Teledyne ISCO RediSep Rf Gold User Manual

8.6 kg RediSep Rf Gold®

CV or Time

Reversed Phase C18 Columns

Use, Cleaning, and Storage

Chromatography Application Note AN116

#692203917, Rev , Feb 23, 2021

Application Overview

Proper sample preparation, care, cleaning, and storage
of reusable Teledyne ISCO RediSep Rf Gold high
performance C18 columns will help to extend column
life and improve column consistency and reliability.

General Method

Column Equilibration

The column comes preconditioned in a 1:1 mixture of
water:methanol. Before your first run or when changing
solvent systems, flush the column in manual control
with 3CV of the solvent composition of the method’s
starting gradient conditions.

For future runs in the same solvent system use the
equilibration period built into the default method.

Column Media Column Optimal Flow Rate
Weight Volume (mL) (mL/min)

RediSep Rf Gold High Performance C18 Columns

5.5 4.6 18

15.5 13.5 30

30 26.4 35

50 42.6 40

100 87.7 60

150 129.7 85

275 243.4 50

415 363.5 150

950 825 200

1900 1650 180

3800 3000 260

8600 7500 360

Table 1: Column Data

Sample Loading

The sample load on the column should be 0.1 to 2 percent
of the column media weight.

Proper sample preparation is critical to maximizing the
lifespan of the column. This means the sample should
be free of particulates and well soluble in the loading

solvent.

Post-run Care

With proper care, RediSep Rf Gold C18 columns may
be used thirty times or more. Reversed phase methods
should be written so that at the end of the separation,
the column is washed with 2–4CV of a high percentage
(100%) of organic solvent. After 2– 4CV of wash, the
percent organic can be returned to the starting condition
for 2–3CV to anticipate the next run column equilibration
conditions (see Figure 1). Alternatively, the percent
organic can be changed to 20–90% in anticipation of
column storage.

100

Column

End of

Separation

Wash

80 %B

for Column

Storage

OR

Equilibrate/

Next Initial %B

75

50

Separation

Gradient

25

Gradient % Solvent B

0

Figure 1: Post Run Column Flushing

Initial %B

Storage

It is possible to store C18 columns for long periods with
no loss in performance. Following are some general
rules for storage for reversed phase C18 columns:

•Once wetted, never dry out a reversed phase column.

This will induce channeling due to expansion and
contraction of the stationary phase. The air purge
feature (if available on the Flash chromatography
system) should be turned off.

•Store columns wet and well-capped in 10–80% water
and 20–90% organic solvent such as acetonitrile,
methanol, or ethanol.

•Do not store the column under acidic, basic, or
buffered conditions, be sure to flush the column

and store under the conditions noted above.

8.6 kg Column Installation Instructions

1. The 8.6 kg column is to large to fit on the Torrent,
enough tubing is supplied to place the column next
to the Torrent or on the floor if needed. The column is
shipped wet and weights over 55 pounds, two people
should help position the column when unpacking.

2. Remove the black plastic nut
from the top of the Torrent’s
column stand. Then take two
wrenches to remove the nut from
the adapter (circled in red).

3. Connect the tube connected to
the Torrent and the now free
nut and ferrule and connect to
the Union attached to the pre­swaged tubing.

4. Remove the swaged plug from
the top of the column with a
wrench and replace with the
tubing that is now connected
via union to the Torrent.

5. Remove the swaged plug from
the bottom of the column with a
wrench and replace it with the
second piece of tubing that came
with the column.

6. Remove the black fitting on the
bottom port of the Torrent and
connect the tubing from the
bottom of the column to this port
with the black nut.

7. Column is now ready to be used following the use
and care guidelines on the back of this sheet.

Summary

Clean columns with high (100%) organic solvent. Never
dry the column and store the column wet and capped
with 20–90% organic solvent.

Teledyne ISCO

P.O. Box 82531, Lincoln, Nebraska, 68501 USA
Toll-free: [phone] • Phone: [phone] • Fax: [phone]

Teledyne ISCO is continually improving its products and reserves the right to change product
specifications, replacement parts, schematics, and instructions without notice.

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