Care and use manual – Waters Delta-Pak High Pressure Insert HPLC Column User Manual

Page 5

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[ Care and Use ManUal ]

5

Table 2: Viscosity

The typical operating backpressure for Waters

®

Delta-Pak

TM

columns

is summarized below.

Table 3: Typical Operating Backpressure at 1 mL/min

A variation of plus or minus 10-15% is acceptable.

d. Column Temperature Limits

The column hardware has been designed to operate between 4 °C and
50 °C.

Note: Column failure due to operation outside of these temperature
limits voids the warranty.

V. care and MaIntenance

a. Troubleshooting

Table 4, on the following page, provides the corrective action for
specific problems that may occur with the Waters HPLC High Pressure
Inert columns.

b. Cleaning the Column

Flush reversed-phase columns with methanol or acetonitrile. It is
useful to monitor the UV absorbance during this procedure to see if
contaminants are being removed from the column.

If flushing does not resolve the problem, wash the column with sol-
vents of decreasing polarity, always making sure the solvent in the
column and a washing solvent are miscible. Return the column to the
standard reversed-phase conditions by reversing the wash sequence.

If you suspect a particular contaminant is building up on the column,
use solvents or chemicals that are known to dissolve a material.
Be sure to remain within the limits of pH 2.0 to pH 8.0. Also,
ensure that no precipitate forms as a result of mixing the washing
solution with the solvent in the column. Proteinaceous material can
sometimes be removed by making repeated injections of 200 µL of
dimethylsulfoxide (DMSO).

c. Storing the Column

Leaving the column unused for less than 72 hours does not generally
require specialized storage procedures. However, never let columns
dry out. For longer storage:

• DO NOT store the column in water alone, as this may result

in bacterial growth in the column. Storing the column in a 10%
aqueous solution of isopropanol or methanol will hinder
bacterial growth.

• DO NOT store the column in buffered solution or solutions

containing salts, as these may precipitate.

• Return the column to its box with the end plugs firmly in place

for storage. Allowing the column to dry out may result in poor
chromatographic performance.

• DO NOT leave a column at elevated temperatures with no mobile

phase flow.

Column

Dimension Mpa Methanol

(Water)

Atm Metha-

nol (Water)

Psi Methanol

(Water)

Delta-

Pak

TM

2.1 x 150

mm

17 (28)

170 (285)

2500 (4200)

Delta-

Pak

TM

3.9 x 150

mm

6.0 (10.0)

60 (100)

900 (1500)

Mobile Phase

Viscosity (20 °C),
Centipoise

n-Pentane

0.24

n-Hexane

0.33

wn-Heptane

0.42

Isoctane

0.50

Acetone

0.32

Dioxane

1.54

Nitromethane

0.65

Acetonitrile

0.37

n-Propanol

2.3

Ethanol

1.2

Methanol

0.6

Water

1.0

60/40 Methanol/Water

2.0

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