Waters BioSuite Columns - Manual (Page 6)

[ Care and Use ManUal ]
BioSuite Columns 6
concentrations. Prior to chromatography, test to ensure the sample
does not precipitate at the organic concentration to be used for the
chromatography.)
6. Recommended Temperature: 4 - 60 °C. Reduce flow rate when
operating at low temperatures (e.g., 4 °C) to avoid excessive column
pressure.
7. Recommended Cleaning Solvents/Procedures:
A) 0.1 - 0.5M NaOH in HPLC grade water and if not successful,
B) 20 - 40% acetic acid in HPLC grade water or,
C) 30% acetonitrile or 30% methanol in HPLC grade water or,
D) 30% ethanol containing 0.5M NaOH or,
E) 8M Urea, 6M Guanidine or other nonionic surfactant HPLC
grade water
Clean the column regularly by injecting up to one column volume of
solution (A) (i.e., 0.1 - 0.5M NaOH in HPLC grade water) in 100 to
250 microliter increments. To maximize cleaning effect, reverse the
direction of eluent flow through the column and reduce the flow rate by
25%.
If desired, the HPLC column can also be cleaned by delivering, in
reverse eluent flow direction and at a 25 % standard flow rate, one of
the alternative cleaning solvents (i.e., 20 - 40% acetic acid in HPLC
grade water, or 30% acetonitrile or methanol in HPLC grade water, or
30% ethanol containing 0.5M NaOH, or either 8M Urea, 6M Guanidine
or other nonionic surfactant in HPLC grade water).
8. Recommended Storage: For overnight storage, continuously flush the
column with the mobile phase at 10 - 20% of the maximum recom-
mended flow rate. Store the column in the HPLC grade water when
it will be used within 24 hrs or in 20% ethanol containing 20mM
Tris-HCl buffer, pH 8 for long term storage.
VII. Waters BIosuIte deae and sP nP columns
This section contains recommended operating conditions and specifications
for the Waters non-porous, polymer-based, anion-and cation-exchange col-
umns.
Operating Conditions:
1. Shipping Solvent: The columns are shipped in HPLC grade water. Upon
receiving, flush the column with 30% acetonitrile/70% HPLC grade
water for 15 minutes at 1 mL/min. This is followed by equilibrating the
columns with the starting buffer.
Use high quality reagents, HPLC grade water, and HPLC grade solvents
for preparing buffers to maximize Waters BioSuite non-porous (NP)
column efficiency and life. Filter all eluents through a 0.20 or 0.45
µm membrane before use (Consult the with filtration membrane
manufacturer for details on solvent compatibility.) Fouling of the
non-porous resin, leading to a loss in retention and/or efficiency,
occurs faster compared to use of porous packing materials due to the
small surface area of non-porous resin particles. The useful column
lifetime is a function of numerous factors including the cleanliness and
composition of the mobile phase and the sample, the flow
rate and operating temperature, and the operating pressure. While
column cleaning is possible (see below), cleaning is not effective when
the column is damaged by irreversible sample
adsorption, channeling, or exposure of the packing material to exces-
sive heat of shock.
2. Recommended Operating Flow Rates/Max Pressure:
Note: When using a high viscosity buffer and ambient temperature, the maxi-
mum flow rate may have to be reduced to avoid exceeding the maximum
recommended operating pressure for the column. When changing solvents,
use 50% of the maximum flow rate.
3. Recommended pH Range: 2 - 12 (Avoid extended operation at pH
values less than 3 or greater than 10 to maximize column life.)
4. Recommended Salt Conc.: Less than or equal to 1.0 Molar
5. Recommended Organic Conc.: 0 - 20%.
Description Matrix i.d.
(mm)
L
(mm)
Flow Rates /
Max Pressure
BioSuite DEAE 2.5 µm
NP AXC polymer 4.6 35 1.0 - 1.6 mL/min /
3000 PSI
BioSuite SP 2.5 µm NP
CXC polymer 4.6 35 1.0 - 1.6 mL/min /
3000 PSI