Waters ACQUITY UPC2 Trefoil Columns - Manual (Page 3)

3
[ CARE AND USE MANUAL ]
ACQUITY UPC 2 Trefoil Columns Care & Use
Any small leak could produce a situation where the LEL (lower
exposure limit) is reached. Laboratories should be equipped with
CO
2
and/or O
2
sensors when carbon dioxide (CO
2
) is in use.
b. Column Connectors and Installation
ACQUITY UPC 2 Systems utilize tubing and connectors which
have been designed to meet stringent tolerance levels and to
minimize extra-column volume. For information on system tubing
and connectors, please refer to the ACQUITY UPC
2 System
Operator’s Guide (Part Number 720004226EN can be found
on
www.waters.com/chemcu ).
Note: Scale the flow rate up or down accordingly based upon
the column i.d., length, particle size, and backpressure of the
ACQUITY UPC
2 Column being installed.
1. Make sure your co-solvent pump is primed and has an adequate
solvent/modifier supply before performing injections. It is
recommended to use CO
2
with purity level of 99.97% (food
grade) and use high-quality chromatography grade solvents
(see section IIIb).
2. Connect both the inlet and outlet of the column to the
UPC 2 System.
3. If the column is still filled with a solvent, use a low flow rate
and back pressure setting (100 bar) to start pumping CO
2
and
modifier through the column.
4. If you see frosting on the column at the inlet or outlet, tighten
the finger-tight fitting or compression screw on that side. If you
continue to see frosting, turn off the CO
2
and vent the system.
Allow the column to depressurize fully before disconnecting
the inlet or outlet to troubleshoot the leaking issue.
c. 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.
d. Column Equilibration
ACQUITY UPC 2 Columns are shipped dry. Equilibrate the column
with a minimum of 10-column volumes of the mobile phase prior
to use. (Refer to Table 2 for a listing of empty column volumes.)
Thermal and chemical equilibration are equally important
chromatography. Ensure that the mobile phase is preheated before
entering the column to prevent a thermal miss-match. This can be
done by enabling active preheating on a UPC
2 ® instrument.
e. Initial Column Efficiency Determination
1. Before use, perform an efficiency test on the column,
using suitable analytes dissolved in a weaker injection solvent
to avoid peak distortion from strong injection solvent effects
(see section IIIb). It is recommended to use Waters Quality
Control Reference Material (QCRM) (p/n 186007950)
which contains a mixture of standards specifically chosen
for the ACQUITY UPC
2 Trefoil and Torus Columns. For more
information, please review the ACQUITY UPC 2 Columns
Brochure (literature code 720004275EN).
ACQUITY UPC 2 Quality Control Reference Materials (QCRM)
Intended Use Contents
Provides
convergence
chromatographic
performance
information for
both chiral and
achiral modes 1. 0.50 mg/mL (+/-) trans-Stilbene oxide
2. 0.50 mg/mL Thymine
3. 0.50 mg/mL Sulfamethoxazole
4. 0.50 mg/mL Sulfamethizole
In a 1 mL solution of 75:25 ACN:MeOH
2. Determine the number of theoretical plates (N) and use this
value as a benchmark for periodic comparisons.
3. Repeat the test at predetermined intervals to track
column performance over time. (Slight variations may
occur if performance tests are performed on two different
ACQUITY UPC
2 Systems due to the quality of the connections,
operating environment, system electronics, reagent quality
and column condition.)
Empty Column
Volumes (mL)
Column Internal Diameter (mm)
Column Length 2.1 mm 3.0 mm
50 mm 0.2 0.4
150 mm 0.5 1.0
Table 1. Empty Column Volumes in mL (multiply by 10 for flush
solvent volumes).