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Care and use manual – Waters XSelect HSS HPLC Columns User Manual

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[ Care and Use ManUal ]

XSelect HSS HPLC Columns

2

II. ConneCtInG tHe CoLUMn to tHe HPLC sYsteM

a. Column Connection
Handle the column with care. Do not drop or hit the column on a hard
surface as this may disturb the bed and affect its performance.

1. Correct connection of 1/16 inch outer diameter stainless steel

tubing leading to and from the column is essential for high-
quality chromatographic results.

2. An arrow on the column identification label indicates correct

direction of solvent flow.

3. When using standard stainless steel compression screw fittings,

it is important to ensure proper fit of the 1/16 inch outer diameter
stainless steel tubing. When tightening or loosening the compression
screw, place a 5/16 inch wrench on the compression screw and a
3/8 inch wrench on the hex head of the column endfitting.

Caution: If one of the wrenches is placed on the column flat during this
process, the endfitting will be loosened and leak. Under-tightening
compression screws or using worn ferrules can lead to solvent leaking.
Care should be taken to check all column connections for leaks to
avoid exposure to solvents and the hazards associated with such exposure
including risks to health and electrical connections.

4. If a leak occurs between the stainless steel compression screw

fitting and the column endfitting, a new compression screw fitting,
tubing and ferrule must be assembled.

It is important to realize that extra column peak broadening can
destroy a successful separation. The choice of appropriate column
connectors and system tubing is discussed in detail below.

b. Column Connectors and System Tubing Considerations

Due to the absence of an industry standard, various column manufacturers
have employed different types of chromatographic column connectors.
The chromatographic performance of the separation can be negatively
affected if the style of the column endfittings does not match the
existing tubing ferrule setting. This section explains the differences
between Waters style and Parker style ferrules and endfittings (Figure
1). Each endfitting style varies in the required length of the tubing
protruding from the ferrule. XSelect HSS HPLC columns are equipped
with Waters style endfittings, which require a 0.130 inch ferrule. If
a non-Waters style column is presently being used, it is critical that

ferrule depth be reset for optimal performance prior to installing
the XSelect HSS HPLC column. In a proper tubing/column connection
(Figure 2), the tubing touches the bottom of the column endfitting,
with no void between them.

Figure 1: Waters and Parker Ferrule Types

Figure 2: Proper Tubing/Column Connection

Tubing touches the bottom of the column endfitting, with no void
between them.

Attention: A void will occur if a Parker ferrule connection is used with
a Waters style endfitting (Figure 3). This will dramatically reduce the
efficiency of the column and cause peak shape distortion.

Figure 3: Parker Ferrule in a Waters Style Endfitting

To fix this problem: Cut the end of the tubing with the ferrule, place a
new ferrule on the tubing and make a new connection. Before tighten-
ing the screw, make sure that the tubing bottoms out in the endfitting
of the column.

Conversely, if tubing with a Waters ferrule is connected to a column
with Parker style endfitting, the end of the tubing will bottom out
before the ferrule reaches its proper sealing position. This will leave a
gap and create a leak (Figure 4).

Caution: The connection will leak if a Waters ferrule is connected to a
column with a Parker style endfitting.

Waters Ferrule Setting

Parker Ferrule Setting