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Waters XSelect CSH130 C18 3.5 μm and 5 μm Columns User Manual

Care and use manual, Xselect csh130 c, 5 µm and 5 µm columns

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[ CARE AND USE MANUAL ]

XSelect CSH130 C

18

Columns

1

Thank you for choosing a Waters XSelect

CSH

HPLC column. XSelect

CSH130 C

18

, 3.5 µm and 5 µm Columns feature Waters Charged Surface

Hybrid (CSH) Technology which provides excellent peak shape, high
efficiency and loading capacity for basic compounds when using acidic,
low ionic strength mobile phases. This same particle technology is used
in the ACQUITY UPLC CSH130 C

18

, 1.7 µm and 2.5 µm XP family of

HPLC columns, thus enabling seamless transferability between HPLC and
UPLC

®

system platforms. The XSelect CSH130 C

18

packing materials

were designed and are manufactured in a cGMP, ISO 9001 certified
manufacturing facility using ultra pure reagents. Each batch of XSelect
CSH130 C

18

material is tested chromatographically with acidic, basic

and neutral analytes as part of qualification for use in peptide mapping.
The results are held to narrow specification ranges to assure excellent,
reproducible performance. XSelect CSH130 C

18

batches are also QC tested

with a gradient separation of a tryptic digest of cytochrome c using 0.1%
formic acid containing eluents. Finally, every shipped column is individually
tested for packed bed efficiency and a Performance Chromatogram and
Certificate of Batch Analysis is available upon request.

CONTENTS

I. GETTING STARTED

a. Column Installation
b. Column Equilibration
c. Initial Column Efficiency Determination

II. COLUMN USE

a. Guard Columns
b. Sample Preparation
c. Operating pH Limits
d. Solvents
e. Pressure
f. Temperature

III. SCALING UP/DOWN ISOCRATIC METHODS

IV. TROUBLESHOOTING

V. COLUMN CLEANING, REGENERATING AND STORAGE

a. Cleaning and Regeneration
b. Storage

VI. CONNECTING THE COLUMN TO THE HPLC

a. Column Connectors and System Tubing Considerations
b. Measuring System Bandspreading Volume
c. Measuring Gradient Delay Volume (or Dwell Volume)

VII. ADDITIONAL INFORMATION

a. Use of Narrow-Bore (2.1 mm i.d.)
b. Impact of Bandspreading Volume on 2.1 mm i.d. Column Performance
c. Non-Optimized vs. Optimized LC/MS/MS System: System
Modification Recommendations

XSELECT CSH130 C

18

, 3.5 µm AND 5 µm COLUMNS