Waters BioSuite Columns - Manual (Page 3)

[ Care and Use ManUal ]
BioSuite Columns 3
the flow in this reversed flow direction until a few drops of mobile phase
exit from the column. Turn off the flow, let the pressure go to zero, and
disconnect the column from the system. Turn the column around and hook
it up so that the flow is now in the direction of the arrow. Start the flow
at a low setting and stop it as soon as the mobile phase exits from the
bottom fitting. Now you can hook up the column to the detector inlet and
increase the flow to the desired setting.)
• Be sure to set the flow within the recommended range shown in the
following tables.
Sample
If possible, always dissolve your sample in mobile phase or the starting
mobile phase when operating under gradient conditions. Alternatively, try
to match the pH, salt concentration and organic solvent of the sample with
those of the mobile phase and run a test to ensure that no precipitate, sus-
pension or flocculate is formed. Finally, before making an injection, filter the
sample through a 0.20-0.45 µm porosity membrane.
III. Waters BIosuIte Pc
18
and PPhenyl rPc columns
This section contains recommended operating conditions and specifications
for the Waters polymer-based, C
18
and phenyl reversed-phase columns.
Operating Conditions
1. Shipping Solvent: 70% Methanol - 30% Water
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 (20% for the 21.5 mm i.d. preparative
column).
3. Recommended pH Range: 2 - 12 (Avoid extended operation at pH >
10 to maximize column life.)
4. Recommended Organic Concentration Range: 0 - 100% (Note:
Reduced organic concentrations may be required if buffers are present
in the eluent in order to avoid salt precipitation.)
5. Recommended Temperature Range: 5 - 50 °C (Note: Reduce flow rate
when operating at low temperatures (e.g.. 5 C) to avoid excessive
column pressure.)
6. Recommended Cleaning Solvents/Procedures:
A) Acetonitrile or Methanol, or
B) Aqueous buffer in organic solvent, or
C) 0.1 - 0.2M NaOH, or
D) 20 - 40% acetic acid in HPLC grade water.
Clean the column regularly with any of the listed solvents. Cleaning
solvents (C) and (D) can be injected onto the column in 250 µl incre-
ments for a total of one column volume. Use proportionally larger
volumes for the 21.5 mm i.d. prep column. Following use of cleaning
solvent, rinse the column with 3 to 5 column volumes of HPLC grade
water before storage.
7. 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 shipping solvent when the
column will not be used within 48 hours. Description Matrix i.d.
(mm)
L
(mm)
Flow Rates / Max
Pressure
BioSuite pC
18
, 2.5 µm
NP RPC
polymer 4.6 35 1.0 - 1.6 mL/min /
3000 PSI
BioSuite pC
18
, 500,
7 µm RPC
polymer 4.6 150 0.08 - 0.22 mL/min
/ 1500 PSI
BioSuite pC
18
, 500,
7 µm RPC
polymer 4.6 150 0.5 - 1.2 mL/min /
1750 PSI
BioSuite pC
18
, 500,
13 µm RPC
polymer 21.5 150 3.0 - 8.0 mL/min /
500 PSI
BioSuite pPhenyl, 1000,
10 µm RPC polymer 2.0 75 0.05 – 0.12 mL/min
/ 150 PSI
BioSuite pPhenyl, 1000,
10 µm RPC polymer 4.6 75 0.5 - 1.2 mL/min /
450 PSI
BioSuite pPhenyl, 1000,
13 µm RPC polymer 21.5 150 6.0 - 8.0 mL/min /
450 PSI