Waters Pico-Tag Amino Acid Analysis Column - Manual (Page 2)

[ Care and Use ManUal ]
Pico•Tag Amino Acid Analysis Column 2
II. PrePArAtIon For oPerAtIon
a. Column Installation
Remove the end plugs from your steel column with a 5/16” wrench and save
them for storage after the column has been removed from the system. The
column outlet is indicated by an arrow on the label (show-ing the direction
solvent should flow). Tighten the fittings 1/4-to-1/2 turn beyond finger tight.
DO NOT OVERTIGHTEN - THIS WILL DAMAGE THE FITTING SEAT. A properly
prepared and assembled com-pression fitting in good condition is all that
is required.
Follow the next four steps of this procedure if tubing cutting is required to
connect a new column or to improve the end connections on your existing
fittings.
1. Using a file with a cutting edge (such as the file included in the Startup
Tool Kit, P/N 96146, supplied with each Waters Liquid Chromatography
System) 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. File the ends smooth and assemble as shown.
4. Slide the compression fitting, followed by the ferrule (large end of the
taper first) over the tube. Be certain to bottom the tube in the fitting
seat for which its use is intended to assure a leak-free connection.
Note: Attach a union in place of the column and flush the lines free of
microparticulates before attaching the column
.
Figure 5: Ferrule and Compression Assembly
b. Solvent Requirements
Your Pico•Tag column is shipped containing an aqueous/organic mixture
compatible with the Pico•Tag eluents. It is recommended that only Waters
Pico•Tag eluents be used with this column.
Waters Pico•Tag Eluent A (P/N 88108)
Waters Pico•Tag Eluent B (P/N 88112)
Waters Pico•Tag Diluent (P/N 88119)
c. Equilibration
A necessary step to successful use of your column is the initial solvation
(or wetting) of the packing. Install the column in a column heater and bring
temperature to 38 °C. Purge the column at 1 ml/min with Eluent B, then
equilibrate with Eluent A for fifteen minutes. Equilibration between the mobile
phase and packing is established when a stable baseline can be produced. If
your result is un-satisfactory repeat the equilibration process.
III. cAre And use
Liquid chromatography columns have a finite life influenced by their care and
use, number of injections, sample and solvent cleanliness, frequency of sol-
vent changeover, and handling and storage procedures among other factors. If
a change is observed in the:
Retention of a particular compound resolution between two compounds peak
shape.
Take immediate steps to determine the reason for the changes, and until the
determination is made, the results of any separations using the column must
not be relied upon. Follow generally accepted pro-cedures for quality control
and methods development when using these columns.
Important Note: Before running the first analysis on your new column perform
the Test Sample Separation given in the Test Conditions Section.