Waters XSelect CSH XP 2.5 µm Columns - Manual (Page 4)

XSelect CSH XP 2.5 µm Columns 4
[ CARE AND USE MANUAL ]
Table 3: Column Volumes (mL)
Column Length
(mm)
Internal Diameter
2.1 mm 3.0 mm 4.6 mm
30 0.10 0.21 0.50
50 0.17 0.35 0.83
75 0.26 0.53 1.25
100 0.35 0.71 1.66
To avoid precipitating mobile-phase buffers within the column or
system, flush the column with five column volumes of a water/organic
solvent mixture using the same, or lower, solvent content as in the
desired buffered mobile phase (i.e., flush the column and system with
60% methanol in water prior to introducing 60% methanol/40%
buffer mobile phase).
Note: If mobile-phase additives (i.e., ion-pairing reagents) are present in low
concentrations (<0.2% v/v), 100 to 200 column volumes may be required for
complete equilibration. In addition, mobile phases that contain formate (i.e.,
ammonium formate, formic acid) may require extended equilibration times.
g. eCord Installation
eCord ™ Technology represents a significant advancement in column
usage tracking management which can be realized if the column is
installed on an ACQUITY UPLC System. The eCord can be read
by connecting the yellow fob to the reader/writer located on the
right-hand side of the ACQUITY UPLC Column heater module.
Embedded information such as the column manufacturing QC data
and Certificates of Analysis may then be accessed via the ACQUITY
UPLC console.
h. Initial Column Efficiency Determination
1. Perform an efficiency test on the column before using it to track column
performance over time. This test may consist of:
a. An analyte test mixture that is commonly used in your laboratory
b. An analyte mixture as found on the “Performance Test
Chromatogram” which can be accessed via the eCord.
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 and expected. The Waters isocratic column testing systems have
been modified in order to achieve extremely low system dispersion. This presents
a more challenging test of how well the column was packed. This also guarantees
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 periodically to track column performance over
time. Slight variations may be obtained on different LC systems
due to the quality of the connections, operating environment,
system electronics, reagent quality, column condition and
operator technique.
i. VanGuard Pre-Columns
VanGuard ™ Pre-Columns are 2.1 mm ID x 5 mm length guard column
devices designed specifically to protect an analytical column while
minimizing the negative dispersion impact of utilizing such a device.
VanGuard Pre-Columns are packed with the same stationary phases
as the XP 2.5 µm Column offering. VanGuard Pre-Columns
are designed to be directly attached to the inlet of a eXtended
Performance [XP] 2.5 µm Column.
Note: VanGuard Pre-Columns are shipped with a collet and ferrule that are NOT
pre-swaged. This enables the end user to mate the VanGuard Pre-Column to a
specific XP 2.5 µm Column and ensures void-free and leak-free connections.
Care must be taken when removing the O-ring that holds these two pieces on
the pre-column tubing.
2.5 µm XP Column VanGuard Pre-Column
Place wrench here
Ferrule
Collet
Place wrench here
Flow
Figure 4: Installing a VanGuard Pre-Column.
VanGuard Pre-Column Installation Instructions
1. Remove the VanGuard Pre-Column from its box and shipping
tube and remove the plastic plug.
2. Orient the pre-column so that the male end is facing up and
carefully remove the black O-ring that holds the collet and
ferrule in place during shipment (collet and ferrule are not
permanently attached).