Waters ACQUITY APC Columns - Manual (Page 3)

3
ACQUITY APC Columns
Figure 1. Proper Tubing Column Connection.
Figure 2. Parker Ferrule in a Waters Style Endfitting.
d. eCord Installation
Attach the eCord button for each column to the side of the column
heater module noting their sequence. Up to four columns can
be installed at one time. The eCord button is magnetized and
does not require specific orientation. Once connected, column
identification and usage will be available though the instrument’s
control software.
e. Column Equilibration
ACQUITY APC Columns are shipped dry giving the user full
option of the solvent needed for the separation. Equilibrate the
column with a minimum of 20-column volumes, or until a stable
detector baseline is achieved. For a column bank, use the sum of
the column volumes in series to determine the total equilibration
volume. Refer to Table 2 for a listing of column volumes.
Table 2. Empty Column Volumes in mL
Column Length
(mm)
Column Internal Diameter
(mm)
Volume
(mL)
30 4.6 0.50
50 4.6 0.83
75 4.6 1.25
150 4.6 2.50
The rigid hybrid particle bed used for the ACQUITY APC Columns
allows the user to rapidly switch solvents without damaging the
column packing material. Changing solvents works best between
compatible solvents. For example, when changing between two
immiscible solvents, an intermediate solvent/co-solvent that is
miscible in both initial and final conditions should be used. For highly
viscous solvents, reduce the flow rate to avoid over pressuring the
system. Once the exchange is complete, equilibrate the column using
the final solvent conditions with a minimum of 20-column empty
volumes, or until a stable detector baseline is achieved.
III. COLUMN USE
a. Guidelines
ACQUITY APC Columns have a finite lifetime directly related to
their care and use. Column lifetime is reduced by contamination
from samples and eluents; improper handling and storage; and
exceeding operational conditions. To maximize ACQUITY APC
Column lifetime, pay attention to these guidelines:
■ ■
For best resolution and maximum column life, do not exceed
the flow rate recommendations found in Section III c.
■ ■
Protect the column from vibration and mechanical shock.
■ ■
Avoid precipitation by dissolving samples in the mobile
phase. If the sample is not dissolved in the mobile phase,
ensure that the sample, solvent and mobile phase are
miscible to avoid precipitation.
■ ■
Always use high-quality, particle-free, HPLC-grade solvents.
■ ■
Dedicate, whenever possible, the column to specific
applications. Frequent switching of samples and solvents
accelerates column deterioration and loss of resolution.
■ ■
Exceeding the upper temperature limit of the column.
■ ■
For aqueous mobile phases, take steps to avoid bacterial
contamination. For additional information refer to "Controlling
Contamination in UPLC
® /MS and HPLC/MS Systems", Waters
part number 715001307.
The presence of a void in the flow stream reduces column
performance. This can occur if a Parker ferrule is connected
to a Waters style endfitting (Figure 2).
In a proper tubing/column connection (Figure 1), the tubing touches
the bottom of the column endfitting, with no void between them.