9.2
System Validation
(d)
Sample
*
Anthracene in acetonitrile
[Method for sample preparation]
Place 2OOmg of anthracene in the volumetric flask of
lOOmL capacity and add acetonitrile of HPLC grade
to it.
(e)
(f)
Isopropyl alcohol
Water (HPLC grade or equivalent)
(2)
(3)
Check the connection
of
the units. Refer to the instruction
manuals for details of the connection of each unit.
Before installing the column, observe tubing connection
of
the LC system. Use tubing with 0.3mm
I.D.
or
less
from
the
outlet of auto injector to the column inlet and
from
the column
outlet
to
the detector inlet. Length of the tubing should
be
30cm or less
to
minimize the extracolumn band broadening.
Flush the flow line with appropriate solvents dependent on
the operation status of the system. General guideline is
shown below.
To
flush the flow line with solvents, connect
the inlet and outlet tubing of the column directly using
a
proper
union.
Column should be connected after flushing.
(4)
0
Newly installed system
Flush
the flow line with isopropyl alcohol, then with water at
a flow rate of
2
mL/min
for ten minutes.
0
When
low
polarity solvent (such
as
hexane) is used
as
the mobile phase in the system
Flush the flow line with isopropyl alcohol, then with water at
a
flow rate of
2
Wmin for ten minutes.
0
When mixture of water and organic solvent, or
organic solvent miscible with water (methanol,
acetonitrile, etc.), or buffer solution is used as the
mobile phase in the system
Flush the flow line with water at a flow rate
of
2
mL/min
for
ten minutes.
(5)
Set the mobile phase (b) and flush the flow line, then connect
the column (c) to the LC system.
9
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10
-
RID
-
1OA
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