Waters CORTECS 1.6 um Columns - Manual (Page 2)

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CORTECS Columns
I. GettInG stARteD
Each CORTECS Column comes with a Certificate of Analysis and
a Performance Test Chromatogram embedded within the eCord
intelligent chip. The Certificate of Analysis is specific to each batch
of packing material contained in the CORTECS Column and includes
the gel 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 such information as: gel batch number,
column serial number, USP plate count, USP tailing factor,
capacity factor, and chromatographic conditions. These data
should be stored for future reference.
a. Column Connectors
The ACQUITY UPLC System utilizes tubing and gold plated
compression screws which have been designed to meet stringent
tolerance levels and to minimize extra column volumes.
Optimized column inlet tubing (Part Number 430001084) is supplied
with the ACQUITY UPLC System. The inject valve end of the tubing is
clearly marked with a blue shrink tube marker. Insert the opposite end
of the tubing into the CORTECS Column and tighten the compression
fitting using two 5/16-inch wrenches.
For information on the correct column outlet tubing, please refer
to the relevant detector section in the ACQUITY UPLC System
Operator’s Guide (Part Number 71500082502).
b. Column Installation
Note: The flow rates given in the procedure below are for a typical
1.6 µm, 2.1 mm ID by 50 mm length column. Scale the flow rate
up or down accordingly based upon the column pressure and
system limits.
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 5 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
and gives more rapid baseline equilibration.
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.
Note: If mobile-phase additives are present in low concentrations
(e.g., ion-pairing reagents), 100 to 200 column volumes may be
required for complete equilibration. In addition, mobile phases that
contain formate (e.g., ammonium formate, formic acid, etc.) may
also require longer initial column equilibration times.
c. Column equilibration
CORTECS Columns are shipped in 100% acetonitrile. 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 (refer to Table 1 for
a list of column volumes). The column may be considered thermally
equilibrated once a constant backpressure is achieved.
table 1. Column Volumes (mL)
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 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). See “Getting Started with CORTECS HILIC Columns” for
additional information.
d. eCord Installation
The eCord button should be attached to the side of the column heater
module. The eCord button is magnetized and does not require specific
orientation.
Column length (mm)
Internal diameter
2.1 mm 3.0 mm
30 0.1 0.2
50 0.2 0.4
75 0.3 0.5
100 0.4 0.8
150 0.5 1.0