ANALYTICAL SCIENCES 2001, VOL.

17 SUPPLEMENT i1329
2001 © The Japan Society for Analytical Chemistry

Homogeneous Liquid-liquid Extraction as Preconcentration Method

Using Water / Tetrabutylammonium Bromide / Chloroform Ternary
Component System and Its Application to the Flame-less Atomic
Absorption Spectrometry

Shigekatu OSHITE†1 and Shukuro IGARASHI 2

1†
Department of Chemistry and Biochemistry, Fukushima National College of Technology, 30 Nagao,
Kamiarakawa, Taira, Iwaki-shi, Fukushima, 970-8034, Japan (E-mail: soshite@fukushima-nct.ac.jp)
2
Department of Materials Science, Faculty of Engineering, Ibaraki University, Nakanarusawa, Hitachi,
Ibaraki, 316-8511, Japan

The new homogenous liquid-liquid extraction method based on pH independent phase separation phenomenon was
established using a water/tetrabutylammonium ion (TBA+)/chloroform ternary solvent system. This phase separation
phenomenon occurred by an ion-pair formation of TBA+ and perchlorate ion. A concentration factor of 260 (i.e. 39 cm3
→150 µl) was achieved by adjusting the initial volume of chloroform. In the extraction of Fe(Ⅱ)-phenanthroline complex,
the distribution ratio and the extraction percentage were 1.1×103 and 80 %, respectively. Satisfactory results for the
flame-less atomic absorption spectrometric determination of iron(II) was obtained by the combination of this method and 1,
10-phenanthroline complex formation {linear range: 0.060-125 ppb for Fe(II)}.

(Received on August 9, 2001; Accepted on September 13, 2001)

Homogeneous liquid-liquid extraction method is which extracts
the desired solute existing in the homogeneous aqueous solution
into the water-immisible sedimented phase based on each kind of
phase separation phenomena. The methods, which used the
phase separation depended upon the temperature of the
water/propylene carbonate system 1 or based on the salt effect in
the water/acetonitrile system, etc., have been already reported 2, 3.
Moreover, these methods have merit in which the extraction
speed becomes extremely fast due to the absence of obstacles on
contact surface between the aqueous phase and organic phase
during the extraction procedure.
Recently, homogeneous liquid-liquid extraction method using
a ternary solvent system has been developed 4 and it was applied
as a simple and high-powered preconcentration for the
instrumental analysis (e.g. VIS spectrophotometry 5, HPLC 6,
and GC/MC 7). In concretely, the methods based on the pH
dependent phase separation, which used a water/acetic
acid/chloroform ternary solvent system5 or
water/pyridine/ethyl chloroacetate system8, were reported.
However, in these methods, the widespread application for target
analytes has been limited by the inconvenient pH condition in
extraction. In this study, a new pH independent phase separation
phenomenon was researched using the ion-pair formation of
tetrabutylammonium ion (TBA+) and perchlorate ion (ClO4-) in
homogeneous solution (water/TBA+/chloroform). The
homogeneous liquid-liquid extraction method based on this
phenomenon, which is able to simply concentrate the Fe(II)-1,
10-phenanthroline complex into a microvolume of chloroform
phase, was developed. Moreover, this proposed method was
applied to the flame-less atomic absorption spectrometric
determination of small amounts of Fe(II). Details will be
discussed in this paper.
Experimental
Materials and apparatus
The quaternary ammonium bromides were used the both of
tetrabutylammonium bromide (Tokyo Kasei Co.) and n-
octyltrimethylammonium bromide (Kanto Chemicals). These
were the high-purity grade reagent. The chloroform, 4-methyl-
2-pentanone (MIBK) and sodium perchlorate were received from
Kanto Chemicals. All reagents used were of analytical reagent
grade, unless otherwise described.
A Nippon Bunko V-560 model UV-VIS Spectrophotometer
and a Hitachi Z-8000 model polarized zeeman atomic absorption
spectrophotometer with graphite tube cuvette was used.
Standard procedure for the examination of phase separation
a phenomenon
A homogeneous solution (water/TBA+/chloroform) was
placed in a 50cm3 cylindrical glass vial fitted with a plastic cap.
TBA+ aqueous solution was added and the mixture gently shook.
The mixture was allowed to stand for 10 min at 25°C in a
thermostated bath. After adding NaClO4 aqueous solution to the
mixture, it was centrifuged for 5 min at 1800 rpm. The volume
of sedimented phase was determined using a 100 µl micro-
syringe. The concentration of the solute in the aqueous phase
and/or the water-immisible sedimented phase was
spectrophotometrically determined.
i1330
Procedure for the determination of Fe(II)
Based on phenanthroline complex formation9, a 30 cm3 of the
sample solution containing Fe(II) was complexed and it was
extracted using the proposed method. The concentration of
Fe(II) in the sedeminted chloroform phase was determined by
injection its phase into graphite tube cuvette of an atomic
absorption spectrometry (the carrier gas; 200 cm3/min,
interraputed gas; 30 cm3/min, the volume of sample; 30µl, the
detection wavelength; 248.3 nm, the dry; 60-120 °C (30s), the
aching; 600 °C (30s), the atomization; 2700 °C (10s) and the
clean; 2800 °C (3s)).
Results and Discussion
Mechanism of phase separation
In the already reported methods, the mechanism of phase
separation was postulated 5, 8. When the organic solvent such as
chloroform and ethyl chloroacetate is respectively solvated by
the molecule such as acetic acid and pyridine, the organic
solvent dissolves in water. However, when the solvation effect
of the molecule is excluded by the change of pH, the water-
immisible sedimented phase which consists of organic solvent in
the aqueous solution is produced by the phase separation. In this
proposed method, when NaClO4 is added to the homogeneous
solution which consists of a water/TBA+/chloroform, the
solvation effect of TBA+ is excluded by the ion-pair formation of
TBA+ and ClO4-. Therefore, the chloroform in homogeneous
aqueous solution is postulated to form water-immiscible
chloroform in aqueous solution by the phase separation.
Selection of quaternary ammonium ion (Q+) and organic solvent
in phase separation
Two kinds of the quaternary ammonium ion (Q+) with a
different form of a molecule were used to increase the solubility
of chloroform to water (i.e., TBA+ with the total carbon number
of 16 and octyltrimethylammonium ion (OTMA) with 11). The
ClO4- was selected for the counter ion of Q+, because it is usually
used for the reagent of an ion-pair extraction. As a result, the
phase separation phenomenon in ternary solvent system was
observed using not only TBA+ but also OTMA+. However, the
added ClO4- aqueous solution on water/OTMA+/chloroform
system needed higher concentration than that on
water/TBA+/chloroform system, because the solubility of
OTMA+ · ClO4- to water is higher than that of TBA+ · ClO4-.
Moreover, two kinds of the organic solvent were selected (i.e.
chloroform as usual organic solvent, and MIBK as the solvent of
atomic absorption analysis). The phase separation phenomena in
ternary solvent system were observed using not only chloroform
but also MIBK. However, the chloroform was selected in order
to run the experiment easily, because the separeted MIBK phase
floated on water (the specific gravity of MIBK: 0.8). Hence,
TBA+, chloroform and NaClO4 were selected.
Optimum conditions of phase separation in ternary solvent
system
The conditions, which produce a homogeneous phase or two
phases, were examined by using different compositions of water,
TBA+ and chloroform (Fig. 1, curve a). Moreover, NaClO4
aqueous solution was added in order to form ion-pair of TBA+
and ClO4-, and the existence of the phase separation from a
homogeneous solution was examined (Fig. 1, curve a-b). As a
result, the optimum composition conditions for each solvent in
the homogeneous liquid-liquid extraction were obtained (marked
with a symbol × in Fig. 1: [CHCl3]T = 0.74 %, [TBA]T = 2.0 ×
10-2 mol dm-3, [ClO4-]T = 2.6 ×10-3 mol dm-3). For determining
these conditions, the concentration of NaClO4 and chloroform
need to be as low as possible, and the stable sedimented phase
was considered.
ANALYTICAL SCIENCES 2001, VOL.17 SUPPLEMENT
a
1 .0
C H C l 3 / v ol%
b
0 .6
0 .2
0 0 .5 1 1 .5 2 2 .5
+ -3
[T B A ] T / m o l d m
Fig. 1 Phase separation diagram in water/TBA+/chloroform
ternary solvent system using two kinds of concentration for
sodium perchlorate at 25°C. Various total concentrations of
sodium perchlorate were added: curve a: 0, b: 2.6×10-3mol dm-3.
The upper region of each curve shows the state of two phases
and the lower region shows the homogeneous phase.
Concentration of ClO4-
The relationship between the concentration of NaClO4 and the
volume of the sedimeted phase is shown in Fig. 2. The phase
separation phenomenon based on ion-pair formation of TBA+
and ClO4- was observed in the ClO4- concentration range of 1.3
×10-2 -2.5 ×10-2 mol dm-3. The volume of sedimented phase
was constant (i.e. 150 µl),.because the composition of ion-pair
becomes approximately 1:1 in the neighborhood of [TBA+]T =
[ClO4-]T. Also, the salting-out effect clearly appeared above the
high concentration of NaClO4 2.5 ×10-2 mol dm-3. As a result,
an increase in the volume of sedimented chloroform phase was
observed. In this system, Fe(II)-phenanthroline complex was
selected as a cationic solute model. Also, the adding amount of
NaClO4 was determined by the extraction percentage. Namely,
the distribution ratio (D) and the extraction percentages (E%)
were respectively 1.1 × 103 and 80 % in the condition of
[TBA+]T < [ClO4-]T, but they were 1.4×102, and 36 % in the
condition of [TBA+]T > [ClO4-]T. In former condition, the
complex was extracted to chloroform phase by the ion-pair
formation of the complex and the excess amount of ClO4-.
Therefore, the optimum concentration condition of NaClO4 was
2.3 ×10-2 mol dm-3.
3 00
V o lu m e of se dim e nte d p has e / µl
2 00
1 00
0
0 1 2 3
[ N aC lO 4 ] T / 10 -2
m ol dm -3
Fig. 2 The relationship between sodium perchlorate
concentration of sedimented phase. [TBA+]T = 2.0×10-2 mol
dm-3; [CHCl3]T = 0.74 vol%; 25°C; total volume: 39 cm3.
ANALYTICAL SCIENCES 2001, VOL.17 SUPPLEMENT i1331

200 Homogeneous liquid-liquid extraction of Fe(II)-phenanthroline

complex
V o lum e o f se dim en te d p hase / µl

Free Fe(II) ion and Fe(II)-phenanthroline complex were

selected as a cationic solute model and they were
spectrophotometrically examined. In the extraction of Fe(II)
compounds, when the volume ratio (Va/Vs) was 260, the D and
E% were 15 and 5 % for Fe(II) and 1.1×103 and 80 % for
100 Fe(II)-phenanthroline complex, respectively. This high value of
E% was obtained because of high hydrophobicity of the ion-pair
compound between the complex and ClO4-.

Application to flame-less atomic absorption spectrometry

The flame-less atomic absorption spectrometric determination
of iron(II) with the combination of 1,10-phenanthroline complex
0 formation and the proposed method was developed. The
0 100 200 300
calibration graph was linear over the concentration range of
V o lu m e o f ad d ed ch lo ro fo rm / µ l 0.060-125 ppb.

Fig. 3 The relationship between the volume of added chloroform The pH independent phase separation phenomenon was found
and the volume of sedimented phase. [TBA+]T = 2.0×10-2 mol using the ion-pair formation of TBA+ and ClO4-. A proposed
dm-3; [NaClO4]T = 2.3×10-2 mol dm-3; at 25°C. method based on this phase separation phenomenon could
separate and concentrate the target analyte by a simple procedure.
Microvolume of the sedimented chloroform phase Moreover, this method was able to apply to the flame-less
The relationship between the volume of added chloroform and atomic absorption spectrometric determination of small amounts
the volume of the sedimented phase is shown in Fig. 3. As a (several 10 ppt level) of iron(II). In the near furture, the
result, the volume of sedimented phase (i.e. 150 µl) was constant proposed method will be widely applied as simple and high-
in the volume range of added chloroform 180-280 µl. When the powered preconcentration of practical instrumental analyses
volume of added chloroform was less than 150 µl, the solid state beginning with AAS.
of the sedimented phase was produced to excess the solubility of
an ion-pair to chloroform. Moreover, the curve in figure did not
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200

100

0
1 2 3 4 5 6 7 8 9
pH

Fig. 4 The relationship between pH and the volume of

sedimented phase. [TBA+]T = 2.0×10-2 mol dm-3; [NaClO4]T =
2.3×10-2 mol dm-3; at 25°C