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Waters XBridge Protein BEH, C4, 300A, 3.5 µm Columns User Manual

Xbridge protein beh c, Care and use manual

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[ CARE AND USE MANUAL ]

XBridge Protein BEH C

4

, 300Å, 3.5, 5, and 10 µm Columns

I. INT RODUCTION

Thank you for choosing a Waters reversed-phase Protein Column.
The XBridge

®

Protein BEH C

4

, 300Å packing material was designed

to provide excellent peak shape, high efficiency, and good recovery
for biological macromolecules that are too large or too hydrophobic
for separation on columns with smaller pores or longer chain
bonded phases. The base particle and bonding chemistry are chosen
to provide exceptional stability at both high and low pH as well as
at high temperature. The XBridge Protein BEH C

4

, 300Å packing

material is manufactured in a cGMP, ISO9002-certified plant
using ultra-pure reagents. Each batch of XBridge Protein BEH C

4

reversed-phase column material has been qualified with a protein
test mixture, and the results are held to narrow specification ranges
to ensure reproducible performance. Every column is individually
tested for efficiency, and a Performance Test Chromatogram along
with a Certification of Acceptance is provided with each column.

CONTENTS

I. INT RODUCTION

II. GET TING START ED

a. Column Installation

b. Column Equilibration

c. Initial Column Efficiency Determination

d. Useful Functional Tests for Benchmarking

a New Column

III. COLUMN USE

a. Sample Preparation

b. Operating pH Limits

c. Solvents

d. Pressure

c. Temperature

IV. SCALING SEPARATIONS

V. T ROUBLESHOOTING

VI. COLUMN CLEANING, REGENERATING,

AND STORAGE

a. Cleaning and Regeneration

b. Storage

VII. CONNECTING T HE COLUMN TO T HE HPLC

a. Column Connectors and System

Tubing Considerations

b. Measuring System Bandspreading Volume

VIII. MEASURING GRADIENT SYST EM VOLUME

(OR DW ELL VOLUME)