Alexys Electrochemical System Solutions

Transcription

Edition 2010-2011
EC
ECD
HPLC/EC
Alexys™
electrochemistry and liquid chromatography
PRODUCTs & Parts CATALOGUE
LC-ECD Systems
Detectors
Autosamplers
Pumps
Organizers and racks
Flow cells
Columns, kits and valves
Software
®
Documentation
Company profile
C o m pa n y P r o f i l e
®
For over 20 years Antec has been the world’s leading
supplier of analyzers using High Performance Liquid
Chromatography (HPLC) with Electrochemical Detection
(ECD). With the introduction of the ROXY™ line of products
for on-line Electrochemistry (EC)/MS, Antec is bringing
electrochemistry back to the attention of the chemist,
assuring success with EC/MS applications and the mimicking of nature’s Redox reaction.
Antec is headquartered just outside Amsterdam,
in Zoeterwoude, The Netherlands. Antec offers its products
and services through a worldwide distributor network.
Antec’s subsidiary in Palm Bay, Florida provides products
and services directly to the North American market.
In the early years, Antec’s product offering was primarily
comprised of electrochemical detectors. That changed with
the release of the ALEXYS™ analyzers, a fully featured
HPLC/ECD product line, and the introduction of the
ROXY™ product line for electrochemistry upfront MS.
For Antec, focus on application support and product quality
has always been our highest priority. We have strong
collaborations with our customers developing dedicated
Proven EC-systems to guarantee your performance
and customized application solutions.
Sharing knowledge of the systems and application methods,
including on-site training, has become a valuable service in
supporting our customers. Antec’s commitment to providing
‘guaranteed applications’ means that upon installation
Antec invests time and efforts to have customers’
applications fluidly running ensuring a head start without
losing time over details.
®
2
Index
Index
ALEXYS™ Analyzers products & parts catalogue
Company profile ........................ 2
Index ............................................................... 3
ALEXYS™ Analyzers .......................................... 4
Configuring ALEXYS™ Analyzers .......................................
ALEXYS™ Analyzers ready to prove performance .............
ALEXYS™ Analyzers with fluidly running applications .........
ALEXYS™ Analyzers line-up ................................................................
4-5
6-7
8-11
12-13
Detectors ............................................................................................ 14
DECADE II ............................................................................................................. 14-15
DECADE II SDC ...................................................................................................... 16-17
Autosamplers ............................................................................................. 18
Autosampler AS 100 .................................................................................................. 18-19
Autosampler AS 110 .................................................................................................... 20-21
Pumps ................................................................................................................... 22
Pump LC 110 ..................................................................................................................... 22-23
Organizers and racks .................................................................................... 24
Organizer and racks ........................................................................................................ 24-25
Flow cells .............................................................................................................. 26
Columns, kits and valves ....................................................................... 30
Columns, kits and valves ........................................................................................
30-31
Software ....................................................................................................... 32
Clarity™ .................................................................................................................... 32-33
Dialogue software™ ......................................................................................... 34-35
Documentation ............................................................................ 36-37
Application and technical notes ...................................................... 36
Support ................................................................................................. 37
Distributors ................................................................... 38-39
Proven EC-systems to guarantee your performance
VT-03 electrochemical flow cell ................................................................................. 26-27
FLEXCELL™ ........................................................................................................................ 28-29
Worldwide representation .......................................... 38-39
®
3
ALEXYS™ Analyzers
Validated application
All relevant HPLC and detection parameters such as mobile
phase composition, working potential etc. have been established and optimized for you. The robustness, linearity
and repeatability of the method has been determined for
you by our experts at Antec. The data acquisition software
contains pre-configured system files and acquisition parameters to run any validated application. Application notes are
published on the internet. The system solution is ready for
Proven analyzers to guarantee your performance
you to use straight out of the box.
®
4
ALEXYS™ Analyzers
Configuring ALEXYS™ Analyzers
Proven to guarantee performance
All parts of a system solution have been carefully selected
to meet the requirements for best sensitivity in HPLC with
EC detection. Additional components such as switching
valves or mixing devices are added if necessary.
System configuration
®
5
ALEXYS™ Analyzers
Due to its incredible sensitivity, EC detection has very
high demands from an LC system.
To fully exploit LC-ECD, Antec Leyden has
created a system that meets these
demands.
This dedicated system is
comprised of the DECADE
II, autosampler, pump and
data acquisition software.
An organizer rack with
a high-efficiency integrated degasser, up
to 2 pulse dampeners and a mains
power distributor
for all devices
are also part of
the system.
The ALEXYS™ is
easily upgradeable to allow
micro-bore and
capillary LC-ECD.
The fully featured
data acquisition
software contains
all the tools for automated sampling, data
acquisition, calibration,
processing and reporting.
®
6
ALEXYS™ Analyzers
ALEXYS™ Analyzers
Ready to prove performance
Integrated system solutions have been developed
for the analysis of neurotransmitters such as
noradrenaline, dopamine, serotonin and
metabolites in microdialysates.
Typically, picomolar concentration
detection limits have been
obtained in only a few
microliters of sample.
Parallel detection
schemes using
multiple flow cells
have been
developed for
time, sample and
experimental
cost savings!
The ALEXYS™
can be used
for routine
analyses in
clinical, food &
ronmental and
pharmaceutical
laboratories.
beverage, envi-
®
7
ALEXYS™ Analyzers
Neuroscience
ALEXYS™ Analyzers
with fluidly running applications
Neurotransmitter analyses in microdialysates & brain tissue
For the analysis of noradrenaline, dopamine, serotonin
and acid metabolites two assays are presented in the
application note. One method for dopamine and
serotonin (I), and another method for noradrenaline,
dopamine and metabolites (II). The effect of modifier
content, ion-pair reagent concentration, mobile phase
ALEXYS Monoamines Analyzer™
• All monoamines & metabolites
•One injection, two chromatograms, fully flexible
•Small samples < 5 µL
pH, and electrode potential on the analysis of these
substances are presented. Detection limits are
ALEXYS GABA and Glutamate Analyzer™
typically in the range of 0.2fmol on column.
• Automated pre-column derivatization
NEUROSCIENCE
A P P L I C AT I O N N OT E
213_017 #03
•One injection, two chromatograms,
column switching
Page : 1/6
SEROTONIN, NORADRENALINE, DOPAMINE
AND METABOLITES
IN MICRODIALYSATE
213_017 #03
Page : 3/6
nA
5-HIAA
0.04
Tabel 2
Conditions for DA and 5-HT analysis, without ion-pair reagent
ALEXYS® 100 LC EC system
HPLC
Oven temperature
35 °C (column and detection)
Flow rate
200 µL/min
Flow cell
VT-03 GC 2 mm / ISAAC
ADF™
0.02 Hz
Range
1 nA/V
Using a 15 cm column will further resolve the DA peak, but will
also result in a longer retention time for 5-HT (Fig. 7).
nA
0.02
A
capacity factor
Capacity factor K’
Glutathione and other thiols
HVA
1.0
0.00
0.9
1
2
3
4
5
min
Fig. 5. Chromatogram of 5 µL injection of 1 nM DA and 5-HT standards in
0.01 M PCA. Conditions: Tabel 1.
0.8
0.7
Tabel 1
Conditions for DA and 5-HT analysis, without ion-pair reagent
0
HPLC
2
4
6
8
10
12
14
16
min
Oven temperature
Flow rate
200 µL/min
Flow cell
ADF™
0.02 Hz
Range
1 nA/V
Fig. 7. Analysis of 10 nmol/L NA, A, DHBA, DA, 5-HT standards. Conditions: Tabel 2, but using a 15 cm column and 5% methanol applied.
II. Noradrenaline, dopamine and metabolites
If the selectivity of the method is not sufficient, another set of
HPLC conditions can be used resulting in more retention (Tabel
2). Ion pairing reagent and methanol must be added to optimise
the separation. Under these conditions capacity factors changed
from 1.3 and 8 to 2.6 and 16 for DA and 5-HT, respectively.
For the analysis of noradrenaline, dopamine, HVA, 5-HIAA and
DOPAC a 15 cm column was used. Especially for noradrenaline,
which elutes close to the front peak it is important to have enough
selectivity. Also it must be taken into account that peaks of the
acid metabolites are usually much higher than NA and DA peaks
(see figure 9).
Tabel 3
10
100.0
DOPAC
NA
Dopamine
0
0
0.1
0.2
Noradrenaline
Conditions for the analysis of noradrenaline, dopamine, HVA, 5-HIAA
and DOPAC.
DA
DA
5HT
HVA
75.0
0.3
HPLC
5-HT
1 pA
SOS (g/L)
50.0
Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - [email protected] - www.myantec.com
Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076 - USA - [email protected] - www.myantec.com
Fig. 2. Capacity factors at different ion pair concentrations of sodium octyl25.0
sulfate (SOS).
Although in ion chromatography the concentration of the counter
+
4
ion (Na ) will affect the retention [ ], it is not commonly used as
parameter for optimisation in ion pair chromatography of biogenic
amines.
0.0
0
0
100
200
300
400
2
4
6
8
ALEXYS Acetylcholine Analyzer™
5-HT
8-OH-DPAT
M OP EG
9
HVA
NA
3-nitro-L-Tyrosine
10
DA
10
below the nanomolar and
sometimes even below pico-5
3,4-DHBA
Choline (Ch)
Acetylcholine (ACh)
Markers for oxidative stress
concentrations are often
DA a Limit of Detection (LOD) of 0.2 fmol (0.1 nmol/L) was found.
For 5-HT the LOD was 0.4 fmol (0.2 nmol/L) respectively.
I. Dopamine and serotonin
For the analysis of dopamine and serotonin a simple mobile phase
has been selected without ion-pair reagent or methanol (Tabel 1).
To avoid interfering peaks from acid metabolites a pH of 6 has
been chosen. No ion pairing reagent was used which further
discriminates peaks of amine compounds. Both substances of
interest could be separated easily with enough retention. Column
length is 5 cm for short analysis times. The total run time is less
than 5 min.
DA
Glutamate (Glu)
5-HIAA
The method was optimised for analysis of noradrenaline (NA), 8
5-HIA A
DOP A C
dopamine (DA), serotonin (5-HT) and the metabolites 57
hydroxyindole acetic acid (5-HIAA), 3,4-dihydroxyphenylacetic 6
acid (DOPAC), homovanillic acid (HVA).
5
In particular the combination of the strongly retained 5-HT and
4
noradrenaline, which elutes closely to the front peak, makes
3
quantitative analysis in a reasonable timeframe almost impossible.
2
molar concentration range.
Therefore, samples were split and analysed using two different
1
methods.
One
method
for
dopamine
and
serotonin
(I),
and
The sample volume is usually
0
4
5
6
-1
0
1 method
2
4
another
for3noradrenaline,
dopamine and metabolites
pH of m obile phase
limited by the dialysis flow
Methanol concentration (%)
(II).
Not only the modifier content and detection potential but also
rate (0.5 – 2 μL/min) and the
concentration
ion-pair
reagent
pH are
crucial parameters
Fig. 1. Capacity the
factors
of all substances
are affected
by and
methanol
percentFig. 3. Capacity factors of acid metabolites strongly decrease at high pH.
temporal resolution required.
age.
that were investigated for optimisation of selectivity and detection
limits.
Retention of catecholamines is not affected by pH in the range 3Available sample volume is
Careful adjustment of these parameters is required, as it will
be
6, however
above pH 6 also the retention times of amines intypically between 1 – 15Ion-pair
μL. reagent
impossible to obtain picomolar sensitivity in dialysis samples
crease.
An ion pairingwithout
reagent,prior
suchoptimisation
as hexyl-, octylor dodecylsulfate is
Specific requirements for
of selectivity.
often added to the mobile phase to increase retention of amines.
neurotransmitter analysis
are
Detection potential
The apolar carbon chain will adsorb strongly to a reversed phase.
Modifier
Optimisation of detection potential is one of the first steps in
investigated and relevant
The sulfonic acid group is forming a pseudo-stationary cation
The percentage modifier will typically affect retention of allmethod
sampledevelopment (Fig. 4). A number of excellent papers are
exchange phase. Below pH 7 the amine of the catecholamines will
assay parameters for HPLCavailable reporting voltammetric behaviour of relevant biogenic
components. Increasing the percentage modifier will decrease
be fully
and retained by complex formation with the
ECD method development
areprotonated
and metabolites [1-2-3]. Nagao and Tanimura [1] classiretention times (Fig.1). Acetonitrile [ 1 ] and methanol [ 2 - 3 ] amines
are often
sulfonic acid (Fig. 2). The neutral and acid metabolites however
fied the biogenic amines in four groups depending on their elecused, sometimes in combination. We have a preference for
presented.
are not affected [3].
trochemical
methanol as in our experience certain qualities of acetonitrile
may behaviour, using a mobile phase at pH 3.6 and a salt
bridge AgAgCl reference electrode. These four groups are cateresult in electrode contamination. The use of tetrahydrofuran
40
chol compounds such as the catecholamines, DOPAC and DOPA
(THF) is not5-HT
recommended because it will lead to an unacceptable
(E½ = 380-500 mV), indoles such as 5-HT and 5-HIAA (E½ = 480high background current.
520 mV), Vanillic compounds such as VMA, HVA and MHPG (E½
= 640-680 mV) and monohydroxyphenols such as tryptophan and
30
tyrosine (E½ = 870 mV). It should b • e noted that the given values
are dependent on pH (about 60 mV per pH unit), mobile phase
composition and differences in glassy carbon working electrode
DA
20
material.
HVA
Although absolute values are different (smaller) at pH 6 we found
the expected relative differences in E½ for DA, 5-HT and HVA.
detection. Neurotransmitter
homovanillic acid (HVA)
Choline and Acetylcholine
Method
sis device and analysed by
HPLC with electrochemical 15
capacity factor
5-hydroxyindole acetic acid (5-HIAA)
3,4-dihydroxyphenylacetic acid (DOPAC)
4-aminobutyrate (GABA)
DOPAC
20
sampled trough a microdialy-
Serotonin
decrease at high pH (Fig. 3). In fact, at pH 6 these metabolites
elute in the un-retained front peak.
NA
brain of conscious animals is
Dopamine
OPA derivatized amines and amino acids
GABA and Glutamate
25
DA
rotransmission in living brain.
Cerebrospinal fluid of the
Page : 2/6
5-HT
• HPLC with electrochemical detection
• Microdialysis of neurotransmitter in vivo
• Neurotransmission in living brain
• Cerebrospinal fluid of the brain of conscious animals
pH
• Investigated and relevant assay parameters
The retention times of acid metabolites HVA, 5-HIAA and DOPAC
Microdialysis of neurotransinvaluable tool to study neu-
Noradrenalin
SEROTONIN, NORADRENALINE, DOPAMINE AND METABOLITES
IN MICRODIALYSATE
Page : 2/6
INTRODUCTION
mitters in vivo has become an
Monoamines and the metabolites
213_017 #03
SEROTONIN, NORADRENALINE, DOPAMINE AND METABOLITES
IN MICRODIALYSATE
v
THE SMARTEST LC-EC APPLICATIONS FOR
NEUROSCIENCE ANALYSIS
EVER MASTERMINDED
•Enzyme coated electrode
•Superior reproducibility and sensitivity
Oven temperature
Flow rate
200 µL/min
Flow cell
ADF™
0.02 Hz
Range
1 nA/V
10 m in
500
E (mV)
Fig. 4. Hydrodynamic voltammograms of DA and 5-HT. pH of mobile phase:
6.0, reference electrode ISAAC™ (8 mM KCl).
Fig. 6. Pooled frontal cortex microdialysate (red) and microdialysate spiked
with 2 nM DA and 5-HT (blue). Conditions: Tabel 2.
Detection limits were calculated as the concentration resulting in a
signal that is 3 times the peak-to-peak noise of the baseline. For
On-line in vivo analyses
Conventional sampling of once every 20 minutes is not
sufficient to accurately describe fast neurochemical
responses. To improve time resolution, a 14-port valve
with 3 loops connected in series is used to directly
collect the microdialysate. The content of these loops
is then simultaneously injected on 3 HPLC flow paths
• High throughput
• 3 rats, 3 x 6 - port valve
•Efficient multiple analyses on one system
that run the same analysis, and during analysis the
following three samples are collected. A DECADE II™
Triple Cell Control (TCC) detector acquires all signals.
ALEXYS OMD Time Resolution Analyzer™
• 1 rat, 14 - port valve, 3 loops
•Increased sampling frequency
• 3 µL loops = 3 min samples
ALEXYS OMD Monoamines Analyzer™
• All monoamines & metabolites
• 14 - port valve, 2 loops, no sample loss
•Immediate results
®
8
ALEXYS OMD High Throughput Analyzer™
ALEXYS™ Analyzers
pharmaceutival & Biotech
ALEXYS™ Analyzers with fluidly running applications
ALEXYS Aminoglycosides Analyzer™
• Aminoglycosides
•EP/USP methods
•Bulk and veterinary cocktails
217_018 #00
SPECTINOMYCIN
IN PHARMACEUTICAL PREPARATIONS
217_018 #00
SPECTINOMYCIN
IN PHARMACEUTICAL PREPARATIONS
Page : 2/4
Page : 3/4
Page : 2/4
The analyzers are dedicated and methods are
or USP. Aminoglycosides such as Gentamicin,
Lincomycin in cocktails can be analyzed.
CONCLUSION
The ALEXYS® Aminoglycosides analyzer provides a reliable
In the EP monographs for Spectinomycin [4.5] two system
suitability requirements are specified:
[1] Peak resolution: between impurity E and the principle peak
(Spectinomycin), R > 1.5.
#07the principle peak, n=6
[2] Repeatability: maximum RSD(%)
for
217_010
injections, RSD > 3%.
In Table 1 the criteria of the EP are compared with the typical
results obtained with the ALEXYS Aminoglycosides analyzer.
validated to meet the specifications of the EP and/
Tobramycin, Kanamycin, Spectinomycin and
L
TICA E
T
AC E U
ARM TION NO
A
& PH
ECH APPLIC
B I OT
EP requirements
Parameter
EP criteria
RSD of principal peak
Resolution, peak ‘A’
< 3.0 %
> 1.5
maceutical Preparations. It meets the EP system suitability
requirement for resolution and repeatability.
12
YCIN
1.0%
NEOM UG
2.5 S
BULK DR
Solutions and standards are prepared as described in the EP
method [4,5]. Assay validation was done with special attention
to EP requirements.
TIONS
FOR
APPLICA
SIS
H ANALY
BIOTEC
LE LC-EC
RELIAB
ICAL &
ACEUT
PHARM
LATED
FORMU
EVER
THE MOST
ides
Aminoglycos
Amikacin
te
Sulpha
Framycetin
te
icin Sulpha
Gentam
te
ycin Sulpha
Kanam
ycin
Lincom
cin
Neomy
in
Spectinomyc
Tobram
ycin
Macrolide
tics
antibio
mycin
cin
thromy
in
Clarithromyc
Erythromycin
in
s
Roxithromyc
pharmaceutic
lysis of
Bioana
Artemisinin,
isinin,
Dihydro-artem
ther
Arteme
ide
Etopos
PAT
8-OH-D
87
BNP77
mesna
Azithro
Azaery
Vincris
tine
1/5
solution for the routine analysis of Spectinomycin in Phar-
YCETIN
AM
AND FR
Result
It is evident from table I that the EP requirements for both peak
resolution and repeatability are met by the ALEXYS aminoglycosides
analyzer.
Fig. 2. Overlay (n=6) of 20 μL injections of 80 mg/L
Spectinomycin-HCl in mobile phase (diluted from 800 mg/L
Spectinomycin in water, standing time 68 h).from paper [2]
and based on peak area percentages.
Page :
TE IN
SULPHA
217_018
#00
B I OT
AL
UTIC
M AC E N N OT E
PHAR
IO
E C H & A P P L I C AT
/4
Page : 1
S
ION
EPARAT
OMYCIN
ICAL PR
SPECTIN
MACEUT
IN PHAR
•
TION
ODUC
iotic
INTR
is an antib
Neomycin
of a mixconsisting
s
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oglycoside
the amin
ture of
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describces
Neomycin
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graphs
D.
3
two mono
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LC-PA
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tained
(EP) has
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[ ][ ][
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FOR
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THE MOST
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oglycoside
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PHARM
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EVER FORMU
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application
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ycin B).
vanalyzer
solution
requiremen
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peak (Neom
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in infec
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analy
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It
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note typica demonstrat
lycosides
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available
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in bulk
s, gels,
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impurities
(e.g., cream
Sulphate
. FramyceFramycetin
eye drops
te
ycin
etc.) and
icin Sulpha
Method
as Neom
Gentam
known
oSulphate
tin (also
aminoglyc
Kanamycin
no
Tabel 1
er(part
ate) is an
ions
analyz
Condit
B sulph
Lincomycin
ar to
LC-EC
glycosides
iotic simil
S amino
n
ALEXY Neomycin
side antib
monly
50A)
0.5 mL/mi
com180.00
olumn:
omycin
and
™
Spectin
HPLC
n, post-c
HyREF
name
Neomycin
0.7 mL/mi
WE and
ycin
r the brand
with Au
ll™Tobram
detection
in
tics
and
sold unde
Flexce
Flow rate
tion
rities
antibio
de
separa
. Impu
Macroli
for
32 �C
Flow cell
Soframycin
cetin
e
Azithromycin
and framy
10 µA/V
cin
Temperatur
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thromy
neomycin
0.5 Hz Azaery
s are analy
Range
,
-300 nA omycin
preparation
e HPLC
-800 - Clarithr
ADF
sed phas
Erythromycin
using rever
I-cell
NaOH addicolumn
tric
Roxithromycin
ceutics
with postamperome
of pharma
pulsed
Bioanalysis
tion and
[1-3].
inin,
Artemis
(LC-PAD)
sinin,
detection
des
glycosides
• Amino
ION
ds
methods
DUCT
S metho
ils
EP/UPS
INTRO
• EP/UP
cocktails
aminognary cockta
cin is an
veterinary
and veteri
Bulk and
proSpectinomy
• Bulk
e antibiotic
lycoside-lik
es specStreptomyc
duced by
spectinosolution,
tabilis. In
a ring
undergo
will
ALEXYS®
mycin
g of the
with the
obtained
ed, demand closin
Method
ing
l results
are report
opening
on, result
note typica a C18 column
icin.
ation
functi
on
re of
hemiketal
sis of Gentim
In this applic analyzer based
the analybrium mixtu
Gentamycin performance for
in an equili
ers.
its
ble anom
onstrating
four possi
producwith acid
Hydrolysis
1
basic
in
Table
mine and
r
Conditions
analyze
es actina
inoic
LC-EC
lycosides
actinospect
S Aminog
ALEXY
)
solutions
Impord.
.0050A
mL/min
is forme
(part no.180
HPLC
lumn: 0.5
™
acid (ASA)
, post-co
HyREF
impurities
1 mL/min
WE and
ntation
with Au
on
tant ferme
ycin
Flow rate
Flexcell™
and detecti
rospectinom
separation
35 °C for
Flow cell
are dihyd
omycin
roxyspectin
Temperature
2 μA/V
nce
and dihyd
the prese
0.5 Hz
Range
ause of
[1, 2].Bec
μA
s in SpecADF
about 2
side group
of glyco
ducts,
I-cell
EP
and by-pro
bed in the on
tinomycin
rometric
l attenti
red as descri
ampe
d
specia
prepa
are
with
pulse
rds
done
LC with
been
and standavalidation was
(PAD) has
Solutions
-artemi
[4,5]. Assay.
detection
Dihydro
method
sis
ments
for analy
to EP require
Artemether
applied
in corres
itions are
Etoposide
[3]. Cond
requirewith EP
8-OH-DPAT
7
pondence
Antec Leyden B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands
- [email protected]
- www.myantec.com
YS®
The ALEX
mesna BNP778
is
zer’
ments [4,5].
tine
Vincris
osides analy
‘Aminoglyc
on for
LC soluti
a dedicated
inomysis of Spect
the analy
the EP
matches
cin, which
resoluts for peak
the
requiremen
tability of
aand repea
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this applic
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peak. In
www.m
.com
.com principal
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ALEXYS Azithromycin Analyzer™
•Macrolide Antibiotics
•USP method
• Applicable for Clari-, Roxi-, and Erythromycine
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The USP method for Azithromycin utilizes two flow
cells in series. The first cell is used for screening,
and the second for detection of the analytes.
For this application, the ALEXYS Azithromycin Analyzer™
utilizes a FlexCell in series with a VT-03 cell for detection.
Proven
analyzers
to guarantee
your performance
Proven
analyzers
to guarantee
your performance
USA
NV - Zoeterw MD 21076 r
12 - 2382
Hanove
Drive Industrieweg
B.V. Connelley
Antec Leyden Suite 110 - 7502
Antec (USA)
®
9
ALEXYS™ Analyzers
Alexys
110
clinical
& diagnostics
Standardised
routine analysis
ALEXYS™for
Analyzers
ALEXYS Catecholamines Analyzer ™
• validated concept
• robust, dedicated for routine analysis
• tumor and cardiovascular markers
C L I N I C A L & D I AG N O S T I C S
214_001 #01
Today, LC-ECD has been established as a fast and
Page : 1/4
CLINREP® KIT FOR THE ANALYSIS OF
CATECHOLAMINES
IN URINE
reliable method for the determination of catecholamines
and metabolites in plasma and urine as well for
THE SOUNDEST LC-EC APPLICATIONS FOR
CLINICAL & DIAGNOSTICS ANALYSIS
EVER BUILD
INTRODUCTION
The catecholamines norepinephrine (NA), epinephrine
Serotonin. Complete kits contains all the necessary
• Important part as neurotransmitters
• Tumors of the sympathoadrenal system
(A) and dopamine (DA) are
synthesised in the chromaffin
Catecholamines
cells of the adrenal medulla
Serotonin
and the sympathetic nervous
Metanephrines
system and play an important
VMA
chemicals and (calibration) materials for sample
HVA
part as neurotransmitters and
5-HIAA Homocysteine
in metabolic regulation. Tu-
Glutathione
mors of the sympathoadrenal
(di-)sulfides
system cause an elevation in
Iodide
production and in urinary
Vitamins A, C, D, E, and K
excretion of catecholamines
Q10
preparation and analysis of 250 plasma assays,
Ubiquinols
Today, LC-EC has been established as a fast and reliable method
for the determination of catecholamines and metabolites in plasma and urine [1-5].
The ClinRep® complete kit for catecholamines in urine of Recipe
GmbH (Munich, Germany) is a standardised kit for the sample
preparation and routine analysis of urinary catecholamines. In this
application note the results are reported from analysing urine
samples using a ClinRep® kit with an ALEXYS 110 LC-EC system.
and their metabolites.
Method
The quantitative determina-
One Recipe ClinRep® complete kit contains all the neces-sary
chemicals and (calibration) materials for sample preparation and
analysis of 100 assays, excluding the analytical column. Urine
samples are processed using the following sample preparation
procedure prior to analysis:
tion of urinary catecholamines is a rapid and precise
diagnostic method for the
identification of pheochromo-
excluding the analytical column.
cytoma and other tumor diseases of the nervous system
such as neuroblastoma and
214_001 #01
ganglioneuroma. Approx-
214_001 #01
CLINREP® KIT FOR THE ANALYSIS OF CATECHOLAMINES
IN URINE
imately, half of all pheochroIN URINE
mocytoma patients suffer
from permanent hypertension, in rest episodic hyper-
Page : 2/4
Page : 2/4
tensive crises occurs. In
about 40 % of the latter group
• 3 mL acidified urine sample (10 mL conc. 32% HCl per liter
urine) or urine calibrator is mixed with 10 mL stabilising reagent S and 30 μL internal standard (IS) and subsequently adjusted to a pH 3.0 – 7.0 using 0.5M NaOH.
• The mixture is applied to a ClinRep® sample preparation columns to trap the catecholamines present in the sample.
• The column is subsequently washed with 15 mL HPLC-grade
water to remove interfering compo-nents.
• 6 mL of eluting reagent E is then used to elute the catechola: 3/4
mines from the extraction Page
column.
• The eluate is collected, mixed and 20 μL injected in the LC
system.
plasma cate-cholamine concentrations are not raised in
For quality control of the analytical determination Recipe
ClinChek® urine controls have been used in both the normal
(level I) and the pathological range (level II).
1.8
10
12
14
min
An internal standard method is used to compensate for recovery
losses during the sample preparation step. To every urine sample,
calibrator or control 30 μL of internal standard (IS) solution is
added. The IS response of the samples is compared to that of a
directly injected standard solution (ClinTest® standard) to determine the recovery. The sample response is then interpolated to
100% recovery to establish the real catecholamine concentration
in the urine samples.
Table 1
0.8
0.4
0.2
0
2
4
Component
6
8
Flow cell
Column
10
12
14
Norepinephrine
2.8
1.7
16.0
Dopamine
3.7
102.5
Norepinephrine
Table I. Calculated concentration of urine controls level I and II
(n=6). Concentration range specified by Recipe is given for reference (source: data sheet supplied with controls).
Specified conc (µg/l)
Min
Max
Calculated
RSD
14
2.3
conc (µg/l)
(%)
58.7
0.8
14
21
18.7
0.8
120
180
176.6
0.8
Noradrenaline
125
187
156.9
0.2
Adrenaline
29
43
35.1
1.5
Dopamine
186
278
236.9
0.5
Dopamine
44
LC-EC conditions
Flow rate
1.0 mL/min
Sample
20 µl, extracted with ClinRep® sample preparation columns
ClinRep® catecholamine buffer#
Mobile phase
Temperature*
TD2 SDC 30°C (separation & detection),
E-cell
TAS110 4°C (sample cooling)
Range
500 mV (vs. Ag/AgCl sat’d)
I-cell
10 nA/V
ADF
Ca. 0.2 – 3.0 nA
Analysis time
0.1 Hz
Flow rate
5.1
3.2
225.1
66
The ClinRep® complete kit for catecholamines in urine pro-
3.6
vides a standardised method for the sample preparation of
urine samples and fast & reliable analysis of urinary catecholamines using LC-EC.
nA
1.2
Control level I
Noradrenaline
6.2
Dopamine
CONCLUSION
20.7
115.2
nA
1.0
0.25
0.20
0.15
0.10
0.8
0.4
0.2
0.05
2
3
4
5
6
7
min
1
0.0
2
Table 2
2.5
Epinephrine
Dopamine
1.4
Adrenaline
Epinephrine
The RSD’s for norepinephrine and dopamine were smaller then
4%. For epinephrine, which was present in the sample in a significantly lower concentration, the RSD was slightly higher, 6.2%.
30.6
Sample B
min
Fig. 2. Overlay of 6 chromatograms of 20uL injections of
ClinChek® control level I.
Component
0.6
ClinRep® Analytical column for catecholamines
in urine
Table III. Inter-assay precision (urine sample C,
Catecholamines
n= 4 (samples) x 2 (duplicate injections)
x 2 (days).
crisis.
Conc. (µg/l)
Epinephrine
Control level II
ALEXYS 110 LC-EC system with DECADE II
SDC (p/n 190.0035)
GC type flow cell with Ag/AgCl saltbridge REF
RSD (%)
Sample A
Set-up
HPLC
logically increased values,
even after a hypertensive
Component
RSD (%)
Conc. (µg/l)
Table II. Intra-assay precision of urine sample A and B,
Sample C
n= 5 (samples) x 2 (duplicate injections).
48.7 - USA - [email protected] - www.myantec.com
Antec (USA) - Suite 110Norepinephrine
- 7502 Connelley Drive 3.8
- Hanover MD 21076
1.2
1.0
0.6
The quantification of the catecholamines in the urine samples is
performed by means of a single-point calibration method using a
urine calibrator. The ClinCal® urine calibrator supplied in the
ClinRep® kit is a lyophilised urine sample with a known amount
of catecholamines. The urine calibrator should be processed via
the same sample preparation method as the urine samples. An
example chromatogram of a urine calibrator analysis is shown in
figure 1.
worked-up 4 times and analysed (duplicate injection), this procelevels dure
in the
24-hours urine
was repeated the next day and the relative standard deviathecalculated.
detection of pathoThe intra-assay precision of the method was determined using twoallowstion
Norepineph
8
Internal Std
6
Norepinephrine
4
Epinephrine
2
To determine
the inter-assay precision an urine sample (C) was
mination
of catecholamine
urine samples (A and B). The urine samples were worked-up 5
times and duplicate analysis were performed to determine the
relative standard deviation (RSD, %).
1.6
1.4
0.0
0
crises. Nevertheless, deter-
Dopamine
Epinephrine
0.5
Dopamine
nA
1.0
the interval
between
cholamines
in thetwo
concentration range from 1 – 1000 μg/L [6].
Urine samples were collected from an apparently healthy volunteer. and analysed multiple times to determine the recoveries,
LOD, intra- and inter-assay precision of the method.
Epinephrine
IS
1.5
Fig. 1 Analysis of 20 μL ClinCal® urine calibrator. Concetration of
catecholamines in the calibrator sample: 123 μg/L NA, 29.5 μg/L
A and 227 μg/L Dopamine.
noise. The method is linear for the determination of the cate-
Analysis of urine samples
Internal Std
Norepinephrine
2.0
Analysis of ClinChek® controls
Norepinephrine
Epinephrine
Dopamine
nA
2.5
The control samples are lyophilised urine samples which have to
be processed in the same way as the urine samples. Both Control
I and Control II were analysed and the analyte concentrations
quantified using the ClinCal urine calibrator. For both urine controls level I and II the determined concentrations were within the
concentration ranges specified by Recipe on the urine control data
sheet (see table I).
15 minutes
#) mobile phase was recycled during experiments. *) minimum actual oven & tray
temperature which can be reached is dependent on ambient conditions.
4
6
8
10
12
14
min
Fig. 3. Overlay of 10 chromatograms of 20uL injections of urine
sample B. Top-right: zoom in on NA and A peaks.
The RSD’s for all components were typically smaller then 4%.
Only for low concentrations of epinephrine, near the limit of quantitation, a RSD of 14 % was found.
For all urine samples, controls and calibrator recoveries typically
in the range of 85 – 95% were found, compared to a directly
injected standard. The concentration limit of detection (CLOD) for
the method was approximately 1 μg/L for all catecholamines. The
CLOD here is based on a 20 µL injection and defined as the concentration that gives a signal that is three times the peak-to-peak
ALEXYS Iodide Analyzer ™
•Iodide in Urine
• diagnosis of thyroid dysfunction
•Excellent sensitivity and peak shape with Magic Diamond
212_003 #01
ERROR! REFERENCE SOURCE NOT FOUND.
Page : 3/4
ALEXYS Iodide Analyzer™
C L I N I C A L & D I AG N O S T I C S
212_003 #01
Page : 1/4
IODIDE IN URINE
interesting new electrode material became available
in electrochemistry. Diamond has several
ALEXYS Iodide Analyzer
THE SOUNDEST LC-EC APPLICATIONS FOR
CLINICAL & DIAGNOSTICS ANALYSIS
EVER BUILD
PART NUMBERS AND CONFIGURATIONS
190.0035
ALEXYS 110 LC-EC system with DECADE II SDC
INTRODUCTION
A method for the determination of iodide is developed
115.4150
SenCell GC, salt bridge, 5/60
using electrochemical detec-
RE.2000
ClinRep® complete kit , Catecholamine in urine
tion (ECD) with a silver work-
RE.2030
RE.8021
ClinRep® Analytical column
Catecholamines
for catecholamines in urine
Serotonin
ClinChek® urine control, level I
Metanephrines
VMA
ing electrode (WE). The mechromatography using a
phosphate/citrate buffer at pH
5-HIAA Homocysteine
6.5, followed by amperometric
Glutathione
detection at 0.15 V.
(di-)sulfides
Detection limit of iodide is
Iodide
Ubiquinols
Iodide in Urine
Diagnosis of thyroid dysfunction
Excellent sensitivity and peak shape with
Magic Diamond
thod consists of ion exchange
HVA
Q10
•
•
•
0.2-1 µmol/L (25 - 127 ng/mL)
depending on the column
performance. Reproducibility
of the determination of iodide
is concentration depended. At
50 µM RSD in peak area’s and
heights was better than 2%.
Below 1 µM the RSD values
increased to 10 – 22 %.
Method
The HPLC system consists of a anion exchange column with a
phosphate/citrate mobile phase (Table I). The use of halide ions
such as chloride and bromide must be avoided as they are reactive towards the silver electrode, causing a high background
current and decreased sensitivity.
Di-sodium phosphate and citric acid (both 10 mM) are dissolved in
800 mL water, pH is set to 6.5 with NaOH and water is added to
900 mL. Finally 10% methanol (100 mL) is added.
Determination of analytes in complex matrices such as urine or
plasma is often complicated by sample pre-treatment procedures
to improve the selectivity of a method. Without suitable pretreatment co-eluting peaks make reliable and reproducible analysis impossible.
An exception is the determination of iodide in urine. Due to the
selectivity of the silver working electrode, urine could be injected
directly. After dilution and filtration over a 0.2 µm membrane filter,
urine samples have been injected onto the analytical column and
analysed.
advantages over conventional electrode materials
such as a wide potential window in aqueous
solutions, excellent chemical inertness and
stability. For this application a sensitive and reliable
LC-ECD method is available for the analysis of
iodide based on DC amperometry using the
Magic DiamondTM (MD) electrode.
®
10
With the introduction of conductive diamond an
ALEXYS™ Analyzers
Alexys
FOOD
& 110
BEVERAGE
Standardised
for routine analysis
ALEXYS™ Analyzers
ALEXYS Carbohydrate Analyzer™
• QC of food products
• Food research, phytochemicals
• Detect Contaminants and pollutants in beverages
220_002 #03
FOOD
A P P L I C & B E V E R AG E
AT I O N
N OT E
ANALYSIS
OF
ANION-EXC CARBOHYDRATES
HANGE CHR
PULSED AM
OMATOGRA
PEROMETR
PHY AND
IC DETECT
ION
Page : 1/10
THE FINEST
LC-EC
FOOD & BEVERAG APPLICATIONS
FOR
EVER PROCESS E ANALYSIS
ED
The ALEXYS Carbohydrate Analyzer™ gradient system
Bisphenol
A
Catechins
Flavonoids
and phenols
Phenols
Antioxidants
is suitable for the more demanding analysis of complex
Polypheno
ls
Resveratrol
Epicatechin
Quercetin
other polypheno
carbohydrate mixtures such as oligosaccharides.
ls
Carbohydrates
Iodide
Vitamins A,
Q10
C, D, E, and
K
ubiquinols
220_002 #03
Page : 3/10
[A] The ALEXYS 100 'Carbohydrates I' isocratic system
(p/n 180.0052)
A complete system solution consisting of an ALEXYS 100 LC-EC
system with He-proof solvent bottles, ion-exchange column,
guard column and FLEXCELL.
se
co
cro
cto
Glu
La
Su
lto
Ma
oligosaccharides and other complex carbohydrate
se
se
Su
cr
[B] The ALEXYS 100 'Carbohydrates II' gradient system
(p/n 180.0054)
&
se
co
ct
cto
Fru
•
Prepare the NaOH mobile phase using a 50% w/w carbonate-free NaOH stock solution (commercially available) and thoroughly degassed deionised water (≥ 18
MΩ). Commercially available NaOH pellets are not acceptable for eluent preparation, because they are always covered with a thin adsorbed layer of sodium carbonate.
•
The mobile phase should be stored in plastic containers
instead of glass containers. NaOH is a strong etching
agent and will react with the inner glass wall resulting in
the release of silicates and borates.
•
Most dioxide dissolved in deionised HPLC grade water
(> 18 Mohm-cm) can be removed by degassing the water in an ultrasonic bath for 10 – 15 minutes, and subsequent sparging with Helium 5.0 gas.
•
Subsequently add the appropriate amount of 50% w/w
NaOH solution to obtain the final eluent. Always pipette
the necessary amount of NaOH from the top part of the
50% NaOH stock solution. Any carbon dioxide present
in the solution will precipitate as sodium carbonate to
the bottom of the flask, leaving the top part of the solution virtually carbonate free.
•
High-purity grade sodium acetate should be used for the
preparation of the mobile phase (impurities can cause
large baseline shifts during a gradient run).
Ma
lto
La
Glu
mixtures can be recorded. This can serve as a tool for
Glu
Su
Fru
co
se
cto
se
cro
La se
cto
se
A
Carbohydrates
C
0
Ma
lto
se
Antec Leyden
B.V. - Industriew
B
Antec (USA)
eg 12 - 2382
- Suite 110
NV - Zoeterwo
- 7502 Connelley
ude - The
Drive - Hanover
Netherlands
- info@mya
MD 21076
ntec.com
- USA - info.usa@
myantec.
- www.mya
ntec.com
com - www.mya
ntec.com
10
20
The high-pressure gradient system, has in addition to system
above one additional LC 100 pump for HPLC gradients and the
OR 100 organiser rack 'dual channel' option (extra pulse damper).
before the start of the actual current measurement assures that
the contribution of the electrode charging current caused by the
potential steps is small. A large positive potential E2 is applied for
a period t2 to achieve anodic formation of surface oxides in order
to clean the electrode surface. During t3 the oxidized electrode
surface is reduced back to a reactive Au surface, the so-called
reactivation step. So within every individual pulse cycle, the carbohydrate signal is measured and the working electrode surface
cleaned and reactivated resulting in a reproducible detector response over time. Typical settings used in our laboratory for the
analysis of carbohydrates are given in Tabel 1. In this specific
example the total pulse duration is 1000 ms (1 Hz).
A 30 m M N aO H - 10 m M NaO Ac
B 30 m M N aO H - 2 mM NaO Ac
C 30 m M N aO H - 1 mM NaO Ac
estimating chain length distribution.
Fig. 4. ALEXYS 100 ‘Carbohydrates II’ gradient , including two pumps,
column, flow cell and LC connection kit.
Materials
•
•
30
M inutes
Fig. 1. [A] Retention times of common food carbohydrates as a function of
sodium hydroxide (top) and [B] sodium acetate (bottom) concentration of
the mobile phase.
Sodium acetate is commonly used to decrease the elution time of
higher molecular weight carbohydrates allowing faster analysis.
Acetate is a strong competitor for active binding spots resulting in
less interaction of the carbohydrates with the ion-exchange groups
of the column (see Fig. 1B). Sodium Acetate is also particularly
suitable for gradient elution. Pulsed amperometric detectors are
relatively insensitive to ionic strength changes of a sodium acetate
gradient, as long as the sodium hydroxide concentration remains
constant during the gradient run. High purity grade sodium acetate
should be used for the preparation of the mobile phase (impurities
can cause large baseline shifts during a gradient run).
Pulsed Amperometric Detection (PAD) - DC amperometry is not a
particularly suitable method for the electrochemical detection of
carbohydrates. The response of the working electrode is quickly
attenuated, as a result of strong adsorption of the oxidation products onto the working electrode surface.
Detailed information about waveform optimisation for pulsed
amperometric detection of carbohydrates can be found in several
papers published by LaCourse and Johnson [ 3 , 4 ]. The optimum
oxidation potential (E1) for the carbohydrates can be obtained by
cyclic voltammetry, which gives the current-voltage (I/E) relationship of the analyte. The cleaning potential (E2) should be high
enough and the time duration long enough to remove the carbohydrate oxidation products completely. In alkaline media gold
oxide is already formed at E > + 200 mV (vs. Ag/AgCl). At a higher
potential the formation of a metal oxide layer is accelerated and a
shorter time setting can be chosen. It is essential that the time
duration (t3) and the magnitude of the reactivation potential (E 3) be
such that the metal oxide layer is completely removed. Reductive
dissolution already occurs at E3 < 0 mV, but a more negative
voltage speeds up this process. However a too low potential will
result in the chemical reduction of other compounds such as the
O2 dissolved in the mobile phase for instance, leading to baseline
instabilities. The S/N ratio can be optimised by adjusting t1 and
tsample. In principle the time delay determines the level of the background current. In practise, often a delay time (t delay) of 100 – 400
ms is used, and a sample time of 20 – 200 ms.
•
•
•
•
HPLC grade water, resistivity > 18 MOhm.cm
Sodium Hydroxide 50 wt% solution, pro-analyse,
BOOM, the Netherlands
Sodium Acetate trihydrate, >99%, J.T. Baker, the Netherlands
Helium 5.0, 99.999%, Messer, the Netherlands
Mono- and disaccharides standards, Sigma, USA
Maltodextrin, 96.9%, artificial sweetener, Belgium
Mobile phase preparation
Sodium carbonate is a stronger anion than sodium hydroxide. The
formation of carbonate in the mobile phase is the most common
cause of loss of analyte retention. Carbon dioxide gas present in
2air will dissolved as CO3 ions in the strong alkaline eluent. The
dissolved carbonate ions will increase the ionic strength of the
mobile phase resulting in a shortening of the retention times of the
carbohydrate analytes. Therefore, keeping the mobile phase free
of carbonate is one of the key factors towards reproducible carbohydrate analyses via Anion-Exchange Chromatography.
It is advisable to prepare fresh mobile phase daily. The mobile
phase should be sparged continuously with Helium during the
analysis to obtain reproducible retention times.
ALEXYS Phenol Analyzer ™
• Water and Soil samples
• Ketones in biodiesel exhaust
• Used by CRO labs for routine analysis
E N V I R O N M E N TA L
216_001 #03
Page : 1/4
PHENOLS
IN WATER
Chlorophenols and nitrophenols are used for industrial
and agricultural reasons, thus posing a threat to
contaminating river and drinking water. The MAC
(maximum admissible concentration) in the EEC
countries for phenols in drinking water is 0.5 μg/l.
THE BRIGHTEST LC-EC APPLICATIONS
ENVIRONMENTAL ANALYSIS
EVER PLOTTED OUT
FOR
INTRODUCTION
Chlorophenols and nitrophenols are used in industry and
agriculture for several purposes. In the end they may be
found in river or drinking
Chloro- and nitrophenols
2,4-dinitrophenol (DNP)
water. The MAC (maximum
admissible concentration) in
phenol (P)
4-nitrophenol (4-NP)
2-methyl-4,6-dinitrophenol (MDNP)
2-chlorophenol (2-CP)
2-nitrophenol (2-NP)
2,4-dimethylphenol (DMP)
4-chloro-3-methylphenol (CMP)
2,4-dichlorophenol (DCP)
2,4,6-trichlorophenol (TCP)
pentachlorophenol (PCP)
Ketones
the EEC countries for phenols
in drinking water is 0.5 µg/l. In
the 70’s the US environmental
protection agency (EPA)
made a list of the eleven most
important phenol contaminants as priority pollutants.
The standard EPA method is
based on a concentrating
liquid-liquid extraction followed by derivatisation and
GC analysis with electron
capture detection.
In this application a HPLC
method for the analysis of the
11 EPA phenols in water is
described using electrochemical detection. Detection limits
agriculture..
• Chlorophenols in industry and
• Nitrophe-nols in industry and agriculture
• 11 EPA phenols in water
Method
by constructing I/E relationThe detection potential is optimised
of
poor detection characteristics
ships for 9 phenols. Due to the
DNP, a working potential of 1200
some phenols, especially 2,4
experiments. The background
mV vs. Ag/AgCl is used for further
nA.
current is approximately 200
was only problematic at high
Working electrode contamination
in samples are in the
phenol concentrations. The concentrations
level no significant contaminappb
the
At
lower.
or
range
low ppb
day.
could be measured within one
tion of the working electrode
end of each day is sufficient to
One cleaning procedure at the
conditions.
maintain reproducible working
Table 1
LC-EC conditions
Column
Spherisorb ODS2, 100x4.6 mm,
3 µm
Sample
1.5 ml/min
acetonitrile
50 mM HAc/NaAc, pH 4.0, 35%
injection
100 - 1000 nM phenols, 20 µl
Temperature
30 °C
Flow rate
Mobile phase
are between 25 and 220 ng/l,
except for DNP (0.9 µg/l), TCP
(0.95 µg/l) and PCP (6 µg/ml).
In the 70’s the US Environmental Protection Agency
(EPA) made a list of the eleven most important
phenol contaminants as priority pollutants.
Phenols
- www.myantec.com
- The Netherlands - [email protected]
12 - 2382 NV - Zoeterwoude
- www.myantec.com
Antec Leyden B.V. - Industrieweg
MD 21076 - USA - [email protected]
Connelley Drive - Hanover
Antec (USA) - Suite 110 - 7502
Proven
analyzers
to guarantee
your performance
Proven
analyzers
to guarantee
your performance
ENVIRONMENTAL
ALEXYS™ Analyzers
Fig. 5. Schematic representation of mobile phase bottle with Helium
sparging assembly.
Take the following precautions to assure carbonate-free mobile
phases
se
Fru
Fig. 3. ALEXYS 100 ‘Carbohydrates I’ isocratic, including column, flow cell
and LC connection kit.
cto se
se
se
Ma
lto
se
X Axis Title
Glu
co
Fru se
cto
La se
ct
&
Su
cr
pulsed amperometric detection, “fingerprints” of
ANALYSIS OF CARBOHYDRATES
ANION-EXCHANGE CHROMATOGRAPHY AND PULSED AM-PEROMETRIC DETECTION
Configurations
Potential (V)
With this high-pressure gradient analyzer based on
INTROD
UCTION
Carbohydrates
not only pro• Carbohydrates
vide the most
in food industry
easily accessi
• Analytes
and fields in
ble energy
of simple monolife science
source for
s
or disaccharides
our
• Analytes
body, they
of oligosaccharides
also play an
(Maltodextrin)
• Analytes
important role in
of polysaccharides
many physiol
(starch, cellulos
• Analytes
ogical process
of glycoproteins
e)
es. Some examples of these
processes
are
intercellular
Due to the
recognition,
poor detection 220_002 #03
limits of RI,
ferred detection
infection process
EC has become
method in
es, and
trace analysis
the preChromatography
ANION-EXCHANGE
CHROMATOGRAPHY AND PULSED AM-PEROMETRIC DETECTION
certain types
. Anion-E
at
high
xchange
pH
of cancer.
amperometric
in combina
The
tion with pulsed
detection (PAD)
determination
analytical
is nowaday
of carbohy
method available
s the most
sensitive
drates is of
In this applicati
for carbohy
drate analysis
interest to
Page : 2/10
on note we
the
[1,2].
present a
solution for
food industry
simple and
the determin
Page : 2/10
sensitive system
and many
ation of sugars
food products
fields
and oligosac
in life science
based on the
charides in
ALEXYS 100
s.
'Carbohydrate'
Method
Analytes of
systems.
interest include
simple monoAnion Exchang
or disacch
e Chromatography
abehaviour
of the carbohydrates can be controlled by
> 12) carbohy The retention
rides (such
- Under
as glucose
drates can
alkaline and sodium acetate in the
be
E2 (Oxidation Au)
the concentration
of sodium hydroxidecondition
and
Exchange
s (pH
sucrose), oligosa
Chromatography separated by means
of Anionmobile phase.
An increase of the sodium
hydroxide concentration
ccharides
pKa values
. Carbohy
drates
ranging between
(Maltodextrin),
weak of
has a dual
effect on the are
retention
carbohydrates.
The
acids
either complete [OH-]
12 and
polysacchawith
14.
At
lyincrease
or partially
E1 (Determination)
pH they
rides (starch,
in ionic
strength of high
the eluent
causes
a decrease in
value. Due
will be
ionised
depending
cellulose) and
to the extreme
onpH
retention,
the higher
willpKa
increase the degree of
their
alkalinewhile
anion-exchange analyte
glycoproteins.
conditions
columns
only polymer
dissociation
an increase
in analyte
areresulting
The retention
suitable in
tdelay
tsample
ic retention. If the pH
Numerous
for carbohy
time> of
pKa
(full dissociation)
then the ionic
dominate the
carbohy
dratestrength
analytical techpKa value
separatiwill
drates is inversely
on.
and increase
niques for
separation
process and
the retention
decreases.
correlate
s significa
elution order
carbohydrate
d with This is illustrated
ntly with molecula
E3 (Reduction Au)
of carbohy
analin Fig.drates
1A.
r weight. The
ysis have been
is usually as
on such anion-ex
follows: sugar
developed
change columns
alcohols elute
di-, tri-, and
over the years
Time (s)
higher oligosac
first, followed
[6]. The polar,
charides.
by
A. 50 m M NaO
H -mono-,
1 m M NaO Ac
non-volatile,
B. 30 m M NaO H - 1 m M NaO Ac
water-soluble
Fig. 2. Three-step potential waveform for the pulsed
amperometric
C. 15 m M NaO H - 1 m M NaO Ac
carbohydrates
detection of Carbohydrates.
are not easily
A
re-solved.
Also detectio
For that reason Pulsed Amperometric Detection (PAD) with a gold
n
of
these non-chr
(Au) working electrode has been developed in order to enable
omophoric
B
compounds
frequent (typically 1 - 2 Hz) renewal of the working electrode (WE)
is difficult.
1 2
Carsurface [ , ]. The repeating three-step potential waveform is
bohydrates
hardly have
schematically shown in Fig. 2. The three potentials E1, E2 and E3
any
UV absorpt
ion and therefor
are
applied
to
the
Au
WE
for
specified
times
t
,
t
and t3, respec1
2
C
e
either refractiv
tively. E1 is the actual detection potential. During t1 the carbohye index (RI)
or
drates are oxidized at the Au electrode surface resulting in a
electrochemical
0
5
10
15
20
25
30
detection
specific oxidation current. This oxidation current is sampled during
(EC) is used
M inutes
for analysis
a small time period tsample at the end of t1. The delay time tdelay
.
®
11
ALEXYS™ Analyzers
ALEXYS™ analyzers line-up
27 major configurations on display
Order no.
Standard configurations
180.0035A
180.0036A
180.0037A
ALEXYS Cool
ALEXYS Basic
ALEXYS Cool, Micro
Neuroscience
180.0088A
180.0070A
180.0067A
180.0068A
180.0082A
180.0083A
180.0081A
180.0091A
180.0092A
OFF-LINE ANALYZERS FOR MICRODIALYSATES AND BRAIN TISSUE
ALEXYS Monoamines Analyzer
ALEXYS GABA and Glutamate Analyzer
ALEXYS Acetylcholine Analyzer
ALEXYS Disulfides Analyzer
ON-LINE ANALYZERS FOR MICRODIALYSATES
ALEXYS OMD High Throughput
ALEXYS OMD Time Resolution
ALEXYS OMD Monoamines Analyzer
MODULAR ALEXYS CONFIGURATIONS
ALEXYS Flex SCC
ALEXYS Flex DCC
Pharmaceutical & Biotech
180.0050A
180.0058A
180.0056A
180.0059A
180.0086A
180.0068A
ALEXYS
ALEXYS
ALEXYS
ALEXYS
ALEXYS
ALEXYS
Aminoglycosides Analyzer
Kanamycin/Amikacin Analyzer
Gentamicin Analyzer
Spectinomycin/Lincomycin Analyzer
Azithromycin Analyzer, USP
Disulfides Analyzer
Clinical & Diagnostics
ALEXYS Clinical Analyzer
180.0068A
180.0095A
180.0078A
ALEXYS Disulfides Analyzer
ALEXYS Iodide Analyzer
ALEXYS Vitamins Analyzer
Food & Beverage
180.0090A
180.0094A
180.0054A
180.0095A
ALEXYS
ALEXYS
ALEXYS
ALEXYS
Bisphenol A Analyzer
Polyphenols Analyzer
Carbohydrates Analyzer
Iodide Analyzer
Environmental
180.0094A
®
12
190.0035A
ALEXYS Polyphenol Analyzer
ALEXYS™ Analyzers
Analytes
Documentation
Monoamines & metabolites
Gaba/Glutamate
Choline/Acetylcholine
Glutathione/Disulfides
213_017, 221_004
213_019, 221_009
213_022
211_004
221_011, 221_012
221_011, 221_012
221_011, 221_012
Neurotransmitters, add kit to this configuration
Neurotransmitters, add kit to this configuration
All neurotranmitters applications
All neurotranmitters applications
Neomycin/Framycetin/Tobramycin/Spectinomycin
Amikacin and Kanamycin
Gentamicin
Spectinomycin and Lincomycin
Azithromycin
Glutathione and Disulfides
217_010, 217_014, 217_018, 221_005
217_015
217_013
217_019
217_011, 217_017
211_004
Catecholamines
Serotonin
Metanephrines
VMA, HVA, 5-HIAA
Homocysteine
Iodide
Vitamins
214_001,
214_003,
214_005
214_006
211_003,
212_003
218_005,
Bisphenol A (from polycarbonate bottles)
Catechin, Flavonoids and other Polyphenols
Carbohydrates
Iodide
216_005
216_006, 221_019
220_002, 221_007
212_004, 221_022
Phenols in water
216_001, 216_002
214_002
214_004
221_003
218_001
Basic systems extendable with kits
®
13
Detectors
Part no.
Description
171.0035
171.0038
171.0039
171.0040
171.0035 MD
171.0038 MD
DECADE
DECADE
DECADE
DECADE
DECADE
DECADE
II
II
II
II
II
II
w/o
w/o
w/o
w/o
w/o
w/o
flow
flow
flow
flow
flow
flow
cell,
cell,
cell,
cell,
cell,
cell,
single cell control
dual cell control
triple cell control
quadruple cell control
single cell control
dual cell control
Specifications DECADE II
General specifications
Power
Operating modes
Potential range
Potential range “MD”
Output
Offset
Event marker
Auto zero
RS232C
Injector sensor
Oven
Diagnostics
Service mode
Configuration
Firmware
Environmental
DC mode
Ranges
Filter (cut off)
Noise
PULSE mode
Range
Filter (cut off)
Pulse times
Sample times
SCAN mode
Range
Scan rate
Cycle
10 pA – 200 μA in 1, 2, 5 steps
Advanced Digital Filter, 0.5 - 0.001 Hz, 1, 2, 5 steps
Better than 2 pA with a dummy cell (load of 300 MOhm and
0.5 μF), filter off, Ec +800mV and temperature of 30°C.
10 nA – 200 μA in 1, 2, 5 steps
t1: 100 - 2000 ms; t2: 0 - 2000 ms; t3: 0 - 2000 ms
in 10 ms steps
20, 40, 60, 80 and 100 ms
10 nA – 200 μA in 1, 2, 5 steps
1 - 50 mV/s in 1, 2, 5 steps
half, full or continuous
AUTO mode
DC mode (5 files) and pulse mode (4 files), cycle time, number of cycles and oven
temperature. Time-based control of 50 time points as to range, filter, output contacts
(2 TTL, 2 relays), auto zero, board ID, offset, valve position (if present) and cell potential.
Rear panel I/O connections
Mains, Output, 2 Connectors 15 pins (A, B), manual valve (C), RS232C connector
Physical specifications
Dimensions
®
Weight
14
100-240 VAC, 50/60 Hz, 260 VA max., auto-sensing
DC, PAD and Scan
Between +2.00 and -2.00 V in 10 mV increments
Between +2.50 and -2.50 V , activation tool MD
Between +1 and -1 V or between +10 and -10 V
(20 bit D/A converter)
Between +50% and -50% of max. output voltage, 5% steps
Pulse of 10% of max. output
Triggered by keyboard, rear panel TTL, or RS232C control
Instrument control, data acquisition at 1, 2, 5 and 10 Hz
Starts system clock at injection
Height 37 cm, from 7°C above ambient to 45°C, accuracy
0.5°C, stability 0.1°C; accommodates column and flow cell(s)
LCD screen, keyboard and noise (internal dummy cell)
System settings and calibration parameters
Menu for system customization and optimization
Upgradeable via PC (RS232), flash technology
Operating temperature: 4 – 40°C, RH 20 to 80%,
non-condensing
44 (D) x 22 (W) x 44 (H) cm = 17.3” (D) x 8.7” (W) x 17.3”
(H)
14.6 kg (32 lbs) without flow cell and column
Detectors
DECADE II
Digital electrochemical amperometric detector
• Accurate
•Versatile
•Sensitive
Accurate
Both chromatography and EC detector responses are
strongly affected by temperature fluctuations. It is a
prerequisite for trace analysis to have a shielded and
thermostatted environment for both the flow cell and
column (fig 1).
Versatile
The DECADE II can control up to 4 flow cells. The column/
flowcell combination can be used in parallel, but also
serial arrangements are possible. To facilitate flow cells
with Magic Diamond™ electrodes the DECADE II has an
equipped with DC, Pulse and Scan mode.
Sensitive
The DECADE II demonstrates superiority in EC detection
and sets a new standard in design and performance. For
electrochemical detection applications, sensitivity is crucial.
The DECADE II with a newly developed Advanced Digital
Filtering (ADF), is breaking new records in detection limits.
A DECADE ahead
activation pulse button. In addition, the DECADE II is
Fig. 1. Baseline without temperature control (A)
and baseline of DECADE II with temperature control (B).
®
15
Detectors
Part no.
Description
174.0035
DECADE II SDC w/o flow cell
Specifications DECADE II SDC
Offset
Event marker
Auto zero
RS232C
Oven
Environmental
DC mode ranges
Environmental
Dimensions
Weight
44 (D) x 22 (W) x 44 (H) cm = 17.3” (D) x 8.7” (W) x 17.3” (H)
14 kg (32 lbs) without flow cell and column
Auto mode
100-240 VAC, 50/60 Hz, 260 VA max., auto-sensing
DC
Between +2.00 and - 2.00 V in 10 mV increments
Between +1 and - 1 V or between +10 and -10 V (20 bit D/A
converter)
Between +50% and - 50% of max. output voltage, 5% steps
Pulse of 10% of max. output
Triggered by rear panel TTL or RS232C control
Instrument control, data acquisition at 1, 2, 5 and 10 Hz
Height 37 cm, from 7°C above ambient to 45°C, accuracy 0.5°C,
stability 0.1°C; accommodates column and flow cell(s)
Operating temperature: 4 – 40°C, RH 20 to 80%,
non-condensing
10 pA – 200 μA in 1, 2, 5 steps
Better than 2 pA with dummy cell in 1 nA/V range, filter off, Ec
+800mV and temperature of 30°C.
DC mode (5 files), cycle time, number of cycles and oven temperature. Time-based control of 50 time points as to range, filter,
output contacts (2 TTL, 2 relays), auto zero, board ID, offset,
valve position (if present) and cell potential.
Operating temperature: 4 – 40°C, RH 20 to 80%, non-condensing
Filter (ADF)
Noise
®
16
Power
Operating mode
Potential range
Output
Detectors
DECADE II SDC
Plain performance
• SDC: Single Cell Control, DC mode only
• ADF: Advanced Digital Filter for noise suppression
• controlled by (free) PC software
The DECADE II SDC has the same performance as the
famous DECADE II. It has an integrated oven with Faraday
cage and is equipped with the superior ADF. The DECADE
II SDC can control one flow cell and runs in DC mode only.
Without the presence of front panel control, the unit is
operated by PC control using our data acquisition software
or a free version of Dialogue™ software. The DECADE II
SDC is compatible with all Antec flow cells. An electrically
actuated injector or a manual valve is optional.
Integrated oven with Faraday cage
Comparison ADF and standard noise filter
1.000
Signal
height
0.750
ADF (Antec)
Standard filter
peak
frequency
noise frequency
0.500
Noise
0.250
Noise
Using ADF results in a factor
10 to 100 times better S/N Ratio.
0.000
0.010
0.100
1.000
10.000
®
f(Hz)
17
Autosamplers
Part no.
Description
181.0037
181.0038
Autosampler AS 100, cool, micro, 6-PV
Autosampler AS 100, cool, micro, 10-PV
Specifications Autosampler AS 100
Power
Operating conditions
Sample capacity
Vial dimensions
115 or 230 VAC ± 10%, 50/60 Hz, 250 VA max
10 - 35°C, 20 - 80% RH, non-condensing
160, 96, 72 or 32 vials (20, 12, 9 or 4 per segment)
7, 12, 15, or 22 mm vial diameter min/max height: 32/46 mm
(including cap) micro vial inserts for micro dialysis
Dispenser
Syringe: 100, 250 (standard), 500, 1000 μL, or 10 mL
SS316, PTFE, TEFZEL, VESPEL, glass, PEEK, silica
Wetted parts
Loop volume
Injection
Analytical performance
Injection programming
Fool loop
Partial loop fill
RSD < 0.3% (all specs for capped and sealed vials)
µL Pick-up
Carry-over
RSD < 1.0%, injection volumes > 5 μL, with headspace
pressure on the vial
< 0.01% with programmable needle wash
Methods
Full loop, partial loop and μL pick-up, user defined programming
Volumes
1 μL - 1 mL (method dependent)
Vial, volume, repetitions: freely programmable in data
acquisition software
Programmable needle wash
Injections
Wash
Connections
Outputs
Inputs
PC control
Options
RSD < 0.5%, injection volumes > 5 μL, with headspace
pressure on the vial and 30 μL pre-flush with air segment
Marker TTL and contact closures: inject, vial, labelled vial,
stop I/O; programmable: 4 TTL, 4 relays contacts no/nc,
2 relays contacts no/nc, 4 TTL time based; alarm
Next injection, next vial, freeze, stop, 4 TTL
RS232C, full control with data acquistion software
Sample cooling
4 - 40°C; accuracy: ± 2°C capacity: ambient - 20°C
(measured on cooling tray)
Micro option
Optimized for small injection volumes > 1 μL; with 5.3 μL
needle, 20 μL loop and 100 μL syringe volume, μ-bore valve
Dimensions
54 (D) x 28 (W) x 44 (H) cm
21.3” (D) x 11.0” (W) x 17.3” (H)
22 kg (48 lbs), 30 kg (66 lbs) incl. cooling option
®
Weight
18
5 μL - 1 mL (1, 2, 5 steps), 100 μL standard
Pre-puncturing septa/caps with air needle, dual needle design,
vial & height detection by vial sensor
Autosamplers
AUTOSAMPLER AS 100
Fast, accurate & reproducible
• automated derivatization, dilution, pH adjustments
• injection volume < 10 µL without sample loss
• Sample pick-up down to 1µL
• Cooled sample tray
Flexible vial choice
The AS 100 has impressive records in injection precision
and sample handling. It can be equipped with sample tray
cooling to protect labile samples against degradation, which
can be crucial in various LC-ECD applications. Highest
injection performance is provided in the flushed and partial
loop fill mode. If no sample loss can be accepted, e.g. in
case of µ-dialysate sampling, the custom injection mode
can be used.
The coaxial design of the sample needle inside a prepuncture needle allows for penetration through various
septa without damaging the sample needle. The AS 100
of any volume from any vial into any destination vial.
Pre-column derivatization, dilution and pH adjustment can
also be automated. Programmable wash steps decrease
carry-over to an absolute minimum.
Programming is straightforward and intuitive via the data
acquisition software.
Customized injection program
to minimize sample loss
features mixing routines enabling aspiration and dispensing
®
19
Autosamplers
Part no.
Description
191.0035
191.0036
191.0037
191.0038
191.0039
Autosampler
Autosampler
Autosampler
Autosampler
Autosampler
AS
AS
AS
AS
AS
110,
110,
110,
110,
110,
standard, cool, 6-PV
standard, 6-PV
cool, micro, 6-PV
cool, micro, 10-PV
micro, 6-PV
Specifications Autosampler AS 110
Power
Sample capacity
Vial dimensions
Dispenser
Wetted parts
Loop volume
Sampling technology
Analytical performance
Injection programming
20
µL Pick-up
Carry-over
RSD < 1.0 %, injection volumes = 10 μL
< 0.05 % with programmable needle wash
RSD < 0.5 %, injection volumes = 10 μL
Methods
Full loop, partial loop fill and μL pick-up, user defined programming
Volumes
1 μL – 5000 μL (with 1 μL incr.), depending on system settings
Programmable, between injections or vials
Outputs
Sample cooling
Bio-compatible
®
1 μL – 5 mL (1,2,5 steps) and 10 mL (prep. option)
PASA™, pressure-assisted sample aspiration no sample syringe
contact, fixed concentric air/sample needle pair. Injection cycle
time < 20 s, depending on settings
RSD < 0.3 % (all specs for capped and sealed vials)
Inputs
PC control
Options
Syringe: 500 μL (standard) or 2.5 mL (prep. option)
SS316, PTFE, TEFZEL, VESPEL, Glass, Teflon. Optional: PEEK
Fool loop
Partial loop fill
Wash
Connections
95 - 240 Volt AC ± 10%; 50 - 60 Hz; 200VA
10 - 40°C , 20-80% RH
Well Plates: 2x 96-well high/low and 384-well low formats,
according to SBS standards. Vials: 2 trays of 48 (1.5 mL) or 12
vials (10 mL)
Max. plate/vial height: 47 mm (incl. septa or cap-mat)
1 programmable relay output, programmable as inject marker
(default), aux, alarm
2 programmable TTL inputs, next injection , freeze, stop
RS232C, control via acquisition software
Built-in Peltier cooling, non-condensing, range: 4°C
to ambient - 3°C
Inert sample needle (Silco steel) and bio-compatible valve (PEEK)
Prep. kit
2.5 mL syringe, prep valve, 10 mL sample loop, LSV needle
and sample tray for 10 mL vials
Dimensions
300 (W) x 575 (D) x 360 (H) cm (with cooling option)
Weight
19 kg (42 lbs), 21 kg (46 lbs) with cooling
Max. weight on top
65 kg (143 lbs)
Autosamplers
AUTOSAMPLER AS 110
Fast, accurate & reproducible
• unique needle concept
• sample pick-up down to 1µL
• cooled sample tray
• well plates or vials
The AS 110 is a new generation autosampler. It is a fast,
robust and easy to use autosampler without compromising
accuracy and reproducibility. The AS 110 includes
indispensable features for the highest injection
performance. It has a dual needle with positive headspace
pressure, extensive wash routines for minimal carry over
and 3 injection modes including µL pick-up mode for zero
sample loss.
The AS 110 can handle sample volumes down to 1 µL
accurately and reproducibly. The sample tray offers temperature control down to 4°C. The fast needle movements
result in unsurpassed speeds of less than 30s for a full
high throughput routine analysis.
The AS110 cool, micro, 10 PV is equipped with an unique
stainless steel needle with a total dead volume of 2.4 µl
and a micro bore flow path. This allows you to program
small sample injections without peak broadening and loss
of sample.
Perfect fit for high throughput,
routine analysis & small samples
cycle. These features make the AS 110 a perfect fit for
®
21
Pumps
Part no.
Description
193.0035
Pump LC 110
Specifications HPLC Pump LC110
Delivery system
Power
Flow rate
Flow rate accuracy
Ripple
Displacement volume
Pressure protection
Operating modes
Instrument control
GLP features
Dimensions
®
Weight
22
Dual-piston pump with main and auxiliary piston in a serial
configuration, automatic piston flush.
Auto-sensing, 90 - 260V, 47 - 63 Hz, 70 VA
10 - 40°C, 20 – 80% RH, non-condensing
0.001 - 9.999 mL/min, 0 - 40 MPa
< 1.5 % at 2 - 20% FS
Residual pressure pulsation < 3% (water,12 MPa)
Primary piston 21.4 μL secondary piston 9.7 μL
Pmin: 0 (off) - 40 MPa, switch-off in 30 s.
Pmax: 0.1 - 40 MPa, switch-off time < 1 s
Isocratic, gradient (high pressure, up to 4 eluents)
Numerical keypad & LCD, RS232C full digital control,
REMOTE connector (stop, flow, pressure, error)
Report of number of pump cycles, working time, delivered
volume and service information
412 (D) x 226 (W) x 135 (H) mm 16.2 (D) x 8.9 (W) x
5.3 (H) inch
5.3 kg (11.7 lbs)
Pumps
PUMP LC 110
Superior performance LC pump for ECD
•Optimized for ECD applications
• ultra low pulsation
• high accuracy at low µL flow rates
•automated piston wash
The LC 110 pump is designed for lowest pulsation in
combination with electrochemical detection.
Consequently under optimized conditions no noise is
detectable from the piston stroke.
The LC 110 pump head is available with stainless steel,
titanium or ceramic inlays. These various pump head
inlay materials meet the requirements of just about
every application. High pressure gradient as well as
flow rate programming is supported. The control
software supports the programming of a quaternary
high pressure gradient system.
Baseline noise (ECD)
Dual piston design
with active pulsation dampening
The LC 110 pump features a dual piston design and an
automated piston wash to remove traces of salt and
other contaminants from the backside of the pistons,
increasing the lifetime of the pump seals. The pump
head can deliver flow rates in the low μL/min range up
to 10 mL/min.
Fig 1. Baseline in ECD with LC110 (A)
compared to another brand (B).
®
23
Bottle rack
Tool rack
Part no.
Description
184.0040
184.0035
Organizer OR 110
Organizer rack OR 100
Pump rack
Specifications Organizer OR 110
Pulse damper
100 – 240 VAC, 50/60 Hz, 20 VA degasser only
10 – 35°C, 20 – 80% RH, non-condensing
8 outlets: 100 VAC (max 800 VA total) – 240 VAC (max
1900 VA total), max 300 VAC per outlet Degasser 2 channels,
volume 480 μL/channel, capacity < 2 ppm O2 at 1 mL/min,
max flow rate 10 mL/min LED for power, start-up and vacuum
level wetted materials: PEEK, PTFE, Teflon AF
Membrane type, dead volume 0.9 mL at max pressure
41.4 MPa (6,000 PSI wetted materials: 316 SS and FEP
Dimensions
35 (D) x 9 (W) x 44 (H) cm
Weight
10.3 kg (22.7 lbs) with 1 pulse damper
Specifications Bottle rack BR 110
®
24
Power
Power outlets
Dimensions
46 (D) x 30 (W) x 17 (H) cm
Weight
6 kg (13 lbs)
Organizer And Racks
Degasser, pulse damper and stackable, compact
• mains power distributor for ALEXYS™
• dual channel degasser
• integrated pulse damper
• Stackable
• Leakage drain
The compact Organizer OR 110 includes an extremely efficient
degasser and pulse damper. It can be upgraded with a second
pulse damper. The power supply for all ALEXYS™ components
comes from the OR 110 avoiding grounding loops.
Two independent vacuum channels
• Dead volume 480 ul per channel
• Continuous vacuum degassing by means
of a variable RPM vacuum pump
• Teflon AF
(tool rack), just to organize your system.
An extremely efficient degasser
– stackable racks
The PR 110 (pump rack), SR 110 (solvent rack) and TR 110
®
25
Flow cells
Why amperometry?
nanofibers
amperometric
coulometric
• low noise
• best S/N ratio
log
b
c
log WE area
110.4105
110.4205
110.4305
110.4115
110.4215
110.4315
111.4105
®
26
• highest signal
• poor S/N ratio
S
Part no.
coulometry
• highest noise
a
N
amperometry
• about 5 – 10 % conversion
d
S/N
Why amperometry?
Figure from D. M. Morgan and S. G. Weber
Anal. Chem., 56, 1984, p. 2560
Description
Glassy Carbon working electrode
VT-03 flow cell, 3 mm GC WE, sb REF
VT-03 flow cell, 3 mm GC WE, ISAAC
VT-03 flow cell, 3 mm GC WE, HyREF
VT-03 flow cell, 2 mm GC WE, sb REF
VT-03 flow cell, 2 mm GC WE, ISAAC
VT-03 flow cell, 2 mm GC WE, HyREF
µ-VT-03 flow cell, 0.7 mm GC WE, sb REF
111.4205
μ-VT-03 flow cell, 0.7 mm GC WE, ISAAC
111.4305
μ-VT-03 flow cell, 0.7 mm GC WE, HyREF
110.4120
110.4220
110.4320
110.4125
110.4225
110.4325
110.4130
110.4230
110.4330
110.4135
110.4235
110.4335
Metal working electrode
VT-03 flow cell, 3 mm Pt WE, sb REF
VT-03 flow cell, 3 mm Pt WE, ISAAC
VT-03 flow cell, 3 mm Pt WE, HyREF
VT-03 flow cell, 3 mm Au WE, sb REF
VT-03 flow cell, 3 mm Au WE, ISAAC
VT-03 flow cell, 3 mm Au WE, HyREF
VT-03 flow cell, 2 mm Ag WE, sb REF
VT-03 flow cell, 2 mm Ag WE, ISAAC
VT-03 flow cell, 2 mm Ag WE, HyREF
VT-03 flow cell, 3 mm Cu WE, sb REF
VT-03 flow cell, 3 mm Cu WE, ISAAC
VT-03 flow cell, 3 mm Cu WE, HyREF
ISAAC reference electrode
In Situ Ag/AgCl reference electrode for
VT-03 electrochemical flow cell
•Ultimate robustness
•No trapping of air bubbles
•Easy ‘plug and play’ installation
Reference electrode selection
•Standard: ISAAC
•Traditional or restrictions: ‘good old’ salt-bridge Ag/AgCl
•Extreme conditions: Hy-REF
Flow cells
VT-03 electrochemical flow cell
Highest sensitivity electrochemical wall-jet flow cell
•Quick stabilization
•Long lifetime (5 year warranty)
•Down to 5 nL working volume
•Easy maintenance
After extensive testing, it was determined that the confined wall-jet configuration gave excellent results. In
addition, we found that the qualilty of the material used
and the finishing process of the working electrode greatly
affected the performance of the EC detector. While
competitor flow cells deteriorate during use, the
performance of the VT-03 flow cell increases with use.
The VT-03 requires unusually short stabilization times:
Wall-jet configuration
for excellent results – short stabilization times
trace analysis within half an our after start up can be
expected. We have so much confidence in our VT-03 flow
cell that we warrant the glassy carbon flow cell for a
period of 5 years.
such as glassy carbon, platinum, gold, silver and copper.
A selection of working electrodes of 3, 2 and 0.7 mm diameter together with salt bridge, ISAAC and HyREF
reference electrodes ensures compatibility with any type of
HPLC analysis.
The VT-03 is available with working electrode materials
®
27
Flow cells
REF
reference electrode
reservoir
open channel
to REF reservoir
auxiliary electrode
inlet block
50 µm spacer
working
electrode
disk
50 µm spacer
Working electrode
assembly
Part no.
Description
102.4305M
102.4305
102.4320
102.4325
102.4330
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
MD HyREF
GC HyREF
Pt HyREF
Au HyREF
Ag HyREF
102.4105
102.4120
102.4125
102.4130
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
GC sb
Pt sb
Au sb
Ag sb
102.4205
102.4220
102.4225
102.4230
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
GC ISAAC
Pt ISAAC
Au ISAAC
Ag ISAAC
Part no.
Description
102.5007
102.5022
102.5027
102.5032
102.5037
102.5050
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
FLEXCELL
electrode
swivel nut
HyREF reference electrode
Maintenance-free reference electrode,
easy ‘plug and play’ installation
• pH resistant (1 - 14)
•Compatible with 100% modifier
®
28
WE
WE
WE
WE
WE
WE
disc
disc
disc
disc
disc
disc
•Inert
Reference electrode selection
GC (glassy carbon)
Pt (platinum)
Au (gold)
Ag (silver)
Cu (copper)
MD (magic diamond)
•Standard: ISAAC
•Traditional or restrictions: ‘good old’ salt-bridge Ag/AgCl
•Extreme conditions: Hy-REF
Flow cells
FLEXCELL
The most versatile flow cell for LC-ECD
• exchangeable working electrode
•Working Electrodes: GC, Pt, Au, Ag, Cu, MD
• low cost of ownership
The name FLEXCELL is chosen since it emphasizes the
versatility and serviceability of this thin-layer flow cell
in the Antec program. With its unrivaled design working
electrodes can be serviced or replaced in a few minutes.
The low cost of ownership is attributed to the replacement
of only the working electrode disc. The same holds for
switching electrode material for different applications
using the same flow cell. This makes the FLEXCELL
suitable for all sorts of electrochemical analyses like the
usual biogenic amines (glassy carbon), but also
carbohydrates (gold or copper), peroxides (platinum),
halides (silver), sulfides (Magic Diamond) etc.
The FLEXCELL properties can be fully exploited in
applications using metal working electrodes. A typical
example is the pulsed amperometric detection of
carbohydrates, where consumption of the gold working
electrode occurs. Renewal of the gold electrode is done in
a few minutes when the erosion has become unacceptable.
The total cell volume is less than 1 µL, which makes the
FLEXCELL suitable as an electrochemical reactor in-line
with other detectors such as a second EC flow cell.
Flexibility replaceable working
and reference electrodes
Applications
®
29
Kits, columns and valves
Valves
250.0012
Valco manual injector
250.0017C
Repl. valve for Valco electr inj, 6-port
250.0013
Valco manual injector, micro
250.0022
Valco electrical injector, 4-port, micro
250.0014
OMD valve option Valco, 14-port
250.0025
Valco electrical injector, 10-port
250.0017
Valco electrical injector, 6-port
250.0026
OMD valve option Valco, 6-port
250.0017A
Seal for Valco electr injector, 6-port
250.0030
DECADE II valve mounting panel kit
250.0017B
Stator for Valco electr injector, 6-port
Columns
®
30
250.1050
HPLC column for OQ
250.1114
ALF-115 column, 150x1.0mm, 3um C18
250.1068
ALA-510 column, 100x4.6mm, 5um C18
250.1116
250.1070
ALA-525 column, 250x4.6mm, 5um C18
250.1118
250.1071
ALA-105 column, 50x1mm, 3um C18
250.1120
250.1072
ALA-115 column, 150x1mm, 3um C18
250.1122
250.1080
ALC-525 column, 250x4.6mm, 7um
250.1125
250.1082
ALC guard column starter kit
250.1126
ALG-515 column, 150x4.6mm, 3um
250.1084
ALC guard column repl. cartr. 5/pk
250.1128
ALG guard column inserts, 3pk
250.1094
ALB-215 column, 150x2.1mm, 3um C18
250.1130
ALG guard column holder
250.1095
250.1132
ALH precolumn cartridges 5x1mm, 5pk
250.1096
250.1134
ALH precolumn holder 5mm
250.1097
250.1500
Column guard holder 4.0-4.6mm
250.1100
ALD-510 column, 100x4.6mm, 7um AEX
250.1502
Column guard holder 2.1-3.0mm
250.1101
ALD guard column starter kit
250.1504
Column guard holder 1.0mm
250.1102
ALD-515 column, 150x4.6mm, 7um AEX
250.1516
ALF 3um drop-in cartridge 10x4.0mm,4pcs
250.1104
ALE-315 column, 150x3.0mm, 3um C18
250.1518
250.1105
ALE-310 column, 100x3.0mm, 3um C18
250.1520
250.1110
250.1522
ALA 5um drop-in cartridge 10x4.6mm,4pcs
250.1112
Kits, columns and valves
columns, KITS and valves
Accessories for trouble free analysis
•Standardization
•Easy & fast installation
•Reduction of the total installation time
•Optimal performance
For most applications the choice of column, chemicals and
other accessories for sample pre-treatment are crucial for
trouble free analysis. Antec’s system solutions include all
necessary parts and chemicals to make your analysis work.
To obtain superior performance relevant system details are
optimized. For example, connecting tubing is prefabricated,
sharply cut, and optimized in diameter and length to avoid
dead volume. All Antec columns are flushed extensively
removing all electrochemically active contaminants, and are
Schematic drawing LC connections
shipped ready for EC analysis.
®
31
Software
Device control
An extensive list of devices can be controlled by Clarity,
operational parameters are stored in method files and
chromatograms. It makes methods easily interchangeable
between systems.
Peak table and chromatogram
Users can easily edit acquired chromatograms visually in
the graph or through the integration table. The Result tables
(Results, Summary, Column Performance, etc.) can be
customized to display the data you want to see.
The Summary tab displays data from overlaid
chromatograms in one summary table.
Report options
A fully featured report editor allows reporting data on
customized templates.
Sample queue
The Sequence table is a user friendly interface to
compose a queue for automated injections. The status of
each row is indicated by the colored symbols.
Copy, paste, fill down, auto increment and time/date
fields for file title are available.
Description
195.0035
Clarity single instr. incl LC, AS mod.
195.0037
Clarity single instr. incl LC mod.
195.0C50
Clarity Single Instrument SW (1tb)
195.0C55
Clarity Single Instrument Add-on
195.0C59
Clarity Evaluation off line version
Tray menu
The AS 110 can be used with a large variety of well
plates and vials. Up to 4 specific reagent positions can
be defined.
Hardware requirements
Software requirements
The minimal PC hardware requirements for the installation
of the ROXY™ system in combination with the
Clarity™ data software are listed in the table below:
Operating system Windows XP/2000/Vista/7
Beside the minimum hardware requirements we strongly
recommend to have the following software installed:
The Windows operating system should be
updated with the latest Service Packs (SP).
Processor
> 1 GHz
RAM
> 512 MB
Monitor Resolution 1024x768, 64K
®
32
Part no.
Software
Use
Adobe PDF reader
Reading electronic versions of
the user manuals
Accessing the DataApex and
Antec support websites, for
updates of software, manuals,
troubleshooting or other info.
Statistical data analysis
Internet explorer 7.0 or
other browser +
internet connection
Hard disc
NTFS, >10GB free HD space
USB
2 free USB ports (USB 2.0)
MS Office (Excel®)
PCI
2 free full-size 32-bit PCI 2.0 slots
WinZIP or other archiver
Drives
DVD or CD-ROM
Extracting Clarity™
pre-configured system files
Software
Clarity
Clarity software for system control and data acquisition
• intuitive
•Modular software package
•Up to 4 independent timebases
•Multiple instrument control
•GLP, 21 CFR part 11 compliance
Clarity from DataApex is an advanced modular software
package for data acquisition, data processing and
instrument control. Up to four independent LC systems
each with up to 12 detectors can be simultaneously
connected. Optional ‘Control modules’ provide integrated
control of selected instruments, ‘Extensions’ provide
specialized functionality for PDA or GPC analysis. Its wide
range of data acquisition interfaces (A/D converters, LAN,
USB, and RS232) allows connection to virtually any
Instrument window
The Instrument window is the control center of the whole
process of data acquisition and evaluation.
Data analysis
The Calibration window has two main screens, a global
calibration table and dedicated tabs for each compound
(on the screenshot). It includes a table with concentration
levels matching the response with Amount and a calibration curve. Any point can be temporarily suspended in the
graph it will be marked with a circle instead of a cross.
chromatograph.
®
33
Software
®
34
PQ noise measurement
DECADE II Performance Qualification is implemented in
Dialogue™ software. It is an automated performance
qualification that results in a single page hardcopy PQ report. For PQ Noise measurement a dummy flow cell must
be installed.
Integrated scripting tool for OQ
An OQ script is a series of commands to set
parameters and read data from the detector. A script is
executed by the script manager in Dialogue™ software.
All results are printed in the OQ document which also
contains the noise and signal test data. For successful qualification a special OQ kit is available containing
cables and tools for testing.
Editor for time files, and transmitting to DECADE II:
Time file editor is a tool for writing and transmitting time
files to the detector.
Database for audit trial and user rights
An audit trail is available, keeping track of all events
initiated by registered users.
Part no.
Description
171.9005
171.9002
171.9007
Dialogue™ software, PQ version
Dialogue™ software, OQ/PQ version
Dialogue™ software, free version
Software
Dialogue™ software
Service and OQ/PQ tool for DECADE II
•OQ and PQ measurements
•Scanning voltammetry
•Simple data acquisition
•Noise measurement
Dialogue™ software for Windows is a multi-functional
program to control the DECADE II electrochemical detector.
Dialogue™ software is a tool for:
• controlling all operational parameters of DECADE II
• creating and transmitting time files to DECADE II
• controlling DECADE II IO ports for external devices
• control of DECADE II SDC
•OQ and PQ measurements
• scanning voltammetry
• noise measurement
Dialogue™ software is one software package containing
license dongle PQ/OQ functionality will be unlocked.
There are no tools for further data analysis, therefore
Dialogue™ software can not be considered as a fully
featured chromatography software package.
All relevant settings can be stored
on PC and loaded for later use
under identical conditions.
functionality for all different versions. Depending on the
®
35
Documentation
Application and technical notes
Proven performance
Hplc/ec
ecD
®
s fo
Ation
Applic Alysis
lc-ec
ce An
Artest
scien
neUro
eD
inD
terM
er MAs
ec/lc
/Ms
r
tHe sM
olites
e metab
and th
mines
Monoa
alin
en
dr
Nora
ine
)
Dopam
(5-HIAA
nin
ic acid on(DOEc/Ms
PAC)
Ec/Lc/Ms TEchnoLogy
Seroto
ole acet Focus
acid
cetic
ox yind
la
elec
trochemistry and mass spec trometry
5-hydr
ypheny
hydrox
ids
(HVA)
id
3,4- di
ino ac
ac
and am
anillic
es
ov
in
m
ho
ed am
rivatiz
e
OPA de
utamat
and Gl
ate (GABA)
GABA
obutyr
4- amin e (Glu)
focUS on ec ecD HPlc/ec tecHnology
at
ine
cholec/ms
elec trochemistry and liquid chromatography
Glutam
Acetyl
e and
Cholin
e (Ch)
in
ol
h)
Ch
oline (AC stress
ch
yl
Acet
ive
oxidat
rs for
sine
Marke
L-Tyro
®
®
3-nitro- T
ec/lc/ms thiols
PA
ec ecd
her
8- OH-D
ot
and
hione
Glutat
ROXY™ Potentriostat (DCC)
Electrochemistry/LC/MS a powerful tool for predicting metabolic processes
Invitrometaboliteidentification- Combining Electrochemistry
with LC/MS – EC/LC/MS a powerful
analytical
technique
feel
free
to contact
MPW-522
Reactive metabolites - Generationandidentificationofreactivemetabolites
using on-line LC/Electrochemistry/Mass Spectrometry
PMM - 479
On-Line Electrochemistry/LC/MS – a Powerful Analytical Technique
-
Life Sciences / Nucleotides & Nucleic Acids - LS/NNA - 01 Keynote:
Frontiers of MS in Genome Research
PWA - 081
Simulation of the Oxidative Metabolism of Caspase Binding
Radioligands by On-Line EC/LC/MS
PWA - 082
Simulation of the Oxidative Metabolism of the Calcium Channel
221-001
Attomole detection limits in mircro HPLC-ECD
Blocker (CCB) Verapamil Using On-Line EC/HPLC/ESI-MS
221-002Processes
Removal of solvent front and contamimants by a post-column valve
Electrochemical Simulation of Oxidation
Involving Nucleic Acids On-Line Monitored
ESI-MS and serotonin analysis
221-004 with
Dopamine
Predicting Metabolic Processes using
Electrochemistry/LC/MS
221-006
Ox-Red analysis for selectivity improvement
Implemantation of Wall-Jet Cells for
the Simulation
221-009
GABAofand Glutamate in microdialysate
Oxidative Phase-I Metabolism in on-line ES/HPLC/MS
Simulation of the oxidative phase I metabolism of tetrazepam
221-012
Multi-channel
on-line microdialysis
using online electrochemistry/HPLC/MS
tetrazepam
23
00351
221-014
Fast LC with simulation
electrochemical detection
Metabolic studies of tetrazepam based
on electrochemical
4 methods
in comparison to6x10
in vivo and in vitro
221-015
Fast LC of DA and 5-HT
23
00352
10
Interaction of thimerosal with proteins—ethylmercury
5
11
adduct formation2x10
of human serum albumin
8 and b-lactoglobulin A
6
12
221-011
1x106
2
TPI-240
sed mollis tincidunt, nisl sem posuere leo, non
Clarity software
HPLC/EC system
Antec (Leyden) B.V.
Antec (USA)
P.O. Box 111418
Lorem ipsum dolor sit amet, consectetur adipisc-
4
14
16
time [min]
Homocysteine
in plasma
7
10
12*
tetrazepam
Macrolide and Aminoglycoside
Antibiotics Analyser
1x10
6
12Azithromycin
14 16 time [min]
in
8
10
221-018
Palm Bay, FL 32911
Phone
(321) 473-4202
Fax
(321) 821-1938
[email protected]
2
4
6
8
10
12
14
0
2
4
6
Antec (Leyden) B.V.
Antec (USA)
Industrieweg 12
P.O. Box 111418
2382 NV Zoeterwoude
Palm Bay, FL 32911
The Netherlands
USA
Tel. +31 (0)71 581 33 33
Toll free (888) 572 0012
6
8
7
10
12
16 time [min]
14
Distributor
Fax +31 (0)71 581 33 34
Phone
(321) 473-4202
[email protected]
Fax
(321) 821-1938
www.myantec.com
[email protected]
Chloro- and Nitrophenols
216_00
THE FINEST
LC-EC
• FOOD
Important
part APPLICA
as neurotransmitters
TIONS FOR
& BEVERAG
E ANALYSI
S
INTROD
PROCES
• EVER
Tumors
of SED
the sympathoadrenal
system
UCT
mination of catecholamine
levels in the 24-hours urine
allows the detection of patho-
Technology folders
: 1/4
Application notes & Technical notes
220_002 #03
Page : 5/10
214_001 #01
H
Especially, during the isocratic analysis of carbohydrates with
weak eluents ([NaOH] < 50 mM) a gradual loss of retention is
observed due to the slow build up of interfering anions on the
column. If during the isocratic analysis of carbohydrates a loss of
retention is observed, regeneration of the column is necessary.
Regeneration of the column can be achieved by flushing the
column with a volume of 30 – 60 mL of carbonate-free 0.2 M
NaOH. After regeneration, the column should be allowed to reequilibrate again with mobile phase. Stabile retention times (RSD
< 0.4%) can be achieved again after flushing the column for 5
hour with eluent at a flow rate of 2 mL/min.
IN URINE
H
H
Fructose
HO
O H
H
OH
H
Dopamine
nA
O
Norepinephrine
12
14
min
1.2
1.0
0.8
0.6
The quantification of the catecholamines in the urine samples is
performed by means of a single-point calibration method using a
urine calibrator. The ClinCal® urine calibrator supplied in the
ClinRep® kit is a lyophilised urine sample with a known amount
of catecholamines. The urine calibrator should be processed via
the same sample preparation method as the urine samples. An
example chromatogram of a urine calibrator analysis is shown in
figure 1.
An internal standard method is used to compensate for recovery
losses during the sample preparation step. To every urine sample,
calibrator or control 30 μL of internal standard (IS) solution is
added. The IS response of the samples is compared to that of a
directly injected standard solution (ClinTest® standard) to determine the recovery. The sample response is then interpolated to
100% recovery to establish the real catecholamine concentration
in the urine samples.
Table 1
0.4
0.2
0
2
4
6
8
10
12
14
min
Fig. 2. Overlay of 6 chromatograms of 20uL injections of
ClinChek® control level I.
Table I. Calculated concentration of urine controls level I and II
(n=6). Concentration range specified by Recipe is given for reference (source: data sheet supplied with controls).
Specified conc (µg/l)
Min
Max
Calculated
RSD
Noradrenaline
conc (µg/l)
(%)
58.7
0.8
14
21
18.7
0.8
120
180
176.6
0.8
Noradrenaline
125
187
156.9
0.2
Adrenaline
29
43
35.1
1.5
Dopamine
186
278
236.9
0.5
Adrenaline
Dopamine
44
66
1.2
1.0
Flow cell
Column
ClinRep® Analytical column for catecholamines
in urine
0.4
0.2
Flow rate
1.0 mL/min
Sample
Mobile phase
20 µl, extracted with ClinRep® sample preparation columns
ClinRep® catecholamine buffer#
Temperature*
TD2 SDC 30°C (separation & detection),
E-cell
TAS110 4°C (sample cooling)
Range
500 mV (vs. Ag/AgCl sat’d)
I-cell
10 nA/V
ADF
Ca. 0.2 – 3.0 nA
Analysis time
Flow rate
Glucose
0.20
The control samples are lyophilised urine samples which have to
be processed in the same way as the urine samples. Both Control
I and Control II were analysed and the analyte concentrations
quantified using the ClinCal urine calibrator. For both urine controls level I and II the determined concentrations were within the
concentration ranges specified by Recipe on the urine control data
sheet (see table I).
0.1 Hz
15 minutes
#) mobile phase was recycled during experiments. *) minimum actual oven & tray
temperature which can be reached is dependent on ambient conditions.
4
2
3
4
5
6
8
10
12
B
C
D
E
5,0
5
10
15
20
An excellent linear detector response in the concentration
range between 20 nM and 10 µM was observed for the standard
mixture of sugars.
1,5
1,0
B
C
D
E
0,5
10
Glucose, r = 0.9993
Fructose, r = 0.9979
Sucrose, r = 0.9992
Lactose, r = 0.9995
0
5
10
15
20
7
min
0,1
Fig. 9. Reproducibility (n=23) of 20 μL injections of a standard mixture of 2
µM Glucose, 2 µM Lactose, 4 µM Fructose and 4 µM Sucrose in water.
Top: retention time, Bottom: peak area.
0,01
0,1
1
10
Concentration (μmol/L)
During the reproducibility experiments the mobile phase was
sparged continuously with Helium 5.0 to prevent the formation of
carbonate ions and subsequent loss in retention.
Fig. 8. Calibration plots of Glucose, Fructose, Sucrose and Lactose. Peak
area as function of concentration (concentration range 20 nmol/L - 10
µmol/L, 20 µL injections, n=10 per concentration). R determined via
weighted linear regression method.
Reproducibility
1
14
min
Fig. 3. Overlay of 10 chromatograms of 20uL injections of urine
sample B. Top-right: zoom in on NA and A peaks.
The RSD’s for all components were typically smaller then 4%.
Only for low concentrations of epinephrine, near the limit of quantitation, a RSD of 14 % was found.
For all urine samples, controls and calibrator recoveries typically
in the range of 85 – 95% were found, compared to a directly
injected standard. The concentration limit of detection (CLOD) for
the method was approximately 1 μg/L for all catecholamines. The
CLOD here is based on a 20 µL injection and defined as the concentration that gives a signal that is three times the peak-to-peak
Trademarks
- ALEXYS™, Dialogue™, FLEXCELL™ and VT03™ are trademarks of Antec (Leyden) B.V.
- Clarity™ is a trademark of Data Apex Company.
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- Excel® is a registered trademark of Microsoft Inc.
- Adobe® Acrobat® are registered trademarks of Adobe Systems Incorporated
25
Number of injections (n)
1
The relative standard deviations (RSD) of the retention times and
peak areas were determined of 23 consecutive injections of the
sugar mixture (see Fig. 9). The good reproducibility of the method
6
25
2,0
15
CLOD (nM)
10
15
15
10
0.05
0.0
2
Table 2
LC-EC conditions
Carbohydrate
Glucose
Fructose
Sucrose
Lactose
0.15
0.10
0.6
ALEXYS 110 LC-EC system with DECADE II
SDC (p/n 190.0035)
GC type flow cell with Ag/AgCl saltbridge REF
0.25
0.8
Control level II
HPLC
10
Fructose, Sucrose and Lactose dissolved in water.
cholamines using LC-EC.
nA
Control level I
Set-up
5
Fig. 7. Chromatogram of a 20 µL injection of 20 nM Glucose, Fructose,
Sucrose and Lactose in water.
15
urine samples and fast
& reliable
of urinary
cateTabel
2. Limit ofanalysis
Detection (LOD)
for Glucose,
nA
1.4
Component
Norepineph
10
Dopamine
8
Internal Std
6
Internal Std
4
Norepinephrine
2
7,5
0
Time (min)
10
10,0
20 nM
0
Time (min)
Component
RSD (%)
Conc. (µg/l)
I-cell
300 - 500 nA
Table II. Intra-assay precision of urine sample A and B,
Sample C
Fig. 6. Top: chemical structure of Glucose, Fructose, Lactose and Sucrose.
n= 5 (samples) x 2 (duplicate
injections).
This method
is particularly attractive for the analysis of simple
Norepinephrine
3.8 overlay of 23 48.7
Bottom:
chromatograms of a standard mixture of 2 µM Glusugars in a wide range of food products such as beverages, fruit
cose,
µM Fructose and 4 µM Sucrose in water (20 µl
Epinephrine
6.2 2 µM Lactose, 4 5.1
juices,
and(µg/l)
beer.
injected), ADF 0.01 Hz. The theoretical plate number for the components
Component
RSDmilk
(%) products
Conc.
Dopamine
3.2
225.1
are
N ~ 14.900, 10.900,
12.000 and 17.300 plates/meter, respectively.
Sample A
The performance of the ALEXYS 100 ‘Carbohydrate I’ isocratic
The RSD’s for norepinephrine and dopamine were smaller then
Norepinephrine
2.8
30.6
system is demonstrated using a standard mixture of Glucose,
Linearity
and Detection
Limits
4%. For epinephrine, which
was present
in the sample
in a signifiEpinephrine
1.7 Lactose and16.0
Fructose,
Sucrose in water. In figure 6 an overlay is
cantly lower concentration,
thestandard
RSD wasmixture
slightlythe
higher,
6.2%.
For the
concentration
detection limits (CLOD)
Dopamine
102.5 recorded chromatograms.
shown3.7
of 23 consecutively
were determined to illustrate the sensitivity of PAD detection of
carbohydrates at a gold working electrode. The detection limits
Sample B
C O N C L U S I O N are displayed in Table 2. The CLOD here is based on a 20 µL
Norepinephrine
2.5
20.7
injection
and
defined as the concentration
The ClinRep® complete
kit for
catecholamines
in urine pro- that gives a signal that
Epinephrine
14
3.6
is three times the peak-to-peak noise.
vides a standardised method for the sample preparation of
Dopamine
2.3
115.2
Epinephrine
0
12,5
Number of injections (n)
20 µL
urine samples (A and B). The urine samples were worked-up 5
Temperature
30°C, column and flow cell
Table III. Inter-assay precision (urine sample C,
times and duplicate analysis were performed to determine the
E-cell
E1, E2, E3: 0.05, 0.75, -0.80 Volt n= 4 (samples) x 2 (duplicate injections)
0
5
x 2 (days).
relative standard deviation (RSD, %).
ts, t1, t2, t3: 0.06, 0.5, 0.13, 0.12 seconds
1.6
1.4
0.0
Fig. 1 Analysis of 20 μL ClinCal® urine calibrator. Concetration of
catecholamines in the calibrator sample: 123 μg/L NA, 29.5 μg/L
A and 227 μg/L Dopamine.
nA
Epinephrine
1.8
OH
ALEXYS 100 ‘Carbohydrate I’ isocratic system,
cholamines in the concentration range from 1 – 1000 μg/L [6].
p/n 180.0052
Norepinephrine
Epinephrine
Epinephrine
0.5
Dopamine
nA
1.0
OH H
Sucrose
Page : 3/4
Urine samples were collected
healthy
volun10 nA
Samplefrom an apparently
2 µmol/L
Glucose,
Fructose, Lactose and
To determine the inter-assay precision an urine sample (C) was
Sucrose
in water
teer. and analysed multiple times to determine
the recoveries,
Mobile
phase of the 30
mM NaOH and 1 mM NaOAc, the
mobile 4 times and analysed (duplicate injection), this proceworked-up
LOD, intra- and inter-assay
precision
method.
phase is continuous sparged with dure
Helium
5.0 repeated the next day and the relative standard deviawas
Flow rate
2 mL/min
tion calculated.
The intra-assay precision of the method was determined using two
Vinjection
IS
1.5
H
OH
Lactose
noise. The method is linear for the determination of the cate-
Analysis of urine samples
HPLC
Analysis of ClinChek® controls
For quality control of the analytical determination Recipe
ClinChek® urine controls have been used in both the normal
(level I) and the pathological range (level II).
HO O
H HO
H
Tabel 1
2.0
H
HO
H
H
H
HO
• H2O
O
H
OH
O OH
H
OH
OH
LC-EC Conditions
2.5
H
H
OH
OH
HO
is evident from the obtained RSD values of < 0.4% and < 3% for
the retention times and peak areas of the sugars, respectively.
HO
H
HO
OH
OH H
OH
Glucose
Mixtures of simple sugars, such as mono- and disaccharides can
be determined using PAD under isocratic conditions with high
sensitivity and good reproducibility.
Page : 2/4
H HO
H
H
H
O
O OH
H
OH
HO
Part I - Isocratic analysis of sugars
Page : 2/4
With the ALEXYS 100 ‘’Carbohydrate I’ system a Limit of Detection of 0.2 pmol (on column) for glucose and Lactose could be
reached under the specified conditions. In Fig. 7 an example
chromatogram of a 20 nM sugar mixture is shown to demonstrate
the sensitivity of the method. The analysis of these carbohydrates
at the 100 pmol range can be achieved routinely.
HO
HO
IN URINE
Page : 4/10
Column regeneration
214_001 #01
220_002 #03
Retention time (min)
crises. Nevertheless, deter-
Page
ION
Carbohydrates
not only proTHE• Carboh
vide the most
BRIG
ydrates in food
ENVIRON HTEST
easily access
industry and
•
MEN s LC-EC APPL
ifields in life
EVER Analyte
ble energy
TAL of simpleICAT
sciences
PLOTTED
ANALYSI
source for
monoIONS or
our
FORdisaccharides
• Analyte
S
Bisphenol
body, they
sOUT
of oligosaccharid
A
INTR
also play
es (Maltod
• Analytes
ODU
Catechin
Today,
LC-EC
has been established as a fast
reliable methodan imporextrin)
tantand
s
of polysaccharide
role
CTIO
in many physio
Chloro
N
s (starch
for
the determination
metabolites in plasFlavonoi
phenols
• Analytes
logi, cellulos
ds and phenols of catecholamines and
cal proces
of glycoproteins
nols are e)
and nitr
ses. Some
ma
and urine [1-5].
Phenols
examopheused
ples in
Chloroof urine
in indu
theseofproces
The
ClinRep® complete kit for catecholamines
Recipe
agricul
Antioxid
and nitro
• Chl
stry and
ants
ses are
ture for
orophe
phenols
interce
GmbH (Munich, Germany) is a standardised
kit for
the sample
Due
2,4-dto
several
llular
poses.
nols in
the poor detectio
initro
recognition,
• Nitr
phen
purIn the
industry
n limits of
preparation and routine analysis of urinaryinfectio
catecholamines. In this
ophe-n
ferred
phenoldetectiool (DNP
RI, EC
end they
Polyphenols
fouhas
)
n processes,
and agr
n method
(P)
become the
• 11 EPA ols in indu
in trace analysis nd in
application note the results are reported from analysing urine and
Chroma
iculture
rive
pre- may be
4-nitroph tograph
stry and
certain types
Resverat
phenols
wat. Anion-E r or
..
enol (4-NPy at high pH
samplesrol
using a ClinRep® kit with an ALEXYS 110
LC-EC
sysof cancer
agricul
amperometric
in combinationer. The xchangdrin
e king
2-me
in wat
.
)
The
thylture
detectio
MA
Epicatechin
with
determination
er
4,6-dinitro
n (PAD) is
admissi pulsedC (maxim
tem.
analytical
nowada
2-ch
of carbohyphen
ys theble
lorop method
um
Quercetin
availab
ol (MDN
most
consensitiv
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drates is of
In
for carbohy
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the
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2-nitthis
drate
interest to
EEC
tion
P)note
roph
analysi
Method
ion)
other
countri
we present
the
s [1,2].
polyphenols
solutionenol
in
food industr
a simple
in
2,4-dimet for(2-NP
es for
the determi
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and sensitiv
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nation of sugars drinking
phenols
food hylpheno
d
fields
One Recipe ClinRep® complete kit contains
all the neces-sary
water e system
and oligosa
s based
4-chloro-product
in
l
life
the
(DMP
scienc
on
is
Carbohydrates
ccharid0.5
the ALEXY
)
The dete
es.
3-methylp
es in
µg/l. In
chemicals and (calibration) materials for sample preparation
and
S 10070’s
the US
'Carboh
2,4-d
henol
ction
Metho
Analyte of
ydrate'
ichloroph
protect
envsystem
d
ships
Iodide
(CMP
potentia
ironmen
interes
analysis of 100 assays, excluding the analyticalscolumn.
Urine
s.
)
for 9
enol (DCP
ion age
t include
2,4,6-trich
l
is
phe
tal
optimis
some
ncy (EP
simple
)
Vitamins
made
Anion loropheno
samples
processed using the following
sample
preparation
monoA, C,are
phenols nols. Due
ed
D, E,
a list
or disacchapentachl Exchan
A)
and K
ge
l (TCP
Chromatograph
mV vs.
, especia to the poo by constru
of the
Q10procedure prior to analysis:
rides (such
Ketones > 12)orop
cting
y imp
henol drates)
Ag/AgCl
carbohy
elev
- Under
r dete
ortant
lly 2,4
as glucose
I/E rela
alkaline
(PCP) can be
current
ction
DNP,
is use
phe conditioen mos
and
ubiquinols
Exchange
tioncha
is
sucrose), oligosa
ed ts
a wor
nan
Chromatograph separat
by means nol con ns (pH t
Working approximat d for furth
king pote racteristics
as prio of Anion-tamiy. Carbohydrates
ccharid
pKa values
er exp
• 3 mL acidified urine sample (10 mL conc. 32% HCl per
liter es
ely 200
of
ntial of
ranging betwee
eriments
The
(Maltodextrin),
phenol electrode
are rity acids
120
n 12 and 14.standar weakpoll
utan
either comple
contam nA.
urine) or urine calibrator is mixed with 10 mL
stabilising
re. The
concen
polysa
with
ts.
cchately or partially
backgro 0
ination
low ppb
EPA
trations
based At highdpH
rides
they
value. Due
willhod
ionised
(starch
met
was only
(IS)
and subsequently
agent S and 30 μL internal standard
und
be
, cellulose) addepend
on ing
to the
tion of range or lowe . The con
a con
oncen
problem
and
their pKa is
anion-exchang extreme alkalineliqu
the
glycop
justed to a pH 3.0 – 7.0 using 0.5M
NaOH.
r. At the centrations
conditio
trating
id-liquid
roteins.
atic at
One clea working
e columns
ns only polyme
in sam
high
are
The retentio
extracti ric
ning proc electrode ppb level no
• The mixture is applied to a ClinRep®
sample preparation colples are
lowedfor carbohy
Numer
n time of carbohy suitable
mai
ous analyti
on
cou
ntai
sign
drate
by der
edu
folin the
ld
pKa value
n repr
cal techdrates is inverse
ificant
umns to trap the catecholamines present in the sample.
ivatisat separation.
and increas
oducible re at the end be measure
contam
niques for
es significaGC analysi ly correlat
ion
elution order
d with
ed and
with
of eac
working
ntly with molecu
• The column is subsequently washed with
15carboh
mL HPLC-grade
ydrate anals withlar
in one inaof carbohy
Table
h
day
cap
con
drates
ysis have been
weight.
elec
1
day.
is usually as
ditions.
is suff
tureanion-e
on such
tron The
water to remove interfering compo-nents.
detectio
follows: sugar
icient
developed
LC-E
xchange column
C cond
to
alcohol
di-, tri-, and
In this
n.
over
s elute
s
• 6 mL of eluting reagent E is then
used
elute the catecholathetoyears
itions
higher oligosa
appfirst,
followed by
[6]. The polar,
lication
ccharid
es.
met
mines from the extraction column.
non-volatile,
a HPL mono-,
Colu
hod for
mn
water-soluble
C
the ana
• The eluate is collected, mixed carboh
and 20 μL injected in the
LC
11 EPA
Flow
lysis
rate
ydrates are
Spheriso
phenols
of the
system.
not easily
Mobile
rb ODS
describ
in wat
re-solved.
1.5 ml/m
phase
2, 100x
er is
ed
Also detecti
Sample
4.6 mm,
in
on of
ical dete using elec
50 mM
3 µm
these non-ch
trochem
HAc/NaA
Tempera
ction.
romophoric
100 c, pH
ture
Detecti
are betw
1000
compounds
on limi
nM phen 4.0, 35%
een 25
is difficult.
30
acetonitr
°C
ts
ols, 20
Carexcept
and 220
ile
bohydrates
µl injec
for DNP
ng/l,
hardly have
tion
(0.95
(0.9 µg/l
any
UV absorp
µg/l) and
), TCP
tion and therefo
PCP (6
re
either refract
µg/ml).
ive index (RI)
or
electrochemic
al detection
(EC) is used
for analysis.
Peak areas (μA.s)
(A) and dopamine (DA) are
Glucose
nephrine (NA), epinephrine
Fructose
The catecholamines
norepicolumn
column
silica-based
C18 silica-based
using aa C18
using
Sucrose
INTRODUCTION
C2a
C2a
E
A P PN V I R O
N
LIC
AT I M E N T
ON
A
N OT L
E
Lactose
separation
better separation
Much better
• Much
Page : 1/10
logically increased values,
tec.com
Gentamicine
myan
m
m - www.
even after a hypertensive
myantec.co
myantec.co
Catecholamines
- www.
- info@
c.com
Netherlands sa@myante
crisis.
- www.myantec.com
e - The
- info.u
- [email protected]
Zoeterwoud 21076 - USA
.com
- The Netherlands
NV ntec.com - www.myantec
Carbohydrates
ver MD
- 2382 NV - Zoeterwoude
12 - 2382
- info.usa@mya
Leyden
B.V. - Industrieweg 12 - 2382 NV - Zoeterwoude - The Netherlands - [email protected] - www.myantec.com
- USA
- Industrieweg 12
- Hano
MD 21076 Antec
trieweg
Drive
Antec Leyden B.V.
Drive - Hanover
- Indus
Connelley
Antec (USA) - Suite 110 - 7502 Connelley Drive - Hanover MD 21076Antec
- USA
- [email protected] - www.myantec.com
n B.V.
110 - 7502 Connelley
Leyden
- 7502
Antec (USA) - Suite
B.V. - Industrie
)
Antec (USA)
weg 12 2382 NV
- Suite 110
Antec (USA
- Zoeterw
- 7502 Connelle
oude - The
y Drive Netherlands
Hanover MD
- info@my
21076 - USA
antec.com
- www.my
- info.usa@
Antec
antec.com
myantec.com
Leyden
Phe
- www.my
B.V.
Antec
antec.com nols
(USA
) - Suite Industrieweg
12 110
- 7502
2382
NV Connelle
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y Driv
e ude
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ds 6 - USA
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FOOD
A P P L I C & B E V E R AG E
AT I O N
N OT E
Peak area (μA.s)
side antibiotics
THE SOUNDEST
LC-ECoside
APPLICATIONS
FOR
• Aminoglyco
CLINICAL & DIAGNOSTICS ANALYSIS C2
and
C1a, C2 and
C1, C1a,
e C1,
EVER BUILD
Gentamicine
• Gentamicin
synthesised in the chromaffin
living
ission in
process
Catecholamines
by a fermentation
cells of the adrenal medulla
rotransm
of the
(NA),
es
pinal fluid
Aminoglycosid
is
renaline s are Serotonin
constituent
main
and the
norad
and the sympathetic nervous
Cerebros
of
s animals Amikacin
s 5- and
analysis
consciou
Method
Method
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column.
a C18 silica-based
the interval between two
ne
Noradrenali
36
Phenol derivatives
11
12*
10
220_002 #03
Page : 1/4
Dopamine
®
®
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221-019
Lactose
CATIO
4
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Page : 1/4
EVER
6x10
221-007
www.myantec.com
1 #03
INE
DOPAM
CLINREP® KIT FOR THE ANALYSIS OFANALY
ALINE,GENTAMICIN SULPHATE
IONS
SIS OF CAR
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BOHYDRAT
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THE SMAR NCE ANAL
NEUROSCIE ERMINDED
MAST
Urine
pharmaceutical dosage forms
FOOD & BEVERAGE
Toll free (888) 572 0012
214_001 #01
Schematics ALEXYS
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INTRODUCTI
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Spectinomycin
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221-020
Magic Diamond™ - Unprecedented
Performance in HPLC-EC
2
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Proteins
Glutathioneandother(di-)sulfides
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Page :
217_013 #05
6
USA
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221-010
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Industrieweg 12
2382 NV Zoeterwoude
Schematics ROXY™ EC/LC System for on-line EC/LC/MS,
with ReactorCell as part of the fluidic pathway of the
autosampler1) for automated activation
(oxidation/reduction), adduct formation and sample
screening. (patent pending).
8
221-003
2
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The Netherlands
Schematics
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Tel. +31 (0)71 581 33 33
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Fax +31 (0)71 581 33 34
metus, ac tincidunt erat. Nulla adipiscing, massa ut rutrum sed mollis tincidunt, nisl sem posuere leo, non
[email protected]
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volutpat lorem. Donec luctus fermentum risus at congue.
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ornare.
Application folders
9
PHARMACEUTICAL
& BIOTECH
6
2x10
Pellentesque volutpat aliquam erat in suscipit.
Morbi in tempor ligula.
4
5
Pumps
tibulum sed tempus magna. Nulla ultricies dui at
2
tetrazepam
Simulation of the4x10
oxidative metabolism
of caspase
binding of Aminoglycoside Antobiotics
221-005
The analysis
EC
radio-ligands (CBRs) by online electrochemistry-HPLC-MS
ullamcorper lacus sapien sit amet lectus. Vesnulla consequat sit amet sagittis nunc consequat.
Aminoglycosiden
3
1
0
CLINICAL & DIAGNOSTICS0
Phasellus in lacus arcu. Integer imperdiet, ipsum
Peptides
Three min time resolution in on-line microdialysis
Urine
5
5
intensity [cps]
Lorem ipsum dolor sit amet, consectetur adipisc-
Phase II: adduct formation, e.g.,
conjugation of the oxidized compound with
GSH, HSA, drug-protein binding studies, etc.
Drugs
-
hplc/ec
ing elit. Phasellus lacinia bibendum scelerisque.
Schematics ROXY™ EC System for on-line EC/MS:
Phase I: oxidation reactions, e.g.,
oxidative metabolism, biotransformation reaction, etc.
ecD - HPlc/ec PoSterS
-
Any Massaspectrometer
Clarity software
electrocHemiStry anD liqUiD cHromatograPHy
US!
-
Clarity software
HPlc/ec
NEUROSCIENCE
Decade II
ROXY™ EC/LC System
ecD
focUS on
ec ecD HPlc/ec tecHnology
PHARMACEUTICAL & BIOTECH
-
PWA - 489
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ec
®
MPX-544
Clarity software
Ec/Lc/Ms
Focus on
Ec/Ms Ec/Lc/Ms TEchnoLogy
Ec/Lc/Ms posTErs
Any Massaspectrometer
Ec/Ms
ELEcTrochEMisTry and Mass spEcTroMETry
FEEL FrEE To conTacT us!
Pump
Pumps
Ec
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. Box 111 32911 tion of
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Documentation
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Invitation to Antec’s support site
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F O O D & B E V E R AG E
220_002 #03
Page : 1/10
ANION-EXCHANGE CHROMATOGRAPHY AND
PULSED AMPEROMETRIC DETECTION
Customer satisfaction is and has always been our first
priority. To support our customers we have created a
support website which is now at your disposal.
THE FINEST LC-EC APPLICATIONS FOR
FOOD & BEVERAGE ANALYSIS
EVER PROCESSED
INTRODUCTION
Carbohydrates not only provide the most easily accessible energy source for our
body, they also play an impor-
Our knowledgebase contains all the relevant tips and tricks
Bisphenol A
tant role in many physiologi-
Catechins
cal processes. Some exam-
from years of experience in LC-ECD.
Antioxidants
It offers a 24/7 support, always available when needed.
Flavonoids and phenols
ples of these processes are
Phenols
intercellular recognition,
Polyphenols
infection processes, and
certain types of cancer. The
Resveratrol
determination of carbohy-
Epicatechin
drates is of interest to the
Quercetin
food industry and many fields
other polyphenols
Carbohydrates
Iodide
•
•
•
•
•
Carbohydrates in food industry and fields in life sciences
Analytes of simple mono- or disaccharides
Analytes of oligosaccharides (Maltodextrin)
Analytes of polysaccharides (starch, cellulose)
Analytes of glycoproteins
Due to the poor detection limits of RI, EC has become the preferred detection method in trace analysis. Anion-Exchange
Chromatography at high pH in combination with pulsed
amperometric detection (PAD) is nowadays the most sensitive
analytical method available for carbohydrate analysis [1,2].
In this application note we present a simple and sensitive system
solution for the determination of sugars and oligosaccharides in
food products based on the ALEXYS 100 'Carbohydrate' systems.
in life sciences.
Method
Analytes of interest include
Anion Exchange Chromatography - Under alkaline conditions (pH
> 12) carbohydrates can be separated by means of AnionExchange Chromatography. Carbohydrates are weak acids with
pKa values ranging between 12 and 14. At high pH they will be
either completely or partially ionised depending on their pKa
value. Due to the extreme alkaline conditions only polymeric
anion-exchange columns are suitable for carbohydrate separation.
The retention time of carbohydrates is inversely correlated with
pKa value and increases significantly with molecular weight. The
elution order of carbohydrates on such anion-exchange columns
is usually as follows: sugar alcohols elute first, followed by mono-,
di-, tri-, and higher oligosaccharides.
simple mono- or disaccha-
rides (such as glucose and
Q10
sucrose), oligosaccharides
ubiquinols
(Maltodextrin), polysaccharides (starch, cellulose) and
glycoproteins.
Numerous analytical techniques for carbohydrate analysis have been developed
over the years [6]. The polar,
non-volatile, water-soluble
carbohydrates are not easily
re-solved. Also detection of
these non-chromophoric
compounds is difficult. Carbohydrates hardly have any
UV absorption and therefore
either refractive index (RI) or
electrochemical detection
(EC) is used for analysis.
Carbohydrates
Register today at www.myantec.com
go to support and have free access !
®
37
Distributors
Alexys 110 REPRESENTATION
WORLDWIDE
Standardised for routine analysis
ARGENTINA
Analitica SA
Jorge Newbery 1840 of.3
Buenos Aires
Phone: +54 (11) 4772 4922/4450
Fax: +54 (11) 4775 7549
E-mail: [email protected]
Site: www.analiticasa.com.ar
AUSTRALIA
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Rydalmere NSW 2116
Phone: +61 (2) 96844200
Fax: +61 (2) 96844055
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AUSTRIA
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Phone: +43 (1) 982 7575
Fax: +43 (1) 982 7589
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Beijing 100176
Phone: + 86 (10) 5402 9811
Fax: + 86 (10) 5102 9820
Site:
http://huize.instrument.com.cn
COLOMBIA
CiCs Instruments S.A. (Crelab
Instruments Colombo Sueca)
Calle 147 No. 7B-64
Bogota - Colombia
Phone: +57 (1) 615 23 63
Fax: +57 (1) 608 03 59
CZECH REPUBLIC
Science Instruments and Software
s.r.o.
Fetrovská 59
160 00 Praha 6
Phone: +420 (246) 037 483
Fax: +420 (246) 030 500
Site: www.sisw.cz
DENMARK
Biolab A/S
Sindalsvej 29
DK-8240 Risskov
Phone: +45 (86) 212866
Fax: +45 (86) 212301
Site: www.biolab.dk
EGYPT
Arab Consulting Engineers-Osman
A. Azzam & Co.
Building No.5
Street No.77, Al Maadi
Cairo 11728
Phone:+20 (2) 750 8724, +20 (2)
378 1577
Fax: +20 (2) 378 3845
E-mail:
[email protected]
FINLAND,ESTLAND
Ordior Oy
Konalantie 47A
00390 Helsinki
Phone: +358 (9) 530 8000
Fax: +358 (9) 530 80010
Site: www.ordior.fi
FRANCE
Alpha M.O.S.
20 Av. Didier Daurat
31400 Toulouse
Phone: +33 (5) 6247 6459
Fax: +33 (5) 6154 5615
Site: www.alpha-mos.com
GERMANY
Axel Semrau GmbH & Co.KG
Stephansbecke 42
45549 Sprockhövel
Phone: +49 (2339) 12090
Fax: +49 (2339) 6030
Site: www.axelsemrau.de
GREECE
Biosolutions
Ltd249 Mesogion
Ave.154 51 N. Psychiko, Athens
Phone: +30 210 6753453
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Web site: www.biosolutions.gr
HUNGARY, SERBIA &
MONTENEGRO
ABL&E - JASCO Hungary Ltd.
Fehérvári út 130
H-1116 Budapest
Hungary
Phone: +36 (1) 209 3538
Fax: +36 (1) 279 0472
Site: www.ablelab.com
INDIA
Spinco Biotech Pvt Ltd
Post Box No. 6114
No.4 Vaidyaram Street
T. Nagar, Chennai 600 017
India
Phone : +91 (44) 2434 0174
Fax : +91 (44) 2434 0761
Site: www.spincotech.com
INDONESIA
P.T. Besha Analitika
Komplek Ruko Pulogadung Trade
Centre Blok 8B No. 59
Kawasan Industri Pulogadung
Jl. Raya Bekasi KM 21
Jakarta Timur
Phone: +62 (21) 4683 5883
Fax: +62 (21) 4683 2968
IRAN
Shimi Tajzieh Sina Engineering Co,
Ltd.
No. 19, Amaj St. 33rd Asadabadi St.
Teheran 14346
I.R. Iran
Phone: +98 (21) 88702730
Fax: +98 (21) 88722029
Site: www.shimisina.com
IRELAND
APEX Scientific Ltd.
Unit F12,
Maynooth Business
Campus, Maynooth,
Co Kildare
Phone: +353 (1) 6854 686
Fax: +353 (1) 5054 665
Site: www.apexscientific.ie
ISRAEL
Lab Experts
P.O.B. 3383
Bat Yam 59134
Phone: +972 (3) 657 3785
Fax: +972 (3) 657 3785
Site: www.lab-experts.com
ITALY
Alfatech S.r.l.
Via Scarsellini 97
16149 Genova
Phone: +39 (010) 469 9369
FAX: +39 (010) 469 9377
Site: www.alfatechspa.com
JAPAN
Sanwa Tsusho Co., Ltd.
13-2, Nishi-Gotanda 3-chrome
Shinagawa-ku
Tokyo 141-0031
Phone: +81 (3) 3492 6300
Fax: +81 (3) 3492 6311
Site: www.sanwatsusho.com
JORDAN
Hijaz Electronic & Scientific Supplies Est.
P.O. Box 925133
Amman 11190 Jordan
Phone: +962 (6) 5562195
Fax: +962 (6) 5562194
Distributors
MALAYSIA
Research Instruments Sdn. Bhd.
Jalan SS25/2, Taman Bukit Emas,
47301 Petaling Jaya,
Selangor
Phone: (603) 7804 8600
Fax: (603) 7804 8599
Site: www.ri.com.my
MEXICO
EVOELUTION SA de CV
Miguel N. Lira 292
Col. Villa de Cortés
03530 Mexico, D.F.
Phone: +52 (55) 5579 0759
Fax: +52 (55) 5579 0742
E-mail:
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Site: www.evoelution.com
MOROCCO
SCOMEDICA
N°22 Résidence Abir Rue
Mausolée
Quartier de Hôpitaux
20100 Casablanca
Phone: +212 (22) 864578 /
+212 (22) 864608
Fax: +212 (22) 864579
NETHERLANDS
Antec Leyden B.V.
Industrieweg 12
2382 NV Zoeterwoude
Phone: +31 (71) 5813333
Fax: +31 (71) 5813334
NEW ZEALAND
Shimadzu Scientific Instruments
(Oceania) Pty. Ltd.
Unit A, 13-15 Collard Place
Henderson
Auckland
New Zealand 1230
Phone: +64 (9) 8367750
Fax: +64 (9) 8367757
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NORWAY
Bergman AS
Slynga 2
N-2005 Raelingen
Phone: +47 (63) 835 600
Fax: +47 (63) 835 610
Site: www.bergman.no
PAKISTAN
Analytical Measuring Systems
(Pvt.) Ltd.
AMS House
Plot # 14C,Main Sehar Commercial
Avenue
Commercial Lane 4
Khayaban-e-Sehar
D.H.A. Phase 7
Karachi
Phone: +92 (21) 5345581, 5340747
Fax: +92 (21) 5345582
Site:
www.amspvtltd.com/AMS
PERU
Electromedica Peruana S.A.
Analytical Division
Av. Prolong
J. Prado Oeste 630
Mag. Del Mar, Lima
Phone: +51 (1) 460 1317
Fax: +51 (1) 460 1448
E-mail:
[email protected]
POLAND
SHIM-POL A.M. Borzymowski
Reszka Spolka Jawna
Ul. Lubomirskiego 5
05-080 Izabelin k. Warszawy
Phone: +48 (22) 722 7048/49/50
Fax: +48 (22) 722 7051
Site: www.shim-pol.pl
PORTUGAL
Biocitek S.A. - Grupo Taper Portugal
Departamento de Equipamentos
Analiticos
Est. da Barrosa, Elospark 3
2725-193 Mem-Martins, Sintra
Phone: +351 (21) 922 7200
Fax: +351 (21) 922 8800
ROMANIA
S.C. VIOLA total S.R.L.Str.
Telescopului, Nr. 13A014368
Sector 1
Bucharest
Phone: +40 (21) 233 2339
Fax: +40 (21) 233 2340
[email protected]
Site: www.viola.ro
RUSSIA
ANALIT Co. Ltd.
Bolshoi Prospect V.O.
Building 31, Office 108
199004 St.Petersburg
Phone: +7 (812) 325 5502
Fax: +7 (812) 320 6917
Site: www.analit-spb.ru
SAUDI ARABIA
K.I. Abdulkadir & Partners Co. Ltd.
P.O. Box 9547
Jeddah 21423
Saudi Arabia
Phone: +966(2)6405846
Fax: +966(2)6404710
Site: www.kiaksa.com
SINGAPORE
Research Instruments PTE Ltd
26 Ayer Rajah Crescent,#03-10
Singapore 139944
Phone:+65 6775 7284
Fax: +65 6775 9228
Site: www.ri.com.sg
SLOVAKIA
Shimadzu Slovakia
Ul. Dr. V. Clementisa 10
82102 Bratislava
Slovakia
Phone: +421 (2) 48200081-4
Fax: +421 (2) 48200085
Site: www.shimadzu.sk
SPAIN
Ingenieria Analitica s.l.
Avda.de Cerdanyola, 73
Apartado 282
08190 Sant Cugat Del Vallès
Barcelona
Phone: +34 (93) 590 2850
Fax: +34 (93) 675 0516
Site:
www.ingenieria-analitica.com
SWEDEN
Crelab Instruments AB
Kalvhagen 20
SE-42750 Billdal
Phone: +46 (31) 748 9626
Fax: +46 (31) 748 9426
Site: www.crelab.se
SYRIA
NM-ANALYSIS
Contact person:
Mr. Hussam Nabhani
Damascus
Mobile Phone: + 963 944396912
Fax: + 963 (11) 44691167
TAIWAN
Anatech Co., Ltd.
2F, No. 245, Nung An St.
Taipei 104
Phone: +886 (2) 2500 6520
Fax: +886 (2) 2502 8195
THAILAND
Sithiporn Ass. Co., Ltd
451 Sirinthorn Rd.
Bangbumru, Bangplud
Bangkok 10700
Phone: +66 (2) 8819244
Fax: +66 (2) 433 1679
Site: www.sithiporn.co.th
TUNISIA, ALGERIA
Science & Technology (Satco)
54, Av. Tahar Ben Achour
1082 Mutuelleville Tunis
Phone: +216 (1) 794 312 / 788 690
Fax: +216 (1) 792 354
TURKEY
Dolunay Teknik Cilhazlar Ltd. Sti.
Ornek Tepe Cad No. 165/1
34445 Sutluce Istanbul
Phone: +90 (212) 210 5435
Fax: +90 (212) 210 5380
Site: www.dolunay.com
U.A.E., QATAR, YEMEN,
KUWAIT, BAHRAIN
EMPHOR FZCO
P.O. Box 61232
Jebel Ali Free Zone
Dubai
Phone: +971 (4) 8830233
Fax: +971 (4) 8830133
Site: www.emphor.biz
USA
Antec (USA)
P.O. Box 111418
Palm Bay, FL 32911
USA
Toll free (888) 572 0012
Phone: (321) 473-4202
Fax: (321) 821-1938
KOREA
Young Lin Instrument Co. Ltd.
899-6 Hokye 2-dong,
Dongan-gu, Anyang-si
Kyounggi-do 431-836
Tel:+82 (31) 428 8700
Fax:+82 (31) 428 8787
Site: www.younglin.com
®
39
®
Antec (Leyden) B.V.
Antec (USA)
Industrieweg 12
P.O. Box 111418
2382 NV Zoeterwoude
Palm Bay, FL 32911
The Netherlands
USA
Tel.+31 (0)71 581 33 33
Toll free(888) 572 0012
Fax+31 (0)71 581 33 34
Phone (321) 473-4202
[email protected]
Fax
www.myantec.com
[email protected]
Distributor
Distributed by ARC Sciences Ltd., PO Box275, Alton, Hampshire, GU4 9FJ – t:01420 549922
[email protected]
www.arcsciences.com
(321) 821-1938
15200910
www.myantec.com

Alexys Electrochemical System Solutions

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