Waters Sugar-Pak I Column - Manual (Page 2)

[ Care and Use ManUal ]
Sugar-Pak I Columns 2
II. PRePARAtIon FoR oPeRAtIon
a. Column Installation
Remove the end plugs from the steel column with a 5/16-inch open-
end wrench and save them for storage when the column is removed
from the system. The column outlet is indicated by an arrow on the
label (showing the direction solvent should flow). Tighten the fittings
to turn past finger tight. DO NOT OVERTIGHTEN – this will damage the
fitting seat. A properly prepared and assembled compression fitting in
good condition is all that is required.
If tube cutting is required to connect a new column or to improve the
end connections on your existing fittings, follow these steps:
1. Use a file with a cutting edge (such as the file included in the
Startup Tool Kit, P/N WAT096146) to scribe the circumference of
the tube at the desired break.
2. Grasp the tube on both sides of the scribe mark with cloth-
covered or smooth-faced pliers (to prevent marring the tube
surface) and gently work the tube back-and-forth until it
separates.
Figure 1. Ferrule and compression assembly.
3. Inspect the tube at the break for burrs. File the outer edges at an
angle to the tube opening. Do not file flat across the open tube
as this might cause plugging or uneven flow delivery. Assemble
as shown.
4. Slide the compression fitting over the tube, followed by the
ferrule (large end of the taper first).
5. Seat the ferrule by tightly mating the assembly to the fitting seat
in which it will be used. An improperly positioned ferrule can
form unwanted dead volume which could result in unintentional
sample mixing.
Note: Attach a union in place of the column and flush the lines
free of microparticulates before attaching the column.
b. Mobile-Phase Requirements
This column is packed with a calcium-loaded resin. Hydrogen or
other cations can replace the calcium and cause inversion on the
column of sugars such as sucrose, which are prone to inversion.
It is recommended that a small amount of calcium be used in the
mobile phase to maintain the equilibrium and prevent inversion.
The recommended mobile phase is deionized, bacteria-free water
containing approximately 0.0001 M calcium EDTA* (50 mg/L).
* Calcium EDTA has several common names:
- Calcium di-sodium (ethylene dinitrilo) tetra-acetic acid
- Calcium di-sodium ethylene diamine tetraacetate
- Calcium di-sodium edentate
 Water should be deionized to greater than 2 megohms resistivity.
It is essential that the water used is free of polyvalent cations,
particularly transition and heavy metals (e.g., iron).
 Remove bacteria and other particulates from the water just
before use by vacuum filtration. The Solvent Clarification Kit is
recommended for solvent filtration (110 V, P/N WAT085113;
240 V, P/N WAT085122) using PES filter membranes (P/N
WAT200538).
 Although the mobile phase will be partially degassed by vacuum
filtration, it is highly desirable that it be thoroughly degassed by
the following procedure:
1. Place the mobile phase in an Erlenmeyer flask on a
stirrer/hot plate.
2. Cover the mouth of the flask with aluminum foil to minimize
evaporation.
3. Bring the mobile phase to its boiling point for a few
minutes just before use, but maintain the temperature
between 70-90 °C during use. This practice ensures that
gases (especially C0
2
) do not redissolve in the mobile
phase. It also prevents the growth of microorganisms.
4. Keep the mobile-phase reservoir clean and covered and supply
freshly prepared mobile phase every 24 hours.
 If the system is to be used continually for more than one
day, place the mobile phase in an Erlenmeyer flask on a
stirrer/hot plate. Cover the mouth of the flask with aluminum
COMPRESSION SCREW OR NUT
TUBE
FERRULE
END MUST BE STRAIGHT
AND SMOOTH TO ACHIEVE
MAXIMUM COLUMN EFFICIENCY
CRTITICAL DISTANCE TO BE DETERMINED BY
EACH APPLICATION (UNION, COLUMN FITTING, ETC.)