Waters ACQUITY UPLC CSH Columns - Manual (Page 3)

[ Care and Use ManUal ]
ACQUITY UPLC CSH Columns 3
2. Determine the number of theoretical plates (N) and use this value for
periodic comparisons.
3. Repeat the test at predetermined intervals to track column perfor-
mance over time.
f. VanGuard Pre-Columns
VanGuard ™ Pre-columns are 2.1 mm i.d. x 5 mm length guard column
devices designed specifically for use in the ACQUITY UPLC systems.
VanGuard Pre-columns are packed with the same chemistries and frits
as our 2.1 mm i.d. ACQUITY UPLC CSH columns. VanGuard Pre-
columns are designed to be attached directly to the inlet side of an
ACQUITY UPLC CSH column.
Note: In order to ensure void-free and leak-free connections, the VanGuard Pre-column is
shipped with the collet and ferrule NOT permanently attached. Care must be taken when
removing the O-ring that holds these two pieces on the pre-column tubing.
ACQUITY UPLC ® Column VanGuard ™ Pre-Column
Place wrench here
Ferrule
Collet
Place wrench here
Flow
Installation Instructions
1. Remove VanGuard Pre-column from box and shipping tube and remove
plastic plug.
2. Orient pre-column so that male end is facing up and carefully remove
rubber O-ring that holds collet and ferrule in place during shipping
(collet and ferrule are not yet permanently attached).
3. Orient ACQUITY UPLC CSH column perpendicular to work surface so
that column inlet is on the bottom (column outlet on top).
4. From below, insert VanGuard Pre-column into ACQUITY UPLC
CSH column inlet and hand-tighten (collet and ferrule are not yet
permanently attached).
5. While pushing the VanGuard Pre-column into the column inlet,
turn assembled column and pre-column 180˚ so that pre-column
is now on top.
6. Tighten with two 5/16” wrenches placed onto ACQUITY UPLC CSH
column flats and VanGuard Pre-column hex nut (male end) as shown
above.
7. Tighten 1/4 turn to set collet and ferrule.
8. Check that ferrule is set by loosening connection and inspecting
the ferrule depth. A properly set ferrule depth will resemble other
connections in the ACQUITY UPLC system.
9. Reattach pre-column, apply mobile-phase flow and inspect for leaks.
II. CoLUMn Use
To ensure the continued high performance of ACQUITY UPLC CSH
columns, follow these guidelines:
a. Sample Preparation
1. Sample impurities often contribute to column contamination. One
option to avoid this is to use Oasis ® solid-phase extraction cartridges/
columns or Sep-Pak ® cartridges of the appropriate chemistry to clean
up the sample before analysis. For more information,
visit www.waters.com/sampleprep
2. It is preferable to prepare the sample in the operating mobile phase
or a mobile phase that is weaker than the mobile phase for the best
peak shape and sensitivity.
3. If the sample is not dissolved in the mobile phase, ensure that the
sample, solvent and mobile phases are miscible in order to avoid
sample and/or buffer precipitation.
4. Filter sample with 0.2 µm membranes to remove particulates. If the
sample is dissolved in a solvent that contains an organic modifier (e.g.,
acetonitrile, methanol, etc.) ensure that the membrane material does
not dissolve in the solvent. Contact the membrane manufacturer with
solvent compatibility questions. Alternatively, centrifugation for 20
minutes at 8000 rpm, followed by the transfer of the supernatant
liquid to an appropriate vial, could be considered.
b. pH Range
Please see Table 2 below for information on pH range and suggested
operating temperatures.