Waters MassPREP Online Desalting Cartridge User Manual
[ CARE AND USE MANUAL ]
MassPREP On-Line Desalting Cartridge
CONTENTS
I. INTRODUCTION
II. INSTALLING THE MASSP REP ON-LINE
DESALTING CART RIDGE INTO T HE SENTRY
2.1 X 10 MM GUARD COLUMN HOLDER
III. RECOMMENDED LC/MS SYSTEM CONFIGURATION
TO MINIMIZE MS SOURCE CONTAMINATION
IV. SUGGEST ED METHOD FOR MASS ANALYSIS
USING MASSPREP ON-LINE DESALTING
CARTRIDGE
a. Instrumentation
b. Separation Method
V. GUIDELINES FOR USE AND
TROUBLESHOOTING
VI. MISCELLANEOUS
I. INTRODUCTION
Mass spectrometry is a powerful tool for intact protein analysis.
However, the quality of analyses can be compromised by salts in
the sample that interfere with ionization and/or form adducts with
the investigated proteins. It is therefore important to remove or
significantly minimize the introduction of extraneous salt ions
into the mass analyzer. The use of on-line, desalting devices can
efficiently perform this function with minimal sample handling.
Waters MassPREP™ On-line Desalting Cartridges (2.1 x 10 mm,
P/N 186002785) contain a polymer-based, reversed-phase
packing material that can be used to effectively desalt protein
samples prior to mass spectrometry. (Note: The purchase and use
of Waters Sentry™ 2.1 x 10 mm Guard Column Holder Kit (P/N
WAT097958) is required for this application as shown in Figure 1.)
The phenyl phase material contained in these on-line devices
successfully “traps” proteins allowing the salts to be washed
to waste prior to elution of the desalted protein into the mass
spectrometer. With an optimized LC/MS method, cycle times as low
as 3 minutes are achievable. These on-line devices, when used with
an appropriate HPLC System injector and injector wash solution,
can be used for the sequential analyses of different samples (See
Figures 2 and 3).
This Care and Use manual provides suggested methods for the
successfully use of this product. For the latest list of MassPREP,
BioSuite™ or NanoEase™ column offerings, go to www.waters.com/
lifesciences.
II. INSTALLING THE MASSPREP ON-LINE
DESALTING CART RIDGE INTO T HE SENTRY
2.1 X 10 MM GUARD COLUMN HOLDER
1. Unscrew the two halves of the Sentry 2.1 x 10 mm Guard
Column Holder.
2. Remove the old MassPREP On-line Desalting Cartridge, if
present.
[ CARE AND USE MANUAL ]
3. Install the new MassPREP On-line Desalting Cartridge, noting
the flow arrow direction on the device. This should match the
flow arrow direction on the analytical column.
4. Reassemble the Sentry 2.1 x 10 mm Guard Column Holder.
Hand-tighten only.
5. Connect the 2.1 x 10 mm Guard Column Holder containing
the MassPREP On-line Desalting Cartridge onto HPLC or
LC/MS System using appropriate connectors (e.g., PEEK™
tubing and connectors). Prior to use, the MassPREP On-line
Desalting Cartridge reversed-phase packing material must
be “wet” with organic solvent prior to equilibration with 95%
Eluent A/5% Eluent B. This can be accomplished by running
a “blank injection” (i.e., buffer injection followed by running
the separations gradient) or by washing the column with a
minimum of five column volumes of 50% Eluent B followed by
a ten column volume equilibration at 95% Eluent A/5% Eluent
B prior to sample injection. While equilibrating, check the
holder and all connections for leaks.
III. RECOMMENDED LC/MS SYSTEM
CONFIGURATION TO MINIMIZE MS SOURCE
CONTAMINATION
The presence of non-volatile salts (e.g., NaCl) can suppress protein
ionization and foul the ionization source of the mass spectometer.
It is therefore important to remove or significantly minimize the
introduction of these compounds into the mass analyzer. Figure
2 details an LC/MS configuration that can effectively be used to
load sample onto the column and elute salt to waste prior to valve
switching and protein elution into the mass analyzer.
Position 1: Sample Loading and Salt Elution to Waste
Spectrometer
MassPREP
On-Line
Desalting
Cartridge
Waste
1
2
3
6
5
4
Gradient
HPLC
Mass
Figure 1: Sentry Guard Holder (P/N WAT097958) and MassPREP On-line
Desalting Cartridge (P/N 186002785).
HPLC
Pump
Recirculation
Position 2: Sample Elution to Mass Spec
Spectrometer
MassPREP
On-Line
Desalting
Cartridge
HPLC
Pump
Waste
1
2
6
5
4
3
Recirculation
Gradient
HPLC
Mass
Figure 2: LC/MS Valve Switching Configuration for use with MassPREP
On-line Desalting Cartridge.
MassPREP On-Line Desalting Cartridge
2
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