Waters UPLC Intact Mass Analysis Application Kit User Manual
[ CARE AND USE MANUAL ]
UPLC Intact MASS Analysis Application Kit
(Part Number: 176001519)
CONTENTS
I. UPLC INTACT MASS ANALYSIS APPLICATION
KIT COMPONENTS
II. INSTALLING THE PRE-COLUMN TUBING INTO
MASSPREP MICRO DESALTING COLUMN
III. CONNECTING THE COLUMN TO LCT
PREMIER MS
IV. CONNECTING THE COLUMN TO Q-Tof
PREMIER MS, AND SYNAPT HDMS
V. CONNECTING THE COLUMN TO WATERS SQ
DETECTOR (SQD)
IV. SUGGESTED METHOD FOR MASS ANALYSIS
USING UPLC INTACT MASS ANALYSIS
APPLICATION KIT
a. Intact Antibody Analysis
b. Reduced Antibody Analysis
I. UPLC INTACT MASS ANALYSIS APPLICATION
KIT COMPONENTS
Description Qty Part Number
MassPREP Micro Desalting Column 1 186004032
Desalting Cartridge Manual Assembly,
Cap/Tube/Column
UPLC Intact Mass Analysis Solution 1 715001664
Intact Mass Tubing Kit. Kit contains: 1 205000548
Peek Compression Screw, Long, 10–32 4 410002276
Peek Tubing, 0.062 OD x .004 ID x 8.50 L 1 430001991
Peek Tubing, 0.062 OD x .004 ID x 15.04 L 1 430001992
Peek Tubing, 0.062 OD x .004 ID x 22.20 L 1 430001993
P/N: 430001988
P/N: 410002276
P/N: 186004032
1 430001988
VII. TIPS FOR SUCCESS
P/N: 430001991
P/N: 430001992
P/N: 430001993
Figure 1. UPLC® Intact Mass Analysis Application Kit Components.
[ CARE AND USE MANUAL ]
II. INSTALLING THE PRE-COLUMN TUBING INTO
MASSPREP MICRO DESALTING COLUMN
Required materials:
1/4-inch open-end wrench
5/16-inch open-end wrench
Flow
Solvent Inlet Solvent Outlet
Figure 3. Column assembly after attaching the column to the pre-column tubing.
III. CONNECTING THE COLUMN TO LCT
PREMIER MS
Figure 2. Installation of the pre-column tubing into the MassPREP Micro
Desalting Column.
Installation Procedure
1. Place the column on a flat surface (O-ring end facing up).
2. Remove the O-ring from the column.
3. Secure the column (P/N 186004032) with a 5/16-inch wrench
(bottom) as shown in Figure 2.
4. Tighten one end of the pre-column tubing (P/N 430001988)
into the column with a 1/4-inch wrench (a full turn beyond
hand-tight).
5. Connect the marked end (marked as solvent inlet) of the
column tubing assembly to the solvent outlet of the ACQUITY
UPLC® Column Heater (Figure 3).
Note: Proceed from this point to either section III, IV, or V
depending upon the type of mass spectrometer you have.
Installation Procedure
1. Connect one end of the 15.04-inch long PEEK tube (P/N
430001992) to the outlet of the column using a fitting (P/N
410002276).
2. Connect the other end of the tubing to the salt divert valve
(Port 2) using another fitting, as shown in the Figure 4. The
salt divert valve is located at the top-left corner of the LCT
Premier™ front panel.
3. Place the column in the column heater as shown in the Figure 6.
4. Connect one end of the 8.5-inch long PEEK tube (P/N
430001991) to Port 1 of the divert valve using a fitting
5. Connect the other end to the inlet of LCT Premier sample probe
using another fitting (Figure 4).
6. Use a larger diameter PEEK tube (0.062 o.d. x 0.005 or
0.01-inch i.d.) for the waste line (not included in the kit).
UPLC Intact MASS Analysis Application Kit
2
[ CARE AND USE MANUAL ]
Column Heater
UPLC
Gradient
Waste
Position 1: Sample loading and salt divert to waste
UPLC
Gradient
Waste
Position 2: Sample elution to MS
Figure 4. Fluidic configuration for LC-MS (UPLC-LCT Premier) analysis. A postcolumn salt diversion valve (top-left corner of the LCT Premier) can be utilized
to divert buffers and nonvolatile salts to waste in the beginning of the LC/MS
analysis.
15.04”
8.5”
15.04”
8.5”
IV. CONNECTING THE COLUMN TO Q-TOF Premier
MS, AND SYNAPT HDMS
Neither Q-Tof Premier™ MS nor SYNAPT® HDMS® has an integrated
divert valve. However, an optional external 2 position valve (P/N
MS
MS
417000118) can be used with Q-Tof Premier and Synapt HDMS
mass spectrometers to divert buffers and nonvolatile salts present
in the sample to waste at the beginning of the LC/MS analysis.
Follow Steps 1–5 in Section II to attach the column to the pre
column tubing. Then follow Steps 12-17 below to connect the
column to the mass spectrometer.
1. Connect one end of the 15.04-inch long PEEK tube
(P/N 430001992) to the outlet of the column using a fitting
(P/N 410002276).
2. Connect the other end of the tubing to the external salt divert
valve (Port 4) using another fitting, as shown in the Figure 5.
3. Connect one end of the 8.5-inch long PEEK tube (P/N
430001991) to the external divert valve (Port 5).
4. Connect the other end to the inlet of MS sample probe (Figure 5).
The divert valve can be programmed from the MS Method Editor in
MassLynx®. To set a salt divert time, open MS Method Editor. Then
select Option> Solvent Delay on the MS Method Editor to open
the Solvent Delay dialog box. To enable the divert/injection valve
to be used as divert valve, select Enable Divert Valve. This diverts
the flow of solvent during a solvent delay period (0.5 min or user
defined value) either to, or away from, the source for the time
period shown in the solvent delay timetable (Table 1).
5. Place the column in the column heater as shown in the Figure 6.
6. Use a larger diameter PEEK tube (0.062 o.d. x 0.005 or
0.01-inch i.d.) for the waste line (not included in the kit).
Table 1. Salt divert timetable.
UPLC Intact MASS Analysis Application Kit
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