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Separasi Arsenik
Separasi Arsenik
@ Springer-Verlag 1992
Summary. Capillary zone electrophoresis (CZE) in capillary resis (FSCE) is based on differences in the electrophoretic
silica columns has been used for the separation of arsenite mobilities of species. These parameters depend on the
(AsOj), arsenate (AsO~-), monomethylarsonic acid structural differences of the solutes (size, shape and charge)
(MMA) and dimethylarsinic acid (DMA). The separation and are also affected by the physico-chemical properties of
of these ionic species has been achieved using a capillary the carrier electrolyte, such as pH, ionic strength, viscosity
silica column (72 cm x 50 gm i.d.) with an acidic phosphate and dielectric constant. Many ions, such as iodate and
buffer and with an on-column UV detection (190 nm). periodate [23], metal ions [24] and hexacyanoferrate(II) and
Optimization of experimental parameters (pH, temperature, -(III) ions [25] can be separated by FSCE.
voltage) were studied. The selectivity of the separation can In this paper, the separation of arsenic species by
be improved by working in the pH-range of 4 . 5 - 6.5. For capillary electrophoresis with an on-column UV detection is
analytical inorganic separations of UV-absorbing anions, presented.
capillary zone electrophoresis has advantages because of the
relatively simple equipment, the short analysis time (15 min),
the high efficiency and the low mass detection limit (40 pg Experimental
for arsenate). Capillary electrophoresis. An Applied Biosystems (Foster
City, CA, USA) Model 270A capillary electrophoresis
system with a fused silica column was used for this work. A
72 cm × 50 ~m i.d. silica capillary filled with phosphate buf-
fer served as the separation tube operated at high voltage
Introduction (10-40 kV). A small section of the polyimide coating of the
capillary column was removed prior to filling to get an
Arsenic is present in both marine and freshwater organisms optical window for UV detection. Between analyses, the
in the form of water-soluble and lipid-soluble compounds capillary was flushed with the buffer solution for 15 min.
[1]. The level of total As in sea water is approximately 2 ~g/ The on-column detection was operated at 190 nm at an
kg [2, 3]. Four different arsenic species, arsenite [As(III)], absorbance range of 0.01 AUFS and a rise time of 1 s. The
arsenate [As(V)], monomethylarsonic acid (MMA) and di- column temperature was set at 40 ° C. Samples were injected
methylarsinic acid (DMA), have been detected in sea water by the vacuum technique. All electropherograms were re-
[4]. Analysis of marine algae has shown that arsenic is present corded using a Spectra Physics (San Jos6, CA, USA)
in other forms than inorganic or methylarsenic compounds Chromjet Model integrator.
[5]. Several papers describe the speciation analysis of arsenic
by ion chromatography [6-11] or reversed-phase liquid Chemicals. Commercially (Prolabo, France) available
chromatography [12-17]. An alternative analytical method sodium arsenite NaAsO2 (purity 99%), sodium arsenate
is capillary electrophoresis in which the analyte is introduced Na2HAsO¢- 7 H20 (purity 98.5%), sodium dimethylarsinate
into a capillary tube and subjected to eleetrokinetic separa- (DMA) or sodium cacodylate (purity 98%) were used
tion. Extensive reviews of capillary electrophoresis have been without further purification. The synthesis, purification and
recently published in different journals [18-22]. Capillary characterization of sodium methylarsonate (MMA) has been
electrophoresis (CE) is a very powerful separation technique achieved in the "Service Central de Microanalyse" (Dr. A.
for the separation of ionic as well as neutral substances. Lamotte), CNRS, Vernaison (France).
Narrow diameter capillary (50-75 ~Lm i.d.) leads to an Water used for dilutions or as buffer solution was HPLC
efficient dissipation of heat and suppresses convection, grade (Rathburn). The phosphate buffer solution was pre-
whereas the high voltage causes fast separations with high pared by mixing in various proportions stock solutions of
efficiency (10 s -106 theoretical plates within 30 min). The 0.067 tool/1NaH2PO4 and 0.067 mol/l NazHPO4 (Prolabo).
separation mechanism in free solution capillary electropho- Stock solutions of arsenical species (100 ppm) were prepared
in HPLC grade water and kept at - 2 0 ° C as a precaution
Offprint requests to: M. Leroy against any degradation.
358
[ I I ~ I I I I
-3.5 Effect of p H on electroosmosis
2 4 6 8 10 12 14 16
pH In order to determine the electrophoretic mobility of a solute
by CZE, we must first consider the influence of electro-
Fig. 1. Variation of the calculated charge of several arsenic ionic endosmosis of the measurements. The apparent mobility ]-~app
species versus pH. Solutes: 1 arsenite; 2 dimethylarsenate; 3 mono- is defined as:
methylarsonate; 4 arsenate
[~app = Ld" LT/t' V; (2)
where Ld is the length of the capillary from the inlet to the
Resultsanddiscussion detector, LT the total length of the capillary, t the migration
time and V the applied voltage•
Variation of the charge of arsenic species versus p H In CZE, the migration time (to) for a neutral marker is
defined as:
The arsenic compounds contain acidic groups in their
chemical structure, so their apparent charge depends on to = La" LT/Ueo " V ; (3)
359
3 ~epX 104(cm2/V.s)
6 4
0,01 AU
1
0,01 AU
2
0 I I ~ J J
I 4 5 6 7 8 9
pH
I Fig. 3. Effect of pH on the electrophoretic mobilities of arsenic
species. Silica capillary: 72 cm x 50 gm i.d.; carrier: phosphate buf-
i
r
w
e4
=_.
. I 0.01 AU 0.01 An
m 00" ~n !
i
.0
[
i~,,d
c
0
Time ( r a i n ) - ,
(a)
10 0
-Time (min),,,
(b)
]
]
I'0 0
~T£me
J(rain)--v
(c)
!0
C Time (rain)-~
(d)
to
~ T i m e (min I'-'7
0
{e)
10 0
r.---Time
{f)
W
(rain)--q
10
Fig. 4 a - L Effect of buffer pH on the selectivity of arsenic species by FSCE. Silica capillary: 72 cm x 50 lam i.d.; carrier: phosphate buffer
25 mmol/1; applied voltage: 30 kV; temperature: 40°C; detection by UV absorption at 190 nm; mixture concentration was 100 gg • m l - ~
of each species; injected volume: 9 nl (900 pg of each species). Solutes: 1 arsenite; 2 dimethylarsenate; 3 monomethylarsonate; 4 arsenate.
a pH 5.00; b pH 5.29; c pH 6.24; d pH 6.64; e pH 6.81; f pH 7.17
(a) (h)
450
4
400
350
300
2O0
150
2
100
50 I I t l I I --I " t
24 26 28 30 32 34 86 38 40
T ¢c)
Fig. 7. Dependence of the electrophoretic mobilities of arsenic species on the temperature. Silica capillary: 72 cm x 50 gm i.d. ; carrier:
phosphate buffer 25 mmol/l, pH 5.6; applied voltage: 30 kV; detection by UV absorption at 190 nm; mixture concentration was 100 gg •
ml -~ of each species; injected volume: 9 nl (900 pg of each species). Solutes: I arsenite; 2 dimethylarsenate; 3 monomethylarsonate;
4 arsenate
consequently, an interesting selectivity happens for a buffer pherogram (Fig. 4f). The same phenomenon happened for
pH close to 6.5 value, but the resolution always remains M M A above p H 8.2.
important in the p H range 4 . 5 - 6.8. Under these conditions (phosphate buffer 25 mmol/1, p H
Arsenate and then M M A disappear from the electro- 5.6), arsenite (tr = 2.5 min), D M A (t~ = 2.85 rain), M M A
pherogram at higher pH-values. At p H 7.17, the velocity of (tr = 4.96 rain), arsenate (tr = 5.54 rain) gave sharp and
electrophoretic migration for arsenate was so high that this symmetrical peaks well resolved from each others, as shown
anion (Qapp = --1-65), introduced to the anodic end of in Fig. 5. The number of theoretical plates was rather high
the silica capillary, was forced back into the anodic buffer; [132000 for As(IID], one order or more higher than the
consequently, only three peaks appear on the electro- usually encountered values in HPLC. The selectivity allows
362