Waters CORTECS 2.7 um Columns - Manual (Page 4)

4 CORTECS 2.7 µm Columns
Table 3. Empty Column Volumes (mL)
Internal Column Diameter
Column Length (mm) 1.0 mm 2.1 mm 3.0 mm 4.6 mm
30 0.024 0.10 0.21 0.50
50 0.039 0.17 0.35 0.83
75 0.059 0.26 0.53 1.25
100 0.079 0.35 0.71 1.66
150 0.12 0.53 1.07 2.49
For CORTECS HILIC Columns, flush with 50 column volumes of
50:50 acetonitrile/water with 10 mM final buffer concentration.
Prior to the first injection, equilibrate with 20 column volumes
of initial mobile-phase conditions (refer to Table 1 for a list of
column volumes). If gradient conditions are used, equilibrate
with 8–10 column volumes between injections. Failure to
appropriately equilibrate the column could results in drifting
retention times. See “Getting Started with CORTECS HILIC
Columns” under “Additional Information”.
g. eCord Installation
eCord Technology represents a significant advancement in column
usage tracking management when connected to an ACQUITY UPLC
System. The eCord Intelligent Chip 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 be accessed via the ACQUITY UPLC console. The
eCord is available on all 2.1 mm, and 3.0 mm column dimensions.
h. Functional Tests for Benchmarking a New Column
Waters recommends performing a benchmarking test upon receipt
of your column and throughout lifetime usage. By using a standard
mix such as the appropriate Quality Control Reference Material
(QCRM), you can:
■ ■
Verify the performance of your column upon receipt
■ ■
Monitor the health of your column and system over time
■ ■
Troubleshoot separation difficulties that arise
The Neutrals QCRM (p/n 186006360) or HILIC QCRM (p/n
186007226) are appropriate mixtures with which to benchmark
your CORTECS Column and System. Other QCRMs are available
depending on the application and detection mode.
1. Run an initial benchmark test to monitor key performance
criteria such as retention time, peak area, peak tailing,
resolution, response, system pressure, etc.
2. Repeat test daily to track column and system performance
over time.
For more information on benchmarking performance, visit
www.waters.com/QCRM
Note. If an analyte mixture found in the Performance Test
Chromatogram is used to benchmark, 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.
3. Determine the number of theoretical plates (N) and use this
value for periodic comparisons.
4. 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 designed to be attached directly to the
inlet of a CORTECS Analytical Column. They are 2.1 mm i.d. x 5 mm
length guard columns packed with CORTECS solid-core silica
particles. The chemistries offered for VanGuard Pre-Columns are
identical to the available CORTECS chemistries.
Note. VanGuard Pre-Columns are shipped with a collet and ferrule that
are NOT pre-swaged. This enables the end user to seat the VanGuard
Pre-Column to the specific CORTECS Column to ensure 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.
CORTECS Column VanGuard Pre-Column
Place wrench here
Ferrule
Collet
Place wrench here
Flow
Figure 4. Installing a VanGuard Pre-Column.