Waters ACQUITY UPC2 BEH, HSS, and CSH Columns - Manual (Page 2)

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[ CARE AND USE MANUAL ]
ACQUITY UPC 2 Torus Columns Care & Use
II. GET TING START ED
Each ACQUITY UPC 2 Column comes with a Certificate of Analysis
and a Performance Test Chromatogram. The Certificate of Analysis
is specific to each batch of packing material and includes the gel
batch number, physical characterization, analysis of unbonded
particle, analysis of bonded particles, and a SFC chromatographic
batch test. The Performance Test Chromatogram is specific to
each individual column and contains the following information:
gel batch number, column serial number, USP plate count, USP
tailing factor, capacity factor, and chromatographic conditions
under normal-phase LC conditions. These data should be stored for
future reference.
a. Safety Considerations
An SFC column, while in use, is under pressure with CO
2
and
possible modifiers as a supercritical fluid. A major safety concern
is frostbite caused by adiabatic cooling when CO
2
decompresses
from a fluid to a gas at atmospheric pressure. Pay attention to any
frosting on the column or system connections. This indicates a
leak, usually with temperatures far below 0 °C.
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 volumes. For information on system tubing and
connectors, please refer to the ACQUITY UPC
2 System Operator’s
Guide (Part Number 720004226EN).
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 (typically methanol) as organic modifier.
2. Connect both the inlet and outlet of the column to the
SFC 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.)
e. Initial Column Efficiency Determination
1. Perform an efficiency test on the column before using it.
Waters recommends using a suitable solute mixture to analyze
the column upon receipt. We recommend that weaker solvents
are selected as injection solvents. Peak distortion can occur
due to strong solvent effects, resulting in lower efficiency
values. (Methanol can be considered a strong solvent under
SFC conditions.)
Table 1. Physical Characteristics.
Chemistry
Particle
Shape
Particle
Size (µm)
Pore
Size (Å)
Surface Area
(m2/g)
Pore Volume
(cc/g)
Carbon
Load (%)
Endcapped
Particle
Technology
BEH 2-Ethylpyridine (2-EP) Spherical 1.7, 3.5 130 185 0.7 9 No BEH
BEH (Unbonded) Spherical 1.7, 3.5 130 185 0.7 N/A N/A BEH
CSH Fluoro-Phenyl Spherical 1.7, 3.5 130 185 0.7 10 No CSH
HSS C
18
SB Spherical 1.8, 3.5 100 230 0.7 8 No HSS