Electrophoresis 2014, 00, 1–6 1

Tomas Drevinskas1 Research Article

Audrius Maruška1
Vitalis Briedis2
Capacitance-to-digital: The upgrades of
1 Faculty of Natural Sciences,
Department of Biology,
single chip detector
Vytautas Magnus University,
Kaunas, Lithuania The capacitance-to-digital single chip detector was upgraded. The paper discusses hard-
2 Faculty of Pharmacy, Lithuanian
ware issues and benefits of the designed/upgraded detector. The device can be operated
University of Health Sciences,
Kaunas, Lithuania from rechargeable lithium-ion battery as stand-alone, portable system and is capable of
transmitting real-time data wirelessly. The detector and additional modules (battery, battery
holder, microcontroller board, wireless module) weight is less than 85 g. Electrophoretic
Received July 4, 2014 separation in low conductivity 20 mM MES/L-His buffer, pH 6.1, was performed in order
Revised September 11, 2014
to evaluate detection parameters. The system is capable of quantification of potassium ions
Accepted September 30, 2014
down to 0.31 ␮M. Investigation of differential signal acquisition configuration showed im-
proved performance regarding external noise and temperature fluctuations. The system
can be a solution for stand-alone, field-portable capillary format separation detector.

Keywords:
Capacitance-to-digital converter / Capillary electrophoresis / Miniaturization
DOI 10.1002/elps.201400320

 Additional supporting information may be found in the online version of this

article at the publisher’s web-site

1 Introduction
Since the introduction of chromatography in 1903 by Tsvett [1]
and electrophoresis in 1930s by Tiselius [2], separation meth-
ods were upgraded/developed into sophisticated analytical
tools for complex mixtures analysis [3]. Separation systems
are coupled to a detector that detects migrating analytes
through the detection window. Such systems are commonly
purchased as benchtop devices. Numerous suppliers can be
found providing different bench-top size analytical systems
worldwide. Contrary, field-portable, stand-alone, miniatur-
ized separation/detection systems, or modules are described
more often in scientific literature [4]. Demanding industrial
guidelines [5], ambitiously growing scientific community [6]
require automated, miniaturized, wireless, smart systems
for chemical analysis, environmental monitoring, process
control etc.
Numerous scientific articles describe miniaturized
analytical tools and detectors integrated into separation
Correspondence: Professor Audrius S. Maruška, Faculty of Natu-
ral Sciences, Department of Biology, Vytautas Magnus University,
Vileikos 8–206, LT44404 Kaunas, Lithuania
E-mail: a.maruska@gmf.vdu.lt
Fax: +37-037-327908
Abbreviations: CDC, capacitance-to-digital converter; GND,
ground; IC, integrated circuit; LDO, low dropout; PCB, printed
circuit board
systems [7, 8]. Not only separation devices require modern
detection systems, but miniaturized mass analyzers [9], fluo-
rescence detectors [10], spectrophotometers [11], impedance
analyzers [12], capacitive sensors [13], gas sensors [14],
Fourier transform infrared spectrometers [15] as well. The
wireless data transmission implementation into modern
detectors is presented in the scientific literature [16].
Integration of modern detection systems or coupling
them with separation systems is a challenge. Special elec-
trode optimization [17], additional coupling interface [18] etc.
are needed. Frequently, after coupling the device or system
has bigger dimensions [17], or such systems need additional
units for operation (laboratory power supply, alternating
current source, connectors etc.) [19].
Single chip techniques provide numerous advantages:
low noise, low power consumption, small-size of the de-
vice [20]. Therefore capacitive sensor AD7745 integrated cir-
cuit (IC) was selected as a candidate designing the single chip
detector [21]. Manufacturers state, that this IC can be powered
from a low voltage source of 3.3 V and consumes 700 ␮A cur-
rent. Such characteristics provide a possibility for low power
consumption applications.
Often designers/engineers face additional problems,
when designing miniaturized systems. Printed circuit board
traces routing can be a challenge due to cross-talk and other
parasitic effects [22].
Capacitance-to-digital (CDC) technology allows measure-
ment of capacitance and conversion of it to digital code [23].
There are several publications with this technology applied


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2 T. Drevinskas et al.
for liquid holdup measurement [24], detection of volatile com-
pounds [14], structural health monitoring [16] etc. This tech-
nology has already been applied in separation science [25,26].
In our previous work the single chip (AD7745) was used
as whole detector. The device operation required laboratory
power supply, additional electronics prototyping board, and
USB cable for data transmission [27]. Only one single-ended
electrode configuration was investigated in the former set-
up. Differential signal acquisition concept can be applied for
detection. Such technique provides higher resistance to ex-
ternal noise [28]. In addition, Wheatstone bridge is a widely
used measurement technique in the modern devices (digi-
tal scales, pressure sensors etc.). This measurement imple-
ments four wires and four resistances (impedances). Usually
three resistances are equal and known and one is used as a
sensor [29]. This method allows a simple proportional calcu-
lation of unknown resistive value of the sensor. Inequalities
due to the presence of different length wires are neglected,
therefore only the change of sensor resistivity is recorded
[29, 30].
The aim of this work was to design a stand-alone,
battery-powered, wireless, capacitance-to-digital single chip
detector for capillary electrophoresis and the improvement
of detection parameters using different signal acquisition
configurations.
2 Materials and methods
2.1 Chemicals
L-Histidine (L-His) (99.0%), monohydrate MES (98.0%) were
purchased from Alfa Aesar (Germany), sodium hydrox-
ide (NaOH) (99.0%) was purchased from Reachem (Slo-
vakia), potassium dihydro phosphate (NaH2 PO4 ) (99.3%) was
from Fisher Scientific (USA). 2-Amino-2-(hydroxymethyl)-
1,3-propanediol (TRIS) (99.8%) was purchased from Merck
(Germany), sodium chloride (NaCl) was purchased from
Lachema (Czech Republic), acetic acid (99.8%) was purchased
from Lach-ner (Czech Republic). Histamine (⬎97.0%) and
melittin (91.8%) were from Sigma-Aldrich (Germany). Bee
venom was kindly provided by the local beekeeper. Bidistilled
water (produced in a laboratory) was used for preparation of
the solutions.
2.2 Instrumentation
AD7745 chip was purchased from Analog Devices (USA),
LT1761 3.3 V low dropout (LDO) voltage regulator was pur-
chased from Linear Technology (USA). Other electronic parts
and wires were purchased from the local store, APC220 wire-
less data transmission module was from DFROBOT (China).
Sparkfun pro micro prototype development board was pur-
chased from SparkFun (USA). Li-ion (3.7 V, 1800 mAh) cell
phone battery from Kruger & Matz (Poland) was used to
power the device. Analyses were performed using HP 3D CE

C 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2014, 00, 1–6
capillary electrophoresis system (Agilent Technologies, Wald-
bron, Germany). The data acquisition and peak integration
software was programmed in house using open-source soft-
ware Processing 2 (USA). Other calculations were performed
using Microsoft Excel (USA).
2.3 Sample preparation and electrophoretic
separation
Stock solutions of analytes (NaCl, KH2 PO4 , TRIS, histamine,
melittin, and bee venom) and 20 mM MES/L-His buffer/BGE,
pH 6.1 were prepared by dissolving adequate mass of sub-
stance in bidistilled water and frozen at −20°C to pre-
vent microbial growth. BGE of 1 M acetic acid (pH 2.4)
was prepared by diluting adequate volume of acetic acid with
bidistilled water. Before use, the stock solutions were de-
frosted and diluted to required concentration using bidistilled
water. Buffer solution was degassed in vacuum for 20 s before
analysis.
Electrophoretic separations were performed using 50 ␮m
id, 365 ␮m od fused silica capillary. Analysis temperature was
set at 25°C. A 14 and 17 kV voltages were used for detection
evaluation and real-sample analysis correspondingly. Sam-
ples were injected hydrodynamically at 50 mbar for 5 s. Sepa-
ration in MES/L-His buffer was performed in less than 5 min
and separation in 1 M acetic acid was performed in 12 min.
For evaluation of detection parameters 20 mM MES/L-His
buffer was selected as BGE. Metal cations were separated
using TRIS buffer [31]. Capillary was flushed for 2 min with
0.1 M NaOH, bidistilled water, and BGE prior to each analysis.
Each analysis was repeated three times. LOQ were calculated
in accordance with valid pharmaceutical analytical method
validation guidelines [32].
3 Results and discussion
3.1 Design of battery powered detector
Battery holder, holder clips, and detection cell case were
3D-printed using a Printrbot 3D printer (Printrbot, USA).
Microcontroller development board and wireless data trans-
mission module was glued on top of the battery holder us-
ing hot glue. AD7745 chip was soldered on 2-sided printed
circuit board (PCB) that was carefully routed in accordance
with technical documentation [23] and mixed signal lay-
out guidelines [33]. The AD7745 contains digital and ana-
log signal conversion parts, therefore IC placement affects
the designed device performance. The device was designed
of several modules (prototyping board, wireless transmis-
sion module, and AD7745 capacitance detector), therefore
solid/unsplit grounding cannot be applied [33].
In our previous applications the single chip detector
was powered from laboratory power supply [27]. All ground
(GND) wires were connected to the single point of power
plug grounded wire. The battery powered, stand-alone devices
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Electrophoresis 2014, 00, 1–6 Microfluidics and Miniaturization 3

Figure 1. Schematic diagram of the

detector. (A) Single-ended configu-
ration. (B) Differential configuration.
(C) Wheatstone bridge configuration.
Drawn using Cadsoft Eagle (USA).

Figure 2. Photographs of different con-

figuration detectors. (A) Single-ended
configuration, (B) differential configura-
tion, (C) shielded device with copper foil.

cannot be connected to a common grounded wire, therefore of wire/trace inequality and different parasitic values for each
other solutions of this problem should be made. input pin of AD7745. Software code was adjusted so that the
High resolution analog-to-digital converters require a capacitive offset calibration register [22] would have 15 fF
very low noise power supply [34]. Consequently, low noise, offset at start (Supporting Information Vid. 1).
micro-power, LDO voltage regulator LT1761 was used for on-
board voltage stabilization. Stabilized voltage of 3.3 V pow-
3.2 Performance evaluation
ered the AD7745 chip. Decoupling capacitors were selected
as suggested in the datasheet [35]. Power supply of AD7745
First of all the previously reported version of single-ended
was bypassed using 100 nF ceramic, 1 and 10 ␮F tantalum
single chip detector [27] was reproduced (Fig. 2A). Several
capacitors (Fig. 1).
changes were implemented, namely: (i) PCB was etched with
Electrode geometry of the device was similar to geom-
GND layer under all electrode area; (ii) AD7745 chip and
etry used in our previous work [27]. Electrodes were made
passive electronic parts were completely shielded. Unshielded
from stainless steel sleeves (syringe needles) (0.4 mm id,
detector with single-ended configuration resulted in higher
0.6 mm od, 19 mm length) and soldered onto the PCB leav-
noise floor than unshielded detector with differential signal
ing a gap of 0.2 mm between actuator and sensor electrodes
acquisition configuration (Fig. 2B). Resolution parameters
(Supporting Information Fig. 1). Shielded wires were used
were calculated (Table 1).
for power supply from the battery and for digital data con-
nections. Data were collected at 9 Hz rate via inter-integrated
circuit interface and instantly sent to the computer via a wire- Table 1. Comparison of performance of different configuration
less system. The total mass of the designed detector was 82.5 g detectors
(Supporting Information Fig. 2). No. Shield Effective resolution Wheatstone bridge
Three different electrode configurations were tested: (i) (peak-to-peak bits) (Differential)
one electrode couple in single-ended mode (Fig. 1A); (ii) two
electrode couples in differential mode (Fig. 1B); (iii) four Single-ended Differential
electrode couples—Wheatstone bridge (Fig. 1C) (differential
1 Shielded 16.6 17.3 16.7
mode). In differential operation mode offset calibration was
2 Unshielded 13.4 16.7 16.3
performed. This procedure was necessary for compensation


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4 T. Drevinskas et al.
Table 2. Detector performance regarding capillary wash
procedure
Effective resolution (peak-to-peak bits)
No. Flushing solution Active electrode Active and reference
couple washed electrode couples
washed
1 Bidistilled water 17.3 17.3
2 MES/L-His buffer 17.3 17.3
3 0.1 M NaOH 12 17
The results obtained show, that single-ended electrode
configuration is highly susceptible to noise, therefore the de-
vice must be completely shielded (Fig. 2C). The unshielded
device differential electrode configuration offered similar per-
formance to the shielded device.
During detector performance evaluation it was noticed,
that the noise floor of a detector differs when different so-
lutions are flushed through the capillary. The sensor elec-
trode connected to Cin(±) and the high conductivity liquid
(0.1 M NaOH) inside the capillary form a capacitively cou-
pled antenna which degrades signal-to-noise ratio/effective
resolution. However, undesirable background noise can be
reduced by injecting the same liquid into reference electrode
couple. Consequently, two antennas are formed providing
same pattern of noise into Cin+ and Cin– of AD7745 chip
inputs resulting in subtracted noise and higher performance
(Table 2). Similar tendencies showing reduced performance
of capacitively coupled contactless conductivity detector (C4 D)
detectors and higher background noise are presented by other
researchers [36]. The latter issue is the main cause of degraded
performance for single-ended C4 D detectors when high con-
ductivity buffer is used for separation. In another work [30]
slight degradation of effective resolution from 16.8 bits (low
conductivity BGE) to 16.0 bits (high conductivity BGE) bits
was reported.
Another valuable feature of the detector was noticed dur-
ing the capillary cassette temperature stabilization period.
Low baseline shifting was observed when the temperature of
capillary cassette is increased or decreased. The effect of tem-
perature fluctuations was evaluated and compared between
differential and single-ended electrode configurations. Dur-

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Electrophoresis 2014, 00, 1–6
Table 3. Recorded values of capacitance for different electrode
configurations and different flush solutions
No. Solution Configuration Value at Value at Difference
15°C (fF) 30°C (fF) (fF)
1 Bidistilled water Single-ended 145.2 155.7 10.5
2 Bidistilled water Differential 19.2 20.0 0.8
3 20 mM MES/L-His Single-ended 287.1 315.5 28.4
buffer
4 20 mM MES/L-His Differential 9.6 13.5 3.9
buffer
ing temperature fluctuations test the capillaries inside mea-
surement electrode couple (for single-ended configuration)
and measurement/reference electrode couple (for differen-
tial electrode configuration) were injected with the test liquid
(water and 20 mM MES/L-His buffer). Temperature shifts
from 15 to 30°C and vice versa were set. During temperature
change period in capillary cassette the values of capacitance
were recorded. Lower baseline change during the tempera-
ture shifting was observed for differential electrode configura-
tion than for single-ended electrode configuration (Table 3).
The differential signal acquisition mode showed increased
performance during temperature fluctuations.
One of the main problems of contactless conductivity de-
tection (or similar complex impedance measurement tech-
niques) is caused by fluctuating temperature [36, 37]. In
this case, temperature fluctuations affect both: active and
reference electrode couples, therefore, undesirable baseline
shifting is negligible or reduced (Fig. 3).
3.3 Separation and detection evaluation
In our previous work [27] it was shown that CDC detector
is capable of detecting of inorganic and organic cations and
anions in high and low conductivity buffers. Detection capa-
bilities in different BGE solutions were investigated. In this
paper, a low conductivity buffer was tested using model ana-
lytes in order to evaluate performance of the designed device.
Separation of three cationic analytes was performed in 20 mM
MES/L-His pH 6.1 buffer solution. Baseline separation was
achieved in less than 5 min (Fig. 4).
Figure 3. Temperature fluctuations
influence on CDC detector reading.
Temperature shifted down from 30°C
to 15°C and vice versa. Gray line—
single-ended electrode configuration
reading, black line—differential elec-
trode configuration reading. Condi-
tions: 50 ␮m id, 365 ␮m od fused silica
capillary (A) 20 mM MES/L-His buffer
flushed, (B) bidistilled water flushed.
Detection: 3.3 V, 32 kHz.
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Electrophoresis 2014, 00, 1–6 Microfluidics and Miniaturization 5

can be stated, that the measurement is performed only in the

liquid present inside the capillary.
The detector was designed manually (all electronic parts
and electrodes were soldered by hand). The use of sophis-
ticated pick-and-place machines for electrode soldering is
expected to improve the symmetry that should increase
the performance of the detector. Such techniques are ap-
plied in the difference or instrumentation amplifiers, when
symmetry plays an important role on device performance
[28].

3.4 Real-sample separation

Figure 4. Separation of 1–K+ (44 ␮M), 2–Na+ (52 ␮M) ions and
The results obtained by other authors show that bee venom
3–TRIS (28 ␮M). Conditions: 50 ␮m id fused silica capillary,
effective length (Leff )–37.5 cm, total length (Ltot )–50.0 cm, 14 kV can be successfully separated in acidic BGE [38, 39]. In the
voltage potential, 25°C separation temperature, injection at present work, separation of bee venom was performed in the
50 mbar * 5 s. BGE–20 mM MES/L-His buffer, pH 6.1. Detection: BGE of 1 M acetic acid, pH 2.4. Conditions were not optimized
3.3 V, 32 kHz. for separation, since the intention was only to provide data
showing, that separation and detection of complex mixture is
Table 4. Calculated LOQ for different electrode configuration
possible using the detection setup of designed configuration.
detectors
Separation was performed in 12 min (Fig. 5). Analytes were
No. Range (␮M) LOQ (␮M) detected using a CDC detector with differential electrode con-
figuration. The reference electrode couple contained empty
Single-ended Differential Wheatstone fused silica capillary (50 ␮m id, 365 ␮m od). The results ob-
electrodes bridge
tained showed excellent linearity (R2 ⬎ 0.99) of the histamine
K+ 11–27500 0.54 0.31 0.56 calibration curve. LOQ for histamine was 0.4 ␮M. Perfect
Na+ 14–3500 0.49 0.25 0.45 migration time repeatability (RSD ⬍ 1.0%) and good peak
TRIS 7–1875 0.83 0.55 0.83 area repeatability (RSD ⬍ 3.6%) were achieved. Histamine
and melittin peaks were identified in the electropherogram
of separated bee venom using external standard method.
Obtained results showed excellent linearity (R2 ⬎ 0.99) of Determined concentration of histamine in the sample was
the calibration curves for the selected analytes and electrode 40 ␮M and histamine part in bee venom was found to be
configurations tested. High migration time repeatability 1.0%.
(RSD ⬍ 1.2%) and good peak area repeatability (RSD ⬍ 2.2%)
were achieved. LOQs were calculated for all the analytes and
different electrode configurations used. LOQ for K+ were
0.54 using single-ended setup and 0.31 ␮M using differen-
tial electrode setup (Table 4). In the previous work, LOD
(LOD is assumed three times lower than LOQ) of K+ was
1.6 ␮M using the same conditions, which shows an improve-
ment of more than eight times for similar electrode con-
figurations. The improvement of sensitivity is caused by an
implementation of on-board voltage regulator. The total im-
provement of sensitivity of upgraded detector was more than
15 times.
Better performance was expected from Wheatstone
bridge electrode configuration, however the results were very
similar to that obtained using single-ended configuration.
This can be explained by the fact that in Wheatstone bridge
electrode configuration the detection cell was separated from
other electronic parts. Although shielded wires were used Figure 5. Separation of bee venom sample (0.4 mg/mL) dissolved
for signal transmittance, separated modules usually decrease in water. Conditions: 50 ␮m id, 365 ␮m od fused silica capillary,
Leff —37.5 cm, Ltot —50.0 cm, 17 kV voltage potential, 25°C sepa-
performance of the device [33]. ration temperature, injection at 50 mbar * 5 s. BGE—1 M acetic
What is most important, the differential electrode config- acid, pH 2.4. Detection: 3.3 V, 32 kHz. Peaks: 1–40 ␮M histamine,
uration provides a subtraction of parasitic effects, therefore it 2–melittin.


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6 T. Drevinskas et al.
4 Concluding remarks
Stand-alone battery-powered capacitance-to-digital detector
for capillary electrophoresis, capable of transmitting real-time
data wirelessly, was designed. Different signal acquisition
electrode configurations were evaluated, showing approxi-
mately 15 times higher LOQs comparing it to the previous
work. The effect of external noise and thermal fluctuations
on detector performance was reduced. Developed system was
applied for the real-sample analysis.
This research was financed by the Research Fund of Vytautas
Magnus University, grant No. P-BF-13-07. Instrumental sup-
port from Agilet Technologies (donation of HP 3D CE capillary
electrophoresis system) is highly acknowledged.
The authors have declared no conflicts of interest.
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