Waters ACQUITY UPLC HSS Columns - Manual (Page 3)

[ Care and Use ManUal ]
XSelect HSS HPlc columnS 3
e. Initial Column efficiency Determination
1. Perform an efficiency test on the column before using it. this
test may consist of:
a) An analyte test mixture that is commonly used in your
laboratory, and/or
b) An analyte mixture as found on the “Performance test
chromatogram” which accompanied your column.
Note: If b) is performed, the isocratic efficiencies measured in
your laboratory may be less than those given on the Waters
“Performance Test Chromatogram”. This is normal. The Waters
isocratic column testing systems have been modified in order
to achieve extremely low system volumes. This presents a more
challenging test of how well the column was packed. This guar-
antees the highest quality packed column. These special testing
systems have been modified to such an extent that they are not
commercially viable and have limited method flexibility other
than isocratic column testing.
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
performance over time. Slight variations may be obtained on
two different uPlc
® systems due to the quality of the con-
nections, operating environment, system electronics, reagent
quality, column condition and operator technique.
f. VanGuard Pre-Columns
VanGuard Pre-columns are 2.1 mm ID x 5 mm length guard col-
umn devices designed specifically for use in the AcQuItY uPlc
system. VanGuard Pre-columns are packed with the same uPlc
chemistries and frits as our 2.1 mm ID uPlc columns. VanGuard
Pre-columns are designed to be attached directly to the inlet side
of an AcQuItY uPlc 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 the VanGuard Pre-column from its box and shipping
tube and remove plastic plug.
2. orient the 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 the AcQuItY uPlc column perpendicular to the work
surface so that column inlet is on the bottom (column outlet
on top).
4. From below, insert the VanGuard Pre-column into the
AcQuItY uPlc 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 the AcQuItY
uPlc column flats and the VanGuard Pre-column hex nut
(male end) as shown above.
7. tighten 1/4 turn to set collet and ferrule.
8. check that the ferrule is set by loosening the 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.