Received November 2, 2000. Accepted by JS February 22, 2001.

Corresponding authors e-mail: taguchi@sci.toyama-u.ac.jp.

due to its toxicity to aquatic organisms (4, 5). Therefore, a rapid

and reliable analytical method is needed for the determination of
CS+ in environmental samples.
The most widely used technique for commodity and water
samples is solvent extraction followed by spectrophotometry,
which is based on the formation of ion associate with anionic
dyes (D). Scott (6) used orange II to form an ion associate with
CS+ and extracted it into chloroform. Disulfine blue VN-150 and
bromphenol blue (BPB) were proposed as anionic counter ions
by Longman (7) and Sakai and Ohno (8). We proposed a simple
and rapid method using bis[2-(5-bromo-2-pyridylazo)-5-(Npropyl-N-sulfopropylamino) phenolato]cobaltate(III) as a counter ion (9). However, some organic solvents are very toxic and
must be used under strict control. Furthermore, these methods
suffered from the AS, which easily formed a CS+-AS ion associate because of their high ion association constant (10), thus preventing the formation of CS+-D, and could not be directly applied to samples in the aquatic environment, where AS exists at
much higher concentrations than CS+ (9, 11). A 2-step solvent
extraction method proposed by Nakamura et al. (12) could avoid
the interference from AS. The solvent used in that method,
1,2-dichloroethane, is also very toxic. Liquid chromatography
(LC), capillary electrophoresis (CE), flow injection analysis
(FIA), and solid-phase extraction methods have recently been
reported (1316); however, the LC and CE methods are not
applicable for analysis of CS+ in environmental samples because of AS interference. The FIA method is simple and
rapid, but its sensitivity is still very poor.
In our previous study (16), 2 membrane filters were used
for preconcentration and spectrophotometric determination
of CS+ in environmental water. Only a small volume of
N,N-dimethylformamide (DMF) was used in that method.
The toxicity of DMF is much lower than other organic solvents which are commonly used in the solvent extraction
methods, e.g., the LD50 for DMF (used orally in rats) is
1/100 that of 1,2-dichloroethane (17). The methyl orange
(MO) which acted as a counter ion in that method, however,
was not ideal because of its lower sensitivity for environmental water samples. We proposed a transparent membrane filter for the solid-phase spectrophotometric determination of CS+ in water (18). Although this method is simple
and solvent-free, it was still not sensitive enough.
River sediments basically carry negative surface charge,
and it is reasonable to conclude that CS+ tends to be adsorbed

1782 HUI-FENG ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 84, NO. 6, 2001

Figure 1. Molecular structure of cationic surfactants.

onto the sediment surface. As described above, most CS+ in

environmental water seems to be present as the ion associate
CS+-AS. The CS+-AS ion associates were easily adsorbed
onto the sediments by hydrophobic interaction. Adsorption of
CS+ onto sediment may influence its mobility, toxicity, and
decomposition rate. Utsunomiya et al. (19) found that the decomposition rate of AS in sediment decreased seriously when
CS+ was present. It is important, therefore, to determine the
concentration of CS+ in sediments in order to understand the
behavior of CS+ in aquatic environment. The analytical
method for CS+ in sediments, however, has not been reported.
This study presents a method for the separation and spectrophotometric determination of CS+ in both water and sediment samples by using 2 membrane filters. The method is partially based on one previously reported (16). The BPB,
proposed as a counter ion in this study, improved the analytical sensitivity 4-fold, and effectively avoided the interference
from chlorophyl which was co-extracted with CS+ from sediment samples. Recovery of 0.30 M benzyldimethyltetradecylammonium ion in water samples was 93%, with a detection
limit of 0.02 M. This method was applied to the determination of CS+ in aquatic environments, and the results indicated
that CS+ is quite easily adsorbed onto sediment surfaces.
Experimental

Reagents and Apparatus

(a) Benzyldimethyltetradecylammonium
chloride
dihydrate.(Zephiramine, Zeph+Cl2H2O, analytical
grade) was obtained from Dojindo (Kumamoto, Japan). It
was used as the standard material of a cationic surfactant
because of its high purity, high dissolution to water, and
well-known structure. The standard cationic surfactant
was dissolved in water without further purification.

(b) Other cationic surfactants.Benzethonium chloride,

distearyldimethylammonium chloride, cetyltrimethylammonium
chloride, trimethylstearylammonium chloride, benzyldimethylstearylammonium chloride, and cetylpyridinium were purchased
from Tokyo Kasei (Tokyo, Japan). The molecular structures of
these compounds are shown in Figure 1.
(c) Sodium lauryl sulfate (SDS).Specially prepared reagent was obtained from Nacalai Tesque (Kyoto, Japan).
(d) Stock solution of 1mM SDS.Prepared in 0.1M HCl
and kept at 4C.
(e) Bromphenol
blue
(BPB,
analytical
grade).Purchased from Wako (Osaka, Japan). It was dissolved in 0.03M NaOH followed by addition of 1M HCl to adjust the pH of the solution to 56. The solution was diluted to
5mM with water, and then purified by passing it through an
Advantec Toyo glass fiber filter (Tokyo, Japan). The BPB solution was kept at 4C in the dark.
(f) Methanol and N,N-dimethylformamide.Analytical
grade; also purchased from Wako.
(g) Membrane filters.A polytetrafluoroethylene (PTFE)
membrane filter (0.1 m pore size, 25 mm diameter) and a
mixed cellulose ester membrane filter (0.45 m pore size,
25 mm diameter) were obtained from Advantec Toyo.
(h) Equipment.A
Hitachi
Model
U-2001
spectrophotometer (Tokyo, Japan) equipped with 1 cm glass
cell and a Horiba Model D-12 pH meter (Kyoto, Japan) were
used to measure the absorbance and the pH, respectively. An
Advantec Toyo Model KG-25 filtration apparatus was used under vacuum with an aspirator. A SIBATA Model SU-30 ultrasonic generator (Tokyo, Japan) and a Bchi rotary evaporator
(Flawil, Switzerland) were used for sediment samples handling.

Sample Collection and Preservation

River water samples were taken from the Furu and Jinzu
rivers and from 2 other rivers located in Toyama, Japan. The
acidity of the water samples was adjusted to pH 2 by the addition of HCl. River sediment was taken from the Furu River.
Sediment on the top 5 cm layer was collected, passed through
a 16-mesh sieve, centrifuged, and dried at 110C for 2 h. The
dried sediment samples were passed through a 16-mesh sieve
once again, and kept at room temperature.

Extraction and Preconcentration of CS+ from Water

and Sediment Samples
River water (100 mL) adjusted to pH 2 was placed in a
100 mL Erlenmeyer flask. A mL volume of 1mM SDS was
added to completely convert the CS+ into CS+-AS ion associate.
The water sample was heated on a hot plate at ca 100C for
30 min, and then set aside at room temperature. The suspending
particles in the river water were precipitated during that process.
A glass fiber filter was put on the top of a PTFE membrane filter.
The water sample was then passed through the 2 piled filters. The
precipitate was mostly retained on the glass fiber filter, which
helped to avoid clogging of the PTFE filter. The filters were
washed with 5 mL distilled water and air-dried. The CS+-AS
adsorbed on the precipitate, and the PTFE filter was eluted into a
test tube with 2 mL methanol.

HUI-FENG ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 84, NO. 6, 2001 1783

Figure 2. Structure of BPB ion.

Ultrasonic Extraction and Preconcentration of CS+

from Sediment Sample
A 50 g amount of dried sediment and 200 mL methanol
were placed in an Erlenmeyer flask. The flask was irradiated by
ultrasonic wave for 60 min, and the solution was filtered
through a PTFE membrane filter. The sediment was transferred
into another 200 mL methanol, irradiated, and filtered once
again. The 2 filtrates were combined, transferred to an evaporation flask, and evaporated to dryness on a rotary evaporator.
Methanol (10 mL) was added to the evaporation flask and set
aside for 60 min to completely dissolve the CS+ adsorbed on the
wall of the flask; 2 mL of this methanol solution was then transferred into a test tube for the following procedures.

Spectrophotometric Determination of
Preconcentrated CS+ in Methanol Solution
For both cases, 2 mL BPB solution and 2 mL 4M HCl were
added to the test tube. The mixture was vigorously shaken for
about 10 s and set aside for 10 min. The solution in the test tube
was diluted to 25 mL with water and set aside for another
10 min; the diluted solution was then passed through a mixed
cellulose ester membrane filter. The filter was washed with
5 mL 0.1M HCl. The filtration area was cut off and dissolved
into 5 mL DMF. Two drops (ca 0.08 mL) triethanolamine were
added to the DMF solution, and the absorbance of the DMF solution was measured at 603 nm against a reagent blank.

maximum absorption at 603 nm, whereas chlorophyl has nearly

negligible absorption. Furthermore, the sensitivity of BPB was
much higher than that of MO. For these reasons, BPB was chosen as the counter ion for both water and sediment samples.
In acid solution, the BPB ion takes one negative charge
and, in neutral or basic solution, the BPB ion releases a proton
and becomes a divalent anion (Figure 2). It is well known that
in a hydrophobic interaction, a univalent ion is stronger than a
higher valent ion in water (10, 20). Therefore, HCl and water
were added to the methanol solution to form the stable
CS+-BPB ion associate, which was easily retained on the
mixed cellulose ester membrane filter.
Compared with the acid form, the basic form of BPB gives
a much higher absorptivity and the maximum absorbance
shifts to 603 nm (Figure 3). In this work, triethanolamine was
added to the DMF solution to change the BPB to a basic form
before determination.
The BPB and HCl concentration in the final 25 mL solution
were set at 0.4mM and 0.3M, respectively. Such concentrations ensured quantitative counter ion exchange between AS
and BPB and collection of the formed CS+-BPB ion associate on the membrane filter. The test tubes were vigorously
shaken for about 10 s to ensure complete mixture after addition of BPB and HCl to the methanol elute. The collection rate
of the CS+-BPB ion associate will decrease if the BPB concentration and shaking time are <0.3mM and 3 s, respectively.
The final solution was set aside for 10 min before being passed
through the mixed cellulose ester membrane filter. After filtration, 6 mL 0.1M HCl was used to wash out the free BPB adsorbed on the filter. Washing solution used at a volume larger
than 10 mL would give lower recovery.

Results and Discussion

Optimum Conditions for Spectrophotometric

Determination of CS+
Selection of anionic dye is very important in this method
because the CS+ is determined by measuring the absorbance
of anionic dye which is present at the same concentration as
the CS+ in the DMF. The anionic dye should give quantitative
recovery of CS+, good selectivity, higher molar absorptivity,
and lower reagent blank. In our previous work (15), we examined several anionic dyes, and proposed MO as the counter ion
for determination of CS+ in water. For the sediment sample,
however, MO was not feasible. The chlorophyl which was
co-extracted with CS+ from sediment samples has a maximum
absorption at 425 nm, quite near the absorption maximum of
423 nm for MO. In this work, BPB (Figure 2) was an ideal anionic dye that effectively avoided interference from
chlorophyl (Figure 3). The basic form of BPB reaches its

Figure 3. Absorption spectra of substance extracted

from sediment, the basic form of MO and BPB in DMF:
(a) substance (mostly chlorophyl) obtained by following
procedures for sediment sample determination but
without addition of anionic dye; (b) acid form of BPB in
DMF, 6.0 105Mol/L; (c) MO in DMF, 6.0 105Mol/L;
(d) basic form of BPB in DMF, 6.0 105Mol/L.

1784 HUI-FENG ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 84, NO. 6, 2001
Table 1. Recoveries of CS+ in water samples
Zephiramine
added, M CS+ found,a M Recovery, %

Water
Ground water

Jinzu River

Yonmangoku River
c

Furu River

a
b
c

NDb

0.30

0.30

100

ND

0.30

0.29

97

ND

0.30

0.28

93

0.07

0.30

0.35

93

such as Cl, SO42, NO3, and HPO42 on the determination

was investigated. These ions, at the concentration about
3 times higher than the average concentrations of those in the
actual river waters, did not seriously interfere with the determination of 0.3 M Zephiramine. Recoveries of Zephiramine
added to ground water and river water were >93% (Table 1).
In the Furu River water, which contains domestic waste, CS+
was detected at 0.07 M.

Examination of Sediment Samples

Mean of 3 determinations, equivalent to Zephiramine.

Less than 0.02 M.
Containing domestic waste water.

Validation of Extraction and Preconcentration

Procedures

Examination of Water Samples

A PTFE membrane filter with 0.1 m pore size was used to
extract CS+-AS ion associate. PTFE membrane is an effective extraction medium for ion associates (21). Futhermore, it
has very high solvent resistance and can be reused more than
10 times. A 5 mL volume of distilled water was applied to
clean the PTFE membrane. The CS+-AS ion associate collected on the PTFE membrane could be completely eluted
with only 2 mL methanol. The AS in the methanol solution
equaled the amount of CS+ and did not interfere with the formation of CS+-BPB in the following step. A linear calibration
curve was obtained for Zephiramine in the range of
0.0750.6 M, with a detection limit of 0.02 M, based on
3 times the standard deviation of the reagent blank. Relative
standard deviations for Zephiramine at concentrations
>0.15 M were <5.0%.
The effect of pH on water samples was examined. The
absorbance remained constant within the pH range studied. In
this work, pH of the water samples was adjusted to 2 to minimize the adsorption of CS+ onto the walls of glass vessels after
sampling. The effect of various common cationic ions such as
K+, Ca2+, Mg2+, Al3+, and NH4+, and the common anionic ions
+

Table 2. Effect of extraction times on extraction of CS

from sediment samplea

Methanol was used as the solvent for the ultrasonic extraction of CS+ in sediment. The output power and the frequency
of the ultrasonic generator were 120 W and 58 kHz, respectively. Concentration of CS+ in methanol solution was constant when irradiation time exceeded 90 min. However, CS+
could still be detected after the sediment was added to the new
portion of methanol and once again irradiated, indicating that
the CS+ could not be completely extracted from the sediment
with just one extraction operation. In this work, CS+ in a sediment sample was extracted 3 times, with each irradiation time
lasting 60 min. The CS+ in each methanol solution was determined separately. Total extractions were calculated from the
summation of the absorbance for each extraction time (Table 2). Nearly 96% extraction was obtained after 2 extractions.
Other organic substances in sediment, such as chlorophyl,
were also extracted into the methanol solution together with
the CS+. A recovery test was conducted by adding 1.5 and
3.0 M Zephiramine to the filtered methanol solution. The recovery was >95%, indicating that the organic substances did
not interfere with the spectrophotometric determination.

Investigation for Other CS+s

The other CS+s such as alkyltrimethylammonium and
benzethonium salts, which are commonly used in many fields,
were also investigated. The absorbances for benzethonium
were about 90% of that for Zephiramine at the same concentration. Distearyldimethylammonium ion gave an absorbance
15% lower than Zephiramine. Trimethylstearylammonium,
cetyltrimethylammonium, and benzyldimethylstearylammonium
ions gave about 80% of that for Zephiramine. For cetylpyridinium,
Table 3. Concentrations of CS+ detected in river water
and sediment samples
CS+ found in water,
mol/1000 g

CS+ found in sediment,

mol/1000 g

0.06

2.8

0.07

25.5

0.02

5.4

12.8

Sampling site
Place A

Extraction times
1

Place B
3

Place C
Place D

Absorbance
Total extraction, %
a

0.676
80.1

0.131
95.6

ND

0.037
100.0b

An aliquot of 50 g dried sediment sample was used. Ultrasonic

irradiation time for each extraction was 60 min.
Total extraction was supposed to be 100% after 3 extractions.

a
b
c
d
e

60 m upstream from place B.

A discharging point of domestic waste water.
80 m downstream from place B.
400 m downstream from place B.
Less than 0.02 mol/1000 g water.

HUI-FENG ET AL.: JOURNAL OF AOAC INTERNATIONAL VOL. 84, NO. 6, 2001 1785

the absorbance was 30% lower than that for Zephiramine. The results could be explained by the difference between the hydrophobic properties of the CS+s which affected the formation of ion associates and the extraction to membrane filters.

Behavior of CS+ in Aquatic Environments

The proposed method was applied to the determination of
CS+ in both river water and sediment. The Furu River was selected for that purpose because it contains domestic waste water. The water and sediment were sampled at 4 places (AD)
on January 19, 1999. Place B was a discharging point of domestic waste water. Place A was 60 m upstream from the discharging point, places C and D were 80 and 400 m downstream from the discharging point, respectively (Table 3).
Compared with the extremely low concentration of CS+ in water samples, some amounts of CS+ were detected in all
4 places. Furthermore, the concentration of CS+ in sediment
from place B was much higher than that from places either upstream or downstream. It was clear that the CS+ was quite easily adsorbed on the sediment surface by means of cationic exchange or hydrophobic interaction, and only a very small
portion of CS+ existed in the river water.
Conclusions
A simple method was developed for the determination of
CS+ in river water and sediment with 2 membrane filters, solvent soluble and insoluble. For water samples, it effectively
avoids interference from the co-existing AS, as described in
other methods, and requires only methanol rather than the
highly toxic solvent used in the solvent extraction method. For
sediment samples, it could be the first approach to the determination of CS+. In the river containing domestic waste water,
the concentration of CS+ in both cases was determined. Results indicated that CS+ was quite easily adsorbed onto the surface of river sediment.
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