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A contactless capacitively coupled conductivity detector Another possibility to specifically detect ionic species with a
for capillary electrophoresis is introduced. The detector high equivalent conductivity is to measure the differences in
consists of two electrodes which are placed cylindrically conductivity of the solute zones and the separation electrolyte.
around the outer polyimide coating of the fused-silica By using separation electrolytes with a low conductivity, a sensitive
capillary with a detection gap of 2 mm. The electrodes detection is thus possible. Several devices have been described
form a cylindrical capacitor, and the electric conductivity that use direct conductivity detection for cations and anions. The
of the solution in the gap between the electrodes is detection can be accomplished either by a direct contact of the
measured. A high audio or low ultrasonic frequency for solution with the electrodes19-26 or by contactless methods which
coupling of the ac voltage is used in order to minimize require inductively or capacitively coupled devices using high
the influence of reactance of the liquid. For an improved frequencies.27-35 Most of these instruments were originally
version of the detector, two syringe cannulas are used as intended to work for chromatographic and isotachophoretic
the electrodes and the capillary is simply assembled into purposes; however, on principle they could be used for capillary
the tubing. This allows an easy placement of the detector electrophoresis as well.
on various positions along the capillary. The limit of A certain drawback of instruments working with conductivity
detection of inorganic cations and anions is 200 ppb, as detection measured by direct contact of electrodes and sample
determined for sodium and chloride, respectively. are the high instrument costs, as the capillary has to be modified
to a specific extent and a rather complicated geometry and
Efficient on-column detection for capillary electrophoresis is
(10) Bächmann, K.; Boden, J.; Haumann, I. J. Chromatogr. 1992, 626, 259.
aggravated by the small volumes when capillaries with inner (11) Xue, Y.; Yeung, E. S. Anal. Chem. 1993, 65, 2923.
diameters between 25 and 75 µm are used. However, ionic species (12) Shamsi, S. A.; Danielson, N. D. Anal. Chem. 1994, 66, 3757.
(13) Buchberger, W.; Cousins, S. M.; Haddad, P. R. Trends Anal. Chem. 1994,
in the injection plug are being concentrated by electrostacking
13, 313.
while forming distinctively separated and narrow zones when an (14) Shamsi, S. A.; Danielson, N. D. Anal. Chem. 1995, 67, 4210.
electric field is applied. This effect also contributes to the high (15) Salimi-Moosavi, H.; Cassidy, R. M. Anal. Chem. 1995, 67, 1067.
(16) Desbene, P. L.; Morin, C. J.; Monvernay, A. M. D.; Groult, R. S. J.
separation efficiencies of the method. As a consequence, the
Chromatogr., A 1995, 689, 135.
concentration of the solutes in the respective band zones can be (17) Collet, J.; Gareil, P. J. Chromatogr., A 1995, 716, 115.
many times higher as compared to the original injection plug, (18) Weston, A.; Brown, P. R.; Jandik, P.; Heckenberg, A. L.; Jones, W. R. J.
Chromatogr. 1992, 608, 395.
which significantly facilitates on-column detection.1
(19) Huang, X.; Pang, T. K. J.; Gordon, M. J.; Zare, R. N. Anal. Chem. 1987, 59,
The most widely used mode of detection is UV absorptivity 2747.
detection, which requires the presence of chromophoric moieties. (20) Harrold, M.; Stillian, J.; Bao, L.; Rocklin, R.; Avdalovic, N. J. Chromatogr., A
1995, 717, 371.
However, inorganic cations and anions do not exhibit a sufficiently
(21) Avdalovic, N.; Pohl, C. A.; Rocklin, R.; Stillian, J. Anal. Chem. 1993, 65,
high absorptivity to be detected by UV absorption at low 1470.
concentration levels. As a consequence, indirect photometric and (22) Huang, X.; Luckey, J. A.; Gordon, M. J.; Zare, R. N. Anal. Chem. 1989, 61,
766.
fluorometric UV detection modes have been introduced.2-18
(23) Huang, X.; Zare, R. N.; Sloss, S.; Ewing, A. G. Anal. Chem. 1991, 63, 189.
(24) Huang, X.; Zare, R. N. Anal. Chem. 1991, 63, 2193.
* Corresponding author: (Tel) +43-512-507-5180; (Fax) +43-512-507- (25) Avdalovic, N.; Pohl, C. A.; Rocklin, R. D.; Stillian, J. R. Anal. Chem. 1993,
2965; (e-mail) andreas.zemann@uibk.ac.at. 65, 1470.
(1) Jandik, P.; Bonn, G. Capillary Electrophoresis of Small Molecules and Ions; (26) Haber, C.; Jones, W. R.; Soglia, J.; Surve, M. A.; McGlynn, M.; Caplan, A.;
VCH Publishers: New York, 1993; p 118ff. Reineck, J. R.; Krstanovic, C. J. Capillary Electrophor. 1996, 3, 1.
(2) Yeung, E. S. Acc. Chem. Res. 1989, 22, 125. (27) Alder, J. F.; Drew, P. K. P. Anal. Chim. Acta 1979, 110, 325.
(3) Kuhr, W. G.; Yeung, E. S. Anal. Chem. 1988, 60, 2642. (28) Müller, D.; Jelinek, I.; Opekar, F.; Stulik, K. Electroanalysis 1996, 8, 722.
(4) Gross, L.; Yeung, E. S. J. Chromatogr. 1989, 480, 169. (29) Pal, F.; Pungor, E.; Kovats, E. Anal. Chem. 1988, 60, 2254.
(5) Foret, F.; Fanali, S.; Ossicini, L.; Bocek, P. J. Chromatogr. 1989, 470, 299. (30) Alder, J. F.; Drew, P. K. P. Anal. Chim. Acta 1979, 110, 325.
(6) Yeung, E. S.; Kuhr, W. G. Anal. Chem. 1991, 63, 275A. (31) Gas, B.; Demjanenko, M.; Vacik, J. J. Chromatogr. 1980, 192, 253.
(7) Vorndran, A. E.; Oefner, P. J.; Scherz, H.; Bonn, G. K. Chromatographia (32) Everaerts, F. M.; Rommers, P. J. J. Chromatogr. 1974, 91, 809.
1992, 33, 163. (33) Vacik, J.; Zuska, J.; Muselasova, I. J. Chromatogr. 1985, 320, 233.
(8) Beck, W.; Engelhardt, H. Chromatographia 1992, 33, 313. (34) Decristoforo, G. Thesis, University of Innsbruck, 1979.
(9) Bächmann, K.; Haumann, I.; Groh, T. Fresenius J. Anal. Chem. 1992, 343, (35) Schnell, E.; Zemann, A. J.; Volgger, D.; Bonn, G. K. Patent pending (No.
901. A1016/97), 1997.
S0003-2700(97)00759-2 CCC: $15.00 © 1998 American Chemical Society Analytical Chemistry, Vol. 70, No. 3, February 1, 1998 563
Published on Web 02/01/1998
construction is required. Furthermore, changes in the cell
geometry can be considered disadvantageously.28 As a conse-
quence, for capillary electrophoresis purposes, a detector without
galvanic connection of solutes and electrodes with, at the same
time, equal sensitivity as compared to a directly connected
conductivity detector is a certain demand.
An oscillometric detector for ion chromatography has already
been described with electrodes being separated from the elec-
trolyte by a silicon lacquer or Teflon coating.29 This device enables
a better reproducibility and stability than detectors with a galvanic
contact. Generally, with detectors using a high-frequency volt-
age29,31,34 a signal resulting from both ohmic resistance and
reactance of the liquid is recorded. Inductively coupled contact-
less conductivity detectors have been described as well.27,30
Figure 1. Schematic drawing of the capacitively coupled conductiv-
Several attempts have been made to use contactless conductiv-
ity detector (CCCD). The electrodes are made of either a conducting
ity detection for capillary isotachophoretic purposes. Instruments silver varnish or of metallic syringe cannulas with the capillary
consisting of an arrangement of four electrodes placed outside assembled through.
the PTFE capillary have been described which use a high-
frequency voltage.32,33 Polarization of the measuring electrodes
or undesirable redox reactions are thus prevented. The applica- to the power supply, an RFI line filter was used in order to avoid
tion of conductivity detection in isotachophoresis is described in noise from impulses and spikes.
detail in various books on isotachophoresis.36-38
The present paper describes the application of a capacitively RESULTS AND DISCUSSION
coupled conductivity detection system (CCCD) which works on The conductivity detector described in this paper works on a
a contactless basis for the detection of cationic and anionic contactless basis. It is used in connection with a commercial
compounds after capillary electrophoretic separation. capillary electrophoresis instrument (Waters Quanta 4000) which
enables free handling and accessibility of the capillary along almost
its complete length. Among other advantages, this system enables
EXPERIMENTAL SECTION
an arbitrary placement of the detector as no capillary cartridge is
Instrumentation. A Quanta 4000 capillary electrophoresis
used. Virtually any effective separation length can be adjusted
system (Waters Chromatography, Milford, MA) connected with
without the need of removing the polyimide coating which is
an AD changer (System Interface Module, Waters) and a personal
necessary for on-column UV detection. Thus, fragile detection
computer was used. Data acquisition and processing was carried
windows are avoided and the risk of breaking the capillary during
out with a commercial chromatography software (Maxima 820,
handling is reduced.
Waters). Fused-silica capillaries (Composite Metal Services,
Figure 1 depicts a schematic drawing of the CCCD. Two
Worchester, U.K.) with an inner diameter of 50 µm and an outer
cylindrical conducting surfaces (electrodes) are placed around the
diameter of 375 µm were used. Sample injection was carried out
polyimide coating on the outside of an uncoated fused-silica
hydrostatically at an elevation of 10 cm.
capillary. The electrodes can be made either of a conducting silver
Reagents. All reagents were of analytical grade (Sigma-
varnish which is painted around the polyimide coating of the
Aldrich Handels-Ges.m.b.H, Vienna, Austria, and Merck GmbH,
capillary or of two syringe cannulas with the appropriate length
Vienna, Austria). Standard and buffer solutions were prepared
where the capillary is passed through.
by dissolving the respective compounds in ultrapure water
For experimental testing of the detector, 30 mm was chosen
(Barnstead/Thermolyne, Dubuque, IA).
as the length of each of the electrodes. This length can be varied
Electronic Parts. The contactless detection device was built
from 15 to 50 mm, which then, as a consequence, requires
from electronic components which were purchased from local
different amplification to obtain equal signal intensities. In fact,
electronic dealers. Two directly coupled transistors (BC 414),
changing the length of the electrodes has only little effect. An
which are connected to a potentiometer (100 kΩ) and an
increase of length to 50 mm renders only slightly differing
integrated circuit (U420B), were used for amplification. The
electropherograms in terms of sensitivity. For shorter electrodes,
rectification was performed using two diodes (BA4148). The
however, the respective amplification has to be increased which,
frequency generator that is suitable for a frequency of 40 kHz
at this time with the components used in this system, causes the
sinus was built from an LC oscillator circuit with a transistor (BC
signal-to-noise (S/N) ratio to become worse. The electrodes are
337). The 20-kHz rectangular form frequency generator was made
connected to an oscillator through a resistor with a resistance of
from an integrated circuit (Telefunken NE 555). The dc voltage
50 kΩ. Together with the reactance of the shielded cable
for the amplifier was stabilized to 9 V. For the mains connection
connection to the detector and the input resistance of the amplifier,
(36) Everaerts, F. M.; Beckers, J. L.; Verheggen, Th. P. E. M. Isotachophoresis: a resistance R of approximately 10 kΩ is formed. A voltage drop
Theory, Instrumentation and Application; J. Chromatogr. Library Vol. 6; of 0.2-2 mV is observed along resistance R, which is then
Elsevier: Amsterdam, 1976. amplified and rectified to direct current. The employed amplifier
(37) Everaerts, F. M. Analytical Isotachophoresis; Elsevier: Amsterdam, 1981.
(38) Bocek, P.; Deml, M.; Gebauer, P.; Dolnik, V. In Analytical Isotachophoresis; has a variable amplification of 500-1500-fold. After rectification,
Radola, B. J., Ed.; VCH Publishers: Weinheim-Basel, 1988. a direct voltage of 0.1-0.5 V is fed in the input of the AD changer
564 Analytical Chemistry, Vol. 70, No. 3, February 1, 1998
which itself is part of the data acquisition and processing system.
However, any conventional integrator or other AD changer can
be used for this purpose.
A detection gap between the two electrodes of 2 mm is chosen,
which can be increased to 5 mm, depending on the length of the
capillary and the required separation efficiency. A distance of
approximately 2 mm is sufficiently long with respect to important
separation parameters such as resolution and separation efficiency.
If the detection gap is too narrow, a capacitive transfer between
the electrodes may occur. As a consequence, if resolution
between specific solute zones is too low, it is advisable to increase
the capillary length instead of narrowing the detection gap.
The detection device described in this paper differs significantly
from other contactless conductivity detectors for isotachophoresis
or capillary electrophoresis purposes. Most of the other contact-
less conductivity detectors measure the signal radially across the
capillary, a principle that suffers from the very short path lengths.
Furthermore, the use of a low frequency bears several advantages
when compared to high-frequency conductivity detectors. In
addition to the ohmic resistance, a considerable dielectric con-
tribution of the reactance can also be assumed when high
frequencies are used. As a consequence, whenever the liquid
inside of one of the electrodes changes its dielectric properties,
which is likely when sample component zones are passing
through, a shift of the baseline would be detected before and after
the actual signal. Thus, comparable results with galvanically Figure 2. Separation of a standard mixture of inorganic cations
coupled detectors should be obtained in terms of relative signal comparing UV and conductivity detection. Conditions: fused-silica
intensities when low frequencies are used as primarily the ohmic capillary, L ) 60 cm, l1 ) 40 cm to conductivity detector, l2 ) 52.5
cm to UV detector (214 nm), U ) +20 kV, I ) 3.0 µA, and T ) 23
resistance along the detection gap is responsible for the signal. A
°C; electrolyte, 10 mM lactic acid, 8 mM 4-methylbenzylamine, and
frequency of 40 kHz sinus form or 20-30 kHz rectangular form 15% methanol, pH 4.9; injection, 30 s; sample concentration, 0.1 mM.
is used to keep the influence of reactance considerably low. With
the long cylindrical electrodes used in this investigation, this
frequency range works well with voltage amplitudes of 7-10 Vpp, the resistors are connected in series, this causes the voltage of
preferably with 8 Vpp. A higher oscillator voltage may improve all the other resistors to change significantly. As a consequence,
the S/N ratio to a certain extent, however, due to the limitations a voltage increase can be measured on resistance R. After
of the components and parts that build up the present detection amplification and rectification, the signal is fed into the AD
device, a significant improvement of the sensitivity is not achieved. changer.
The voltage drop on resistance R is below 1 mV. Thus, the The applicability of the developed device for capillary electro-
detector and all the connections have to be shielded to ground. phoretic purposes can be demonstrated for the detection of
In order to measure the differences of the conductivity between inorganic cations and anions. A comparison of the contactless
conductivity detection method with indirect UV detection provides
the electrolyte and the separated ions, an electric zero point
satisfactory results in terms of sensitivity as depicted in Figure 2.
depression is recommended as is found in any other instruments
The buffer consists of lactic acid, 4-methylbenzylamine, and
using direct conductivity with galvanically connected electrodes
methanol at a pH of 4.9. This system was originally introduced
or UV detection.
for indirect UV detection of cations.39 A 5-fold amplification of
The detector presents a series connection of two capacitors
the UV signal is necessary to achieve comparable peak heights
and two resistors. The electrodes serve as two independent
at given system settings.
cylindrical capacitors with the liquid inside the capillary being the
However, for practical purposes, a detector considered easy
corresponding internal wire. The detection gap between the
to use has to meet several basic requirements. Handling and
electrodes acts as one of the resistors and resistance R as the
mounting of the detection electrodes should be as easy as possible
other. A reactance between 2 and 4 MΩ at a frequency of 40
with only little pretreatment and modifications of the capillary.
kHz is calculated by assuming cylindrical capacitors. Using a
Although painting of silver conducting varnish on the outside of
buffer electrolyte with a low conductivity, the resistance of the
the capillary is both an inexpensive and easy pretreatment
liquid inside the capillary from the inlet to the outlet end is in the
procedure, the electrodes were subject to further improvement
range of several gigaohms. Thus, the electrolyte in the detection
of the detector. Two metallic syringe cannulas with an inner
gap between the electrodes, which measures approximately 0.3%
diameter slightly larger than the outside diameter of the silica
of the total capillary length, exhibits a resistance in the megaohm
capillary were cut to the appropriate length and used as electrodes.
range. If an ionic solute zone passes through the detection gap,
the resistance decreases due to the increasing conductivity. As (39) Shi, Y.; Fritz, J. S. J. Chromatogr. 1993, 640, 473.
CONCLUSION Received for review July 15, 1997. Accepted October 30,
Contactless capacitively coupled conductivity detection is a 1997.X
suitable detection method for inorganic anions and cations in
AC9707592
capillary electrophoresis. Placement of the capillary into two metal
tubings, which act as two cylindrical capacitors, allows a fast and X
Abstract published in Advance ACS Abstracts, December 15, 1997.