Waters Gen-Pak FAX Columns - Manual (Page 2)

[ Care and Use ManUal ]
Gen-Pak FAX Columns 2
II. InstALLAtIon
a. Column
Before you attach a new column, attach a union in place of the column and
flush the HPLC system free of previously used solvent.
Note: To achieve optimum performance from each column, use tubing of
0.010 inch i.d. or smaller to connect the injector to the column and the
column to the detector.
Procedure to install the column:
1. Remove the end plugs from the column. (Be sure to save the end
plugs for use when the column is removed from the system for
storage.) The column outlet is indicated by an arrow on the label
showing the direction solvent should flow.
2. Finger-tighten the tubing compression screws and then
wrench-tighten them 1/4 to 1/2 turn.
Note: Do not over-tighten; this damages the connection.
3. Make sure that the compression fitting is in good condition and
properly prepared as shown in Figure 1. Because fittings may
vary, it is important to verify that the tubing in your system
bottoms in the column end nuts.
Figure 1: Ferrule and Compression Screw Assembly
b. Tubing
Follow the next three steps to cut tubing to connect a new steel
column or to improve the end connections on existing fittings:
1. Using a three-cornered file with a cutting edge, scribe the
circumference of the tubing at the desired break.
2. Grasp the tubing on both sides of the scribe mark with
cloth-covered pliers (to prevent marring the tube surface) and
gently work the tube back and forth until it separates.
3. Slide the compression screw, followed by the ferrule (large end
of the taper first) over the tube. Be certain to bottom the tube in
the fitting seat to assure a leak-free connection.
c. Testing
To validate system and column performance, test each new column
with a standard sample. This provides a basis for detecting system
component changes and troubleshooting.
Suggested samples include:
• Hae Ill digest of ØX 174 RF DNA
• BstN I digest of pBR322 DNA
III. GeneRAL UsAGe ConsIDeRAtIons
This section presents guidelines that can extend the life of your
column and help you achieve the best possible chromatographic
results.
a. Samples
When preparing and using samples:
• Centrifuge or filter the sample before injection.
• Do not inject samples containing microparticulates.
b. Solvents
When preparing and using solvents:
• Use pure buffer salts.
• Use of high quality reagents, water, and solvents is critical in
preparing chromatography eluents. Fouling of the Gen-Pak FAX
resin, leading to a loss in retention and / or separation efficiency,
occurs faster on this column chemistry due to the small surface
area of the non-porous resin particles. As such, all prepared HPLC
eluents should be filtered through a solvent compatible 0.45 µm or
0.22 µm depth filter.
• Solvents should be degassed (vacuum filtration, sonication or
helium sparged) prior to use.
• Use of organic solvents other than methanol or acetonitrile at
greater than 10% in water is not recommended.
Tube
Critical distance to be determined by each
application (union, column fitting, etc.)
Compression screw or nut
Ferrule
End must be straight and smooth
to achieve maximum column
efficiency