Waters XBridge Peptide BEH C18 User Manual

[ CARE AND USE MANUAL ]
1
CONTENTS
I. GETTING STARTED
a. Column Installation
b. Column Equilibration
c. Initial Column Efficiency Determination
II. COLUMN USE
a. Sample Preparation
b. Operating pH Limits
c. Solvents
d. Pressure
e. 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 SYSTEM
a. Column Connectors and
System Tubing Considerations
b. Measuring System Band Spreading Volume
c. Measuring Gradient Delay Volume
(or Dwell Volume)
VII. eCORD INTELLIGENT CHIP TECHNOLOGY
(applies only to XBridge XP 2.5 m,
≤4.6 mm I.D. Columns )
Thank you for choosing a Waters® XBridge® Peptide BEH
C
18
, 130Å or 300Å Column. The XBridge Peptide BEH C
18
,
130Å and 300Å packing materials were designed to provide
excellent peak shape, high efficiency, and excellent stability.
The XBridge Peptide BEH C
18
, 130Å and 300Å packing materials
are manufactured in a cGMP, ISO 9002 certified plant using
ultra pure reagent. Each batch of XBridge Peptide BEH C
18
Column material has been qualified with a peptide separation,
and the results are held to narrow specification ranges to
assure excellent, reproducible performance for peptide
separations. Every column is tested and a Performance Test
Chromatogram, along with a Certification of Acceptance, are
provided with each column.
XBridge Peptide BEH C
18
, 130Å and 300Å Columns
VIII. ADDITIONAL INFORMATION
a. Use of Narrow-Bore (3.0 mm I.D.)
b. Impact of Band Spreading Volume
on 2.1 mm I.D. Column Performance
c. Non-Optimized vs. Optimized
LC-MS/MS System:
System Modification Recommendations
IX. REPRESENTATIVE TEST CHROMATOGRAPH
AND CONDITIONS FOR SEPARATION OF
PROTEIN DIGEST
X. CAUTIONARY NOTE

Waters XBridge Peptide BEH C18 Specifications

General

Brand	Waters
Model	XBridge Peptide BEH C18
Category	Medical Equipment
Language	English

Summary

1. GETTING STARTED

1.a. Column Installation

Procedure for column installation, noting flow direction and detector connection.

1.b. Column Equilibration

Procedure for equilibrating the column with the desired mobile phase before use.

1.c. Initial Column Efficiency Determination

Performing an efficiency test upon receipt and at intervals for performance tracking.

II. COLUMN USE

2.a. Sample preparation

Guidelines for preparing samples, including filtration and solvent miscibility for optimal results.

2.b. Operating pH limits

Recommended pH range for columns and factors affecting column lifetime.

III. SCALING UP/DOWN ISOCRATIC METHODS

IV. TROUBLESHOOTING

V. COLUMN CLEANING, REGENERATION, AND STORAGE

5.a. Cleaning and Regeneration

Procedures for cleaning and regenerating columns due to contamination or performance issues.

5.b. Storage

Recommendations for storing columns to maintain performance and prevent bacterial growth.

VI. CONNECTING THE COLUMN TO THE HPLC SYSTEM

6.a. Column Connectors and System Tubing Considerations

Guidance on proper connection of tubing and fittings for optimal chromatographic results.

6.b. Measuring System Band Spreading Volume and System Variance

Procedure to measure system band spreading volume for HPLC system evaluation.

6.c. Measuring Gradient Delay Volume (or Dwell Volume)

Method for determining gradient delay volume for successful gradient method transfers.

VII. eCORD INTELLIGENT CHIP TECHNOLOGY

7.a. Introduction

Introduction to eCord Intelligent Chip for tracking column performance history.

7.b. Installation

Procedure for installing the eCord Intelligent Chip into the column heater.

7.c. Manufacturing Information

Overview of bulk material QC test results provided by the eCord Chip.

7.d. Customer Use Information

Details on column usage data captured by the eCord Chip, including injections and temperatures.

VIII. ADDITIONAL INFORMATION

8.a. Use of Narrow-Bore Columns

Guidelines for minimizing extra column effects and maximizing performance of narrow-bore columns.

8.b. Impact of Band Spreading Volume on 2.1 mm I.D. Column Performance

Explanation of how system band spreading affects performance in 2.1 mm I.D. columns.

8.c. Non-Optimized vs. Optimized LC-MS/MS System: System Modification Recommendations

Recommendations for system modifications to optimize LC-MS/MS performance with 2.1 mm columns.

IX. REPRESENTATIVE TEST CHROMATOGRAPH AND CONDITIONS FOR SEPARATION OF PROTEIN DIGEST

X. CAUTIONARY NOTE