Care and use manual – Waters Oligonucleotide Separation Technology XBridge OST C18 Columns User Manual

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[ CARE AND USE MANUAL ]

I. GET TING STARTED

Each XBridge OST C

column is shipped with a Certificate of

Analysis and a Performance Test Chromatogram document. The
Certificate of Analysis is specific to the batch of packing material
contained in the XBridge OST C

column and includes the gel batch

number, analysis of unbonded particles, analysis of bonded particles,
and chromatographic results and conditions. The Performance Test
Chromatogram is specific to each individual column and contains
such information as gel batch number, column serial number, USP
plate count, USP tailing factor, capacity factor, and chromatographic
conditions. These data should be stored for future reference.

a. Connecting the Column

1. Purge the pumping system of any buffer-containing mobile phases

and connect the inlet end of the column to the injector outlet.

2. The XBridge OST C

column should be connected to HPLC with

1/16 inch tubing (stainless steel or PEEK

™

). An arrow on the column

identification label indicates the correct direction for solvent
flow. Make sure that XBridge OST C

column connection is made

with the correct ferrule seating adjustment. Figure 1 shows
the differences between Waters and Parker style of ferules and
end fittings. Note the differences in end fitting style and length of
tubing protruding from the ferrule end.

Figure 1: Waters and Parker Ferrule Types

Due to the absence of an industry standard, various column
manufacturers have employed different types of chromatographic
column connectors. The chromatographic 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. The
XBridge OST C

column is equipped with Waters style endfittings

that require a 0.130 inch ferrule depth. If a non-Waters style column
is presently being used, cut the end of the tubing with the ferrule,
place a new ferrule on the tubing and make a new connection. Before
tightening the screw, make sure that the tubing bottoms out in the
endfitting of the XBridge OST C

column.

In a proper tubing/column connection (Figure 2), the tubing touches
the bottom of the column endfitting, with no void between them.

Figure 2: Proper Tubing/Column Connection

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 3).

Note: A void appears if tubing with a Parker ferrule is connected to a
Waters style column.

Figure 3: Parker Ferrule in a Waters Style Endfitting

There is only one way 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 tightening

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).

Note: The connection leaks if a Waters ferrule is connected to a column
with a Parker style endfitting.

Waters Ferrule Setting

0.130 inches

Parker Ferrule Setting

0.090 inches

.090”

.130”

Void