Care and use manual – Waters BioSuite Columns User Manual
Page 5
[ Care and Use ManUal ]
BioSuite Columns
5
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: 2.5 - 7.5
4. Recommended Salt Conc.: Less than or equal to 0.5 Molar
5. Recommended Organic Conc.: 0 - 100% (Caution: Many proteins and
peptides are insoluble at elevated organic 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: 10-30 °C. Reduce flow rate when
operating at low temperatures (e.g., 10 °C) to avoid excessive column
pressure.
7. Recommended Cleaning Solvents/Procedures:
A) A concentrated salt solution at low pH, e.g. 0.5M Na
2
SO
4
,
pH 2.7
B) A low concentration of methanol or acetonitrile (e.g., 20%) in
HPLC grade water
C) A buffered solution of SDS, urea, or guanidine
Note: Choose a cleaning solvent based on sample properties, e.g. use (A) to
remove basic proteins, and (B) to remove hydrophobic proteins. Chaotrophic
agents can solvate strongly adsorbed proteins via hydrogen bond disruption.
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 either 20% ethanol or methanol
for long term storage. (Caution: Rinse column in HPLC grade water
before switching to 20% organic solvent to avoid salt precipitation in
column.)
9. Recommended Column Protection: Use of guard columns is recom-
mended to prolong the life of the analytical column life. Guard column
life depends greatly on sample cleanliness. As a general rule, guard
columns should be replaced after every 30-40 sample injections,
when the peaks become excessively wide, or when the peaks show
splitting.
VI. Waters BIosuIte Q-PeeK and sP-PeeK columns
This section contains recommended operating conditions and specifications
for the Waters polymer-based, anion and cation-exchange materials con-
tained in PEEK
™
columns.
Note: Compared to use of stainless-steel columns, connect the PEEK column
to the HPLC or Liquid Chromatographic System with a 10-32 polymer nut
and ferrule.
Operating Conditions:
1. Shipping Solvent: 20% Ethanol containing 20 mM Tris-HCl buffer
(pH 8.0)
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 2.5 Molar
5. Recommended Organic Conc.: 0 - 30%. (Caution: Some large
molecular weight proteins may be insoluble at elevated organic
Description
Matrix
i.d.
(mm)
L
(mm)
Flow Rates /
Max Pressure
BioSuite 250, 13 µm
SEC
silica
21.5
300
3.0 - 8.0 mL/min /
225 PSI
BioSuite 450, 13 µm
SEC
silica
7.5
300
0.5 - 1.2 mL/min /
225 PSI
BioSuite 450., 17 µm
SEC
silica
21.5
300
3.0 - 8.0 mL/min /
300 PSI
BioSuite Guard SEC
silica
7.5
75
0.5 - 1.2 mL/min /
600 PSI
BioSuite Guard SEC
silica
21.5
75
3.0 - 8.0 mL/min /
450 PSI
Description
Matrix
i.d.
(mm)
L
(mm)
Flow Rates /
Max Pressure
BioSuite Q-PEEK 10 µm
AXC
polymer
4.6
50
0.3 - 1.2 mL/min /
375 PSI
BioSuite SP-PEEK 7 µm
CXC
polymer
4.6
50
0.3 - 1.0 mL/min /
375 PSI