Waters Protein-Pak Hi Res IEX Columns - Manual (Page 2)

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[ CARE AND USE MANUAL ]
Protein-Pak Hi Res IEX Columns and Standards
II. CONFIGURING AN ACQUIT Y SYST EM FOR
USE IN IEX P ROT EIN SEPARATIONS
Although Waters Protein-Pak Hi Res IEX columns do not require
the high pressure capability of an ACQUITY UPLC system, the full
performance benefits of this column will only be realized with the
low system volume and low detector dispersion of the optimized
ACQUITY UPLC system. However, ion-exchange chromatography
may require modifications to an existing ACQUITY UPLC system.
Please refer to “Size Exclusion and Ion-Exchange Chromatography
of Proteins using the ACQUITY UPLC System”, (715002147A) for
specific recommendations that can be obtained at www.waters.
com/chemcu.
III. GET TING START ED
An Analysis Report and an Inspection Data Sheet are generated
and available for each shipped Protein-Pak Hi Res IEX column.
The Analysis Report is specific to each batch of packing material
and includes the batch number and chromatographic separation
obtained using defined protein standards. This document is
available upon request http://www.waters.com/waters/form.
htm?id=10048430&ev=10048691&locale=en_US. The
Inspection Data Sheet is included with each shipment and is
specific to each individual column and contains the following
information: batch number, column serial number, plate count,
tailing factor, retention factor, and chromatographic separation
conditions. These data should be stored for future reference.
a. eCord Installation
The eCord™ button should be attached to the side of the Waters
ACQUITY UPLC column heater module. The eCord button is
magnetized and does not require specific orientation.
b. Column Connectors
There are two options for connecting the Protein-Pak Hi Res
column to an ACQUITY system. The first option is to use the
reusable fitting on the 150 mm and 100 mm column stabilizers
(sect b1). This fitting/tubing is supplied with ACQUITY UPLC
column heaters. A second alternative is to remove this fitting and
replace it with a non removable fitting (i.e. steel). If the second
option is employed, specific directions must be followed (sec b2).
b1. The ACQUITY UPLC system utilizes tubing and gold plated
compression screws that have been designed to meet
stringent tolerance levels and to minimize extra column
volumes. The columns should be attached to the ACQUITY
UPLC injector with a column stabilizer.
b2. Waters Protein-Pak Hi Res columns require end fittings that
have a 2 mm depth between the ferrule and the end of the
extending stainless steel tubing (Figure A) that must perfectly
seat to the end of the Protein-Pak Hi Res column. Connecting
a different style connector to a Waters Protein-Pak Hi Res
IEX column may leave a gap between the end of the stainless
steel tubing and the column that will result in undesired peak
broadening (Figure B).
To correct this problem, disassemble the finger-tight fitting of
the stabilizer and remake the connection (i.e., 2 mm depth) to the
Waters Protein-Pak Hi Res IEX column.
Figure A
Figure B
In a proper tubing/column connection, the tubing touches the
bottom of the column end fitting with no void between them.
(Figure C).
Figure C
2 mm