2
CORTECS and CORTECS Premier 2.7
µ
m Columns
[ CARE AND USE MANUAL ]
II. GETTING STARTED
With each CORTECS Column, a Certificate of Analysis and
Performance Test Chromatogram are available in the column
box or on the column’s eCord Intelligent Chip. The Certificate
of Analysis is specific to each batch of packing material and
includes the batch number, analysis of unbonded particles,
analysis of bonded particles, and chromatographic results and
conditions. The Performance Test Chromatogram is specific
to each individual column and contains the batch number,
column serial number, USP tangent efficiency, USP tailing
factor, retention factor, and chromatographic conditions. These
data should be stored for future reference. For those not able
to access the information on the eCord Intelligent Chip, the
Certificate of Analysis and Performance Test Chromatogram
are available on request at www.waters.com/coa.
a. eCord Installation
(May not be available for all column configurations)
The eCord Intelligent Chip button is designed for use on
the ACQUITY Premier, Arc Premier, ACQUITY™ UPLC™ and
ACQUITY Arc™ Systems and it should be attached to the side
of the instruments’ column heater module. The eCord button
is magnetized and does not require specific orientation. For
more information on eCord Intelligent Chip functionality, see
Section IV in this care and use manual.
b. Column Installation
(with or without a VanGuard FIT Cartridge)
Note: Prior to handling CORTECS Columns and any chemical,
consult with your safety department and/or local regulations on
the use of proper protective equipment.
The CORTECS Columns are shipped in 100%
acetonitrile. The flow rates given in the procedure
below are for 2.1 mm I.D. columns.
1. Purge the pumping system of any buffer-containing
mobile phases and connect the inlet end of the column
to the injector outlet.
2. Flush column with 100% organic mobile phase (methanol
or acetonitrile) by setting the pump flow rate to 0.1 mL/min
and increase the flow rate to 0.5 mL/min over five minutes.
3. When the mobile phase is flowing freely from the column
outlet, stop the flow and attach the column outlet to the
detector. This prevents entry of air into the detection system.
4. Gradually increase the flow rate as described in Step 2.
5. Once a steady backpressure and baseline have been
achieved, proceed to the next section.
c. Column Equilibration
It is important to ensure mobile-phase compatibility before
changing to a different mobile-phase system. Equilibrate the
column with a minimum of 10-column volumes of the mobile
phase to be used (see Table 1 for a list of column volumes).
The column may be considered thermally equilibrated once
a constant backpressure is achieved.
Table 1. Empty Column Volumes in mL
(multiply by 10 for flush solvent volumes)
Internal Diameter
Column
Length (mm)
1.0 mm
2.1 mm
3.0 mm
4.6 mm
30
–
0.1
0.2
0.3
50
0.04
0.2
0.4
1.0
100
0.08
0.4
0.8
2.0
150
0.12
0.5
1.0
2.5
To avoid precipitating mobile-phase buffers on your column or
in your 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. (For example,
flush the column and system with 60% methanol in water prior
to introducing 60% methanol/40% buffer mobile-phase.)
For columns packed with HILIC material, 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 (see Table 1 for a list of column volumes). See
“Getting Started with HILIC Columns” for additional
information.
d. 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. The analyte mixture as found on the “Performance Test
Chromatogram” that 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
to achieve extremely low system volumes. This presents a
more challenging test of how well the column was packed,
and 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