Carbohydrate Analysis using HPLC
with PAD, FLD, Charged Aerosol Detection,
and MS Detectors
Bruce Bailey, Paul Ullucci, Rainer Bauder, Marc Plante, Chris Crafts, Ian Acworth
Thermo Fisher Scientific, 22 Alpha Road, Chelmsford, MA, USA
Overview
Purpose: Sensitive applications for the analysis of simple carbohydrates, either as
mono- or disaccharides, or as fairly small glycans liberated from glycoproteins typically
containing 6–11 residues, are desired to enhance sampling rates. Simple, rapid and
accurate methods have been developed for the analysis of carbohydrates using
various chromatography and detection solutions.
Methods: A sensitive pulsed amperometric detection method for the analysis of simple
carbohydrates was developed and examples are shown for impurity analysis of sugars
used in positron emission tomography (PET). A second chemistry for the analysis of
fluorescent derivatives of glycans from glycoproteins is described. The direct analysis
of simple sugars using HILIC-mode chromatography with charged aerosol detection is
described. Examples showing the use of this technique for their direct determination in
fruit juices are shown. Finally, orthogonal use of charged aerosol detection and MS
detectors for the analysis of glycans from glycoproteins is described.
Results: These methods enable the rapid separation of carbohydrate compounds at
low levels and with minimal matrix interference.
Introduction
Carbohydrates are difficult to analyze because they have similar physical and chemical
characteristics, and do not have a suitable chromophore for UV detection. Several
different HPLC methods using various detector strategies (pulsed amperometric
electrochemical, fluorescence following derivatization, charged aerosol detection and
mass spectrometry) were developed to help study carbohydrates and examples for
each approach will be presented. Although fluorescent tags improve the
chromatographic resolution and detector sensitivity, they can lead to increased assay
variability. Different HPLC modes can be used for carbohydrate separations, with ion
exchange, hydrophilic interaction liquid chromatography (HILIC) and reversed phase
(RP) on porous graphite column (PGC) being the most common. HPLC enables the
development of simpler chromatographic methods with direct detection using PAD
or mass detectors such as ELSD, MS, and charged aerosol detection. The Thermo
Scientific Dionex Corona Charged Aerosol Detector (CAD™) is an ideal detector when
combined with HILIC or RP/PGC for measuring different carbohydrates. It is a masssensitive detector that can measure any non-volatile, and many semi-volatile
compounds, typically with low nanogram sensitivity. Unlike ELSD, it shows high
sensitivity, wide dynamic range, high precision, and more consistent inter-analyte
response, independent of chemical structure. For the analysis of glycans liberated from
glycoproteins, the utility of the LC-MS with charged aerosol detection platform is
illustrated where the Corona CAD is used for quantitative analyses while the MS
provides structural verification. The advantage of this approach over methods using
fluorescent tags is discussed.
Methods
General considerations for Pulsed Amperometric Detection
The Thermo Scientific Dionex UltiMate 3000 with PAD platform consisted of a basecompatible HPLC system, a Thermo Scientific Dionex Coulochem III electrochemical
detector and a gold working electrode. Carbohydrates were separated using ion
exchange and determined under basic conditions using a four-pulse waveform. It is
essential to make sure no titanium is in the flow path as its degradation under basic
conditions can lead to deterioration of column and electrode performance.
Carbohydrate Analysis by HPLC-EC PAD:
Pump:
Autosampler:
Flow:
Column:
Temperature:
Injection
volume:
Mobile Phase:
EC detector:
EC Parameters:
Range:
Thermo Scientific Dionex ISO-3100 SD
Thermo Scientific Dionex WPS-3000TSL Analytical Autosampler
Isocratic at 0.50 mL/min. with constant He purge
Thermo Scientific CarboPak: PA20, 3 x 150 mm, 6.5 µm
32 °C
50 µL partial loop
50 mM sodium hydroxide (NaOH), prepared from pellets, 99.99%,
semiconductor grade
Coulochem™ III, Thermo Scientific Dionex model 5040 cell with
Au Target: 25 µm Mylar
E1
+200 mV
500 ms
AD 300 ms
E2
-2000 mV
10 ms
E3
+600 mV
10 ms
E4
-100 mV
10 ms
200 nC
2 Carbohydrate Analysis using HPLC with PAD, FLD, Charged Aerosol Detection, and MS Detectors
Analysis of Glycan Derivatives with F
Flow:
1.0 mL/min.
Column:
TSKgel® Amide-80, 4
Temperature:
35 °C
Injection volume: 5 µL partial loop
Mobile Phase A: 50 mM ammonium fo
Mobile Phase B: Acetonitrile
Gradient:
65%B to 53%B in 24
equilibration from 27
Fl Detector:
Ex 320 nm; Em 420
Glycan
N-Glycanase (2 µL)
derivative
then placed overnigh
formation:
carbohydrates were
SPE. Glycans were e
water/0.1% TFA. Sam
4 °C. To dried sampl
heated at 65 °C for 3
labeling reagent was
3 times) and sample
reconstituted with 20
vial for injection.
Direct Carbohydrate Analysis in Fruit
Flow:
Isocratic at 1.4 mL/m
Column:
Shodex™ Asahipak® N
Temperature:
55 °C
Post column tempera
Inj. volume:
2 µL for sample, 5 µL
Mobile Phase:
78% Acetonitrile, 22%
Detector:
Corona™ ultra RS™
Nitrogen: 35 psi Cor
Sample
To 1 gram juice samp
Preparation:
then centrifuge for 2 m
Glycoprotein Analysis by parallel Cha
Flow:
1.0 mL/min.
Column:
Thermo Scientific PG
Temperature:
55 °C
Post column tempera
Inj. volume:
5 µL partial loop
Mobile Phase:
MeCN/H2O/0.1% TFA
Detector:
Corona ultra RS
Nitrogen: 35 psi Cor
Sample
N-linked glycans were
Preparation:
recombinant N-glycan
released either by tra
nonreductive ammon
samples was perform
extraction; collecting
Secondary cleanup o
(collect flow-through)
Results and Discuss
Direct Carbohydrate Analysis with PAD
Carbohydrates are commonly measured
chromatography in combination with puls
PAD method was developed using the Co
pulse mode. The chromatogram shown in
carbohydrate standards (1 ng on-column
not shown) and the assay reproducibility
sensitivity of this method can reach a lim
column (data not shown). This method w
[18F]fluoro-D-glucose (FDG), which is the
positron emission tomography. The synth
displacement and hydrolysis. A contamin
2-chloro-D-glucose (ClDG). Limits for ClD
soon by the FDA. Due to the sensitivity a
procedures are not required. HPAC-PAD
method for determining carbohydrates (F
PA20 anion exchange column using an a
resolution requirements set by the USP m
ydrates, either as
glycoproteins typically
Simple, rapid and
hydrates using
the analysis of simple
rity analysis of sugars
for the analysis of
. The direct analysis
d aerosol detection is
irect determination in
detection and MS
ed.
drate compounds at
physical and chemical
etection. Several
amperometric
erosol detection and
and examples for
ove the
d to increased assay
eparations, with ion
and reversed phase
HPLC enables the
ection using PAD
ction. The Thermo
n ideal detector when
drates. It is a massemi-volatile
it shows high
ent inter-analyte
glycans liberated from
tion platform is
s while the MS
ver methods using
onsisted of a basem III electrochemical
rated using ion
lse waveform. It is
dation under basic
ormance.
ytical Autosampler
ge
m, 6.5 µm
om pellets, 99.99%,
del 5040 cell with
0 ms
Analysis of Glycan Derivatives with Fluorescence Detector
Flow:
1.0 mL/min.
Column:
TSKgel® Amide-80, 4.6 x 150 mm, 3 µm
Temperature:
35 °C
Injection volume: 5 µL partial loop
Mobile Phase A: 50 mM ammonium formate in water pH=4.4
Mobile Phase B: Acetonitrile
Gradient:
65%B to 53%B in 24 min, 0%B from 24.5 min to 26.5 min, reequilibration from 27 min to 35 min.
Fl Detector:
Ex 320 nm; Em 420 nm
Glycan
N-Glycanase (2 µL) was added to 100 µg protein in 50 µL buffer
derivative
then placed overnight at 37 °C. Following release from protein,
formation:
carbohydrates were cleaned up using Thermo Scientific Hypercarb
SPE. Glycans were eluted from SPE using 40% acetonitrile/60%
water/0.1% TFA. Samples were dried in a refrigerated speedvac at
4 °C. To dried samples, 20 µL 2-AA solution was added, mixed and
heated at 65 °C for 3 hours. After reaching room temperature, excess
labeling reagent was removed with 1 mL acetone wash (repeated
3 times) and samples were dried with a speedvac. Samples were then
reconstituted with 200 µL MP A and then transferred to an autosampler
vial for injection.
Direct Carbohydrate Analysis in Fruit Juice with Charged Aerosol Detection
Flow:
Isocratic at 1.4 mL/min.
Column:
Shodex™ Asahipak® NH2P-50 4E 4.6 x 250 mm, 5 µm
Temperature:
55 °C
Post column temperature: 30 ºC
Inj. volume:
2 µL for sample, 5 µL for standards
Mobile Phase:
78% Acetonitrile, 22% Water
Detector:
Corona™ ultra RS™
Nitrogen: 35 psi Corona filter: Corona
Sample
To 1 gram juice sample, add 20 mL of 70% acetonitrile, mix, and
Preparation:
then centrifuge for 2 min @ 13,000 RPM
Glycoprotein Analysis by parallel Charged Aerosol Detection and MS
Flow:
1.0 mL/min.
Column:
Thermo Scientific PGC Hypercarb 4.6 x 150 mm, 5 µm
Temperature:
55 °C
Post column temperature: 30 ºC
Inj. volume:
5 µL partial loop
Mobile Phase:
MeCN/H2O/0.1% TFA: 4%−18% Acetonitrile gradient in 40 min
Detector:
Corona ultra RS
Nitrogen: 35 psi Corona filter: Corona
Sample
N-linked glycans were released by standard procedures using
Preparation:
recombinant N-glycanase or endo-H. O-glycans were chemically
released either by traditional reductive β-elimination (RBE),1 or by
nonreductive ammonia/ammonium carbonate (NAC).2 Cleanup of
samples was performed by cation-exchange SPE (solid-phase
extraction; collecting flow-through) and borate evaporation.
Secondary cleanup of polypeptides using a C18 SPE
(collect flow-through) was performed.
Results and Discussion
Direct Carbohydrate Analysis with PAD
Carbohydrates are commonly measured using high performance anion exchange
chromatography in combination with pulsed amperometric detection (PAD). A simple
PAD method was developed using the Coulochem III electrochemical detector with
pulse mode. The chromatogram shown in Figure 1 illustrates replicate injections of
carbohydrate standards (1 ng on-column). Good retention stability was observed (data
not shown) and the assay reproducibility over 14 hours is shown in Table 1. The
sensitivity of this method can reach a limit-of-detection (LOD) of less than 100 pg oncolumn (data not shown). This method was used for impurity testing of 2-deoxy-2[18F]fluoro-D-glucose (FDG), which is the most widely used radiopharmaceutical for
positron emission tomography. The synthesis of FDG involves nucleophilic
displacement and hydrolysis. A contaminant resulting from both processes is 2-Deoxy2-chloro-D-glucose (ClDG). Limits for ClDG have been set by USP, CMC, and EP and
soon by the FDA. Due to the sensitivity and selectivity of PAD, sample preparation
procedures are not required. HPAC-PAD is a well established, sensitive, and selective
method for determining carbohydrates (Figure 2). The separation uses a CarboPak™
PA20 anion exchange column using an alkaline MP (50 mM NaOH) and meets the
resolution requirements set by the USP method.
3 Carbohydrate Analysis using HPLC with PAD, FLD, Charged Aerosol Detection, and MS Detectors
Table 1. Reproducibility statistics for 14
Glucose
Fruc
Height
Area
Height
Mean
178.3
15.9
85.9
SD
1.61
0.21
0.93
RSD
0.90
1.33
1.08
FIGURE 1. Triplicate injections of carb
using anion exchange chromatograph
20.0
nC
10.0
2
1
3
-5.0
0.00
0.50
1.00
1.50
2.00
FIGURE 2. Chromatogram illustrating
FDG/CLDG / mannose using anion ex
900
nC
750
625
500
375
250
125
-100
0.0
1.0
2.0
3.0
4.0
Glycan Analysis with Fluorescence De
The analysis of glycans using fluorescen
suitable derivative. In this case the highly
derivative was prepared according to the
technical note illustrating a similar fluores
FIGURE 3. Carbohydrate separation o
HPLC with fluorescence detection.
6
5
4
3
2
1
0
7.5
10
12.5
Table 1. Reproducibility statistics for 14 hour run (50 ng standard)
Glucose
2-aminobenzamide (AB) has been publis
proposed structure assignments shown in
analysis.
Lactose
Height
Area
Height
Area
Height
Area
Height
Area
178.3
15.9
85.9
8.7
44.6
5.1
95.2
12.9
SD
1.61
0.21
0.93
0.19
0.51
0.11
0.90
0.21
RSD
0.90
1.33
1.08
2.15
1.15
2.23
0.95
1.59
Analysis of Carbohydrates in Fruit Jui
The analysis of simple carbohydrates wa
in HILIC mode (Figure 4). This column, u
separation while offering good stability an
speed of separation of the various carboh
the water content and elevating the colum
eluted in less than 16 minutes and the LO
5 µL injection), which is much better than
was applied to the measurement of carbo
Other food sugars in honey and different
successfully analyzed. The method can b
in a variety of food products. This techniq
that only possess weak chromophores w
sample preparation
FIGURE 1. Triplicate injections of carbohydrate standards (1 ng on-column)
using anion exchange chromatography and PAD detection.
20.0
nC
10.0
FIGURE 4. Carbohydrate measuremen
aerosol detection.
2
1
3
osol Detection
m, 5 µm
Sucrose
Mean
to 26.5 min, re-
ein in 50 µL buffer
ase from protein,
Scientific Hypercarb
% acetonitrile/60%
gerated speedvac at
as added, mixed and
m temperature, excess
one wash (repeated
vac. Samples were then
ferred to an autosampler
Fructose
-5.0
0.00
25.0
min
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
4.50
5.00
FIGURE 2. Chromatogram illustrating the separation and detection of 10 µg/mL
FDG/CLDG / mannose using anion exchange chromatography and PAD detection.
tonitrile, mix, and
900
20.0
15.0
10.0
nC
750
and MS
pA
5.0
625
0.0
500
m, 5 µm
375
adient in 40 min
ocedures using
s were chemically
ation (RBE),1 or by
NAC).2 Cleanup of
PE (solid-phase
evaporation.
8 SPE
nion exchange
n (PAD). A simple
cal detector with
ate injections of
was observed (data
Table 1. The
ss than 100 pg ong of 2-deoxy-2harmaceutical for
eophilic
ocesses is 2-DeoxyCMC, and EP and
mple preparation
sitive, and selective
uses a CarboPak™
) and meets the
-4.0
0.0
250
125
-100
0.0
2.0
4.0
6.0
8.0
FIGURE 5. Direct analysis of carbohyd
min
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
100
Glycan Analysis with Fluorescence Detection
The analysis of glycans using fluorescence detection requires the formation of a
suitable derivative. In this case the highly fluorescent 2-anthranilic acid (2-AA)
derivative was prepared according to the technique described by Anumula.3 A
technical note illustrating a similar fluorescence method for glycan profiling using
1
2
3
4
5
pA
80
60
FIGURE 3. Carbohydrate separation of 2-AA derivatives using HILIC-mode
HPLC with fluorescence detection.
40
FLD, Ex=320,
Ex=320, Em=420
FLD,
Em=420
20 5
4
3
2
1
66
-5
0.0
55
2.0
4.0
6.0
8.0
44
Glycoprotein Analysis with Charged Ae
33
22
1
1
0
0
The Corona CAD and MS use the same v
techniques can also be used orthogonally
the glycans and quantitative data, while M
is shown in Figure 6 for the analysis of gly
powerful approach to glycan identification
90%
90%
10%
10%
7.5
7.5
10
10
12.5
12.5
15
15min
17.5
17.5
20
20
4 Carbohydrate Analysis using HPLC with PAD, FLD, Charged Aerosol Detection, and MS Detectors
22.5
22.5
25
25
)
Lactose
Height
Area
95.2
12.9
0.90
0.21
0.95
1.59
FIGURE 6. Fetuin: reductive β-elimina
MS QTof (+ mode) using a Hypercarb P
detection and MS.
2-aminobenzamide (AB) has been published by Thermo Fisher Scientific.4 The
proposed structure assignments shown in Figure 3 were based on accurate mass
analysis.
Analysis of Carbohydrates in Fruit Juices using Charged Aerosol Detection
The analysis of simple carbohydrates was performed using a polymeric amino column
in HILIC mode (Figure 4). This column, unlike silica-based columns, provides adequate
separation while offering good stability and low column bleed. Improved selectivity and
speed of separation of the various carbohydrate species was achieved by decreasing
the water content and elevating the column temperature. Simple carbohydrates were
eluted in less than 16 minutes and the LOD was ~5 ng on-column (1 µg/mL using a
5 µL injection), which is much better than those achieved by RI or ELSD. This method
was applied to the measurement of carbohydrates in fruit juice samples (Figure 5).
Other food sugars in honey and different corn syrups (data not shown) have also been
successfully analyzed. The method can be used to easily determine sugar abundance
in a variety of food products. This technique is a good example of detecting analytes
that only possess weak chromophores with very simple analytical conditions and
sample preparation
ng on-column)
4.50
5.00
ection of 10 µg/mL
y and PAD detection.
Gly
pA
0
20.0
15.0
5.0
0.0
-4.0
0.0
min
2.0
4.0
6.0
8.0
10.0
12.0
14.0
16.0
18.0
20.0
FIGURE 5. Direct analysis of carbohydrates in various fruit juice samples.
min
9.0
10.0
100
formation of a
acid (2-AA)
numula.3 A
profiling using
1
2
3
4
5
OJ
Apple
Cranberry
Grape
200ug/ml
CAD_1
CAD_1
CAD_1
CAD_1
CAD_1
pA
80
40
FLD, Ex=320, Em=420
20 5
4
3
2
1
-5
0.0
min
2.0
4.0
6.0
8.0
10.0
12.0
14.0
16.0
18.0
20.0
Glycoprotein Analysis with Charged Aerosol and MS detection
The Corona CAD and MS use the same volatile mobile phase. The two analytical
techniques can also be used orthogonally with the Corona CAD providing the profile of
the glycans and quantitative data, while MS is able to provide structural information. This
is shown in Figure 6 for the analysis of glycans from fetuin. The combined technique is a
powerful approach to glycan identification and quantification.
22.5
10.00
12.00
14.00
16.00
18.00
20.00
• The UltiMate™ 3000 with PAD system o
direct determination of simple carbohyd
an RSD of less than 2% (height) over a
method allows for detection of 100 pg s
• The use of the derivatizing agent 2AA a
sensitivity and selectivity for the measur
• The direct HILIC-charged aerosol detec
accurately and precisely determine sim
samples were easily processed by weig
mobile phase compatible solutions. Dire
separation provided adequate separatio
achieved using the Corona CAD. Using
can be performed using mobile phases
detectors. This helps to simplify analysi
enables confirmation of structures.
• The N- and O-linked glycans derived fro
glycoproteins can be profiled and chara
aerosol detection-MS. For glycoprotein
some of the pitfalls of PAD (high salt), a
impurities, ion-pairing agents). These is
offline-MS/MS analysis of unknowns. Th
instrument and thus amenable for use i
QC operations.
References
60
HILIC-mode
8.00
Conclusions
10.0
8.0
Glycopeptide #1
200ug/ml
25.0
4.00
0
AH08MAR0704
100
%
FIGURE 4. Carbohydrate measurement using HILIC-mode HPLC with charged
aerosol detection.
min
%
25
5 Carbohydrate Analysis using HPLC with PAD, FLD, Charged Aerosol Detection, and MS Detectors
1. Carlson D.M. Structures and immunoc
from pig submaxillary mucins. J. Biol. C
2. Huang, Y.; Mechref, Y.; Novotny, M.V.
glycans for subsequent analysis throug
electrophoresis. Anal Chem. 2001, 73,
3. Anumula KR., Advances in fluorescenc
liquid chromatographic analysis of glyc
350(1),1-23.
4. Technical Note 109: Analysis of 2-Amin
HPLC with Fluorescence Detection, LP
Scientific. http://www.dionex.com/en-u
Aminobenzamide-Glycans-09Sept201
Acknowledgements
Thermo Scientific would like to thank Dr. Andrew J.S. H
providing the data on LC-MS analysis of glycans. Dr Ha
fluorescence.
TSKtel is a registered trademark of Tosoh Corp. Shode
registered trademark of Showa Denko KK. All other tra
subsidiaries. This information is not intended to encour
the intellectual property rights of others.
FIGURE 6. Fetuin: reductive β-elimination: LC/Charged Aerosol Detection/
MS QTof (+ mode) using a Hypercarb PGC column with parallel charged aerosol
detection and MS.
Scientific.4 The
on accurate mass
rosol Detection
ymeric amino column
mns, provides adequate
proved selectivity and
hieved by decreasing
carbohydrates were
n (1 µg/mL using a
r ELSD. This method
amples (Figure 5).
hown) have also been
mine sugar abundance
of detecting analytes
al conditions and
%
900 uL to CAD
0
AH08MAR0704
100
TOF MS ES+
TIC
5.91e3
Glycopeptide #1
%
PLC with charged
0
•de-N glycosylate
•β-elimination
•desalt (evap. and AG-50)
•C18 cleanup
•LC/CAD/MS
Glycopeptide #2
0
100 uL to MS
8.00
10.00
12.00
14.00
16.00
18.00
20.00
22.00
24.00
26.00
28.00
30.00
32.00
34.00
36.00
Time
Conclusions
min
16.0
18.0
20.0
uice samples.
CAD_1
CAD_1
CAD_1
CAD_1
CAD_1
• The UltiMate™ 3000 with PAD system offers a routine and robust approach for the
direct determination of simple carbohydrates. The reproducibility of this method shows
an RSD of less than 2% (height) over a 14 hour period. The mass sensitivity of this
method allows for detection of 100 pg simple carbohydrates.
• The use of the derivatizing agent 2AA and fluorescence detection offers excellent
sensitivity and selectivity for the measurement of glycans from gycoproteins.
• The direct HILIC-charged aerosol detection approach offers a simple way to both
accurately and precisely determine simple carbohydrates. Fruit juices and food
samples were easily processed by weighing the sample and diluting the material in
mobile phase compatible solutions. Direct injection of samples using HILIC mode
separation provided adequate separation of sugars with low ng detection levels
achieved using the Corona CAD. Using this platform, useful product characterizations
can be performed using mobile phases which are compatible with nebulizer based
detectors. This helps to simplify analysis and the use of MS as an orthogonal detector
enables confirmation of structures.
• The N- and O-linked glycans derived from recombinant proteins and other
glycoproteins can be profiled and characterized using PGC RP-HPLC-Charged
aerosol detection-MS. For glycoprotein analysis, detection by the Corona CAD avoids
some of the pitfalls of PAD (high salt), and fluorescence (O-glycan peeling, fluorescent
impurities, ion-pairing agents). These issues can hamper LC-MS and downstream
offline-MS/MS analysis of unknowns. The Corona CAD is a simple and robust
instrument and thus amenable for use in routine development and even manufacturing
QC operations.
References
min
16.0
18.0
20.0
n
he two analytical
providing the profile of
uctural information. This
ombined technique is a
1. Carlson D.M. Structures and immunochemical properties of oligosaccharides isolated
from pig submaxillary mucins. J. Biol. Chem. 1968, 243, 616−626.
2. Huang, Y.; Mechref, Y.; Novotny, M.V. Microscale nonreductive release of O-linked
glycans for subsequent analysis through MALDI mass spectrometry and capillary
electrophoresis. Anal Chem. 2001, 73, 6063–6069.
3. Anumula KR., Advances in fluorescence derivatization methods for high-performance
liquid chromatographic analysis of glycoprotein carbohydrates. Anal Biochem. 2006,
350(1),1-23.
4. Technical Note 109: Analysis of 2-Aminobenzamide (AB) Labeled Glycans Using
HPLC with Fluorescence Detection, LPN2898, 2011. Dionex, Part of Thermo Fisher
Scientific. http://www.dionex.com/en-us/webdocs/111143-TN109-HPLC-2Aminobenzamide-Glycans-09Sept2011-LPN2898.pdf (January 30, 2012).
Acknowledgements
Thermo Scientific would like to thank Dr. Andrew J.S. Hanneman and Jason C. Rouse at Wyeth Pharmaceuticals for
providing the data on LC-MS analysis of glycans. Dr Hanneman also prepared and analyzed the glycans with
fluorescence.
TSKtel is a registered trademark of Tosoh Corp. Shodex is a trademark of Showa Denko KK. Asahipak is a
registered trademark of Showa Denko KK. All other trademarks are the property of Thermo Fisher Scientific and its
subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe
the intellectual property rights of others.
PO70026_E 2/12/S
6 Carbohydrate Analysis using HPLC with PAD, FLD, Charged Aerosol Detection, and MS Detectors
www.thermoscientific.com/dionex
Thermo Scientific Dionex products are
designed, developed, and manufactured
under an ISO 9001 Quality System.
TSKtel is a registered trademark of Tosoh Corp. Shodex is a trademark of Showa Denko KK. Asahipak is a registered trademark of Showa Denko KK.
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