User guide

ManualsBrandsWaters ManualsWater equipmentPeptide Separation Technology ACQUITY UPLC BEH130 and BEH300

[ CARE AND USE MANUAL ]

CONTENTS

I. INTRODUCTION

II. GETTING STARTED

a. Column Connectors

b. Column Installation

c. Column Equilibration

d. eCord Installation

e. Initial Column Efficiency Determination

III. COLUMN USE

a. Sample Preparation

b. pH Range

c. Solvents

d. Pressure

e. Temperature

IV. COLUMN CLEANING, REGENERATING

AND STORAGE

a. Cleaning and Regeneration

b. Storage

V. INTRODUCING eCORD INTELLIGENT

CHIP TECHNOLOGY

VI. REPRESENTATIVE TEST CHROMATOGRAPH

AND CONDITIONS FOR SEPARATION OF

PROTEIN DIGEST

ACQUITY UPLC Peptide BEH C

, 130Å and 300Å Columns

I. INTRODUCTION

Thank you for choosing an ACQUITY UPLC Peptide BEH C

, 130Å or

300Å Column. The ACQUITY UPLC Peptide BEH C

, 130Å and 300Å

packing materials were designed specifically for use with the Waters

ACQUITY UPLC System and are manufactured in a cGMP, ISO 9002

certified plant using ultra pure reagents. Each batch of ACQUITY

UPLC BEH material is tested chromatographically with acidic, basic

and neutral analytes and has been qualitifed for peptide mapping.

The results are held to narrow specification ranges to assure

excellent, reproducible performance. Every column is individually

tested and a Performance Chromatogram and Certificate of Batch

Analysis are provided on the eCord™ intelligent chip. Each batch of

ACQUITY UPLC Peptide BEH C

, 130Å and 300Å Columns is also QC

tested with the separation of a protein digest.

Summary of content (6 pages)

PAGE 1
[ CARE AND USE MANUAL ] ACQUITY UPLC Peptide BEH C18, 130Å and 300Å Columns CONT ENT S I. INT RODUC T ION II. G E T T ING STA RT E D a. Column Connectors b. Column Installation c. Column Equilibration d. eCord Installation e. Initial Column Efficiency Determination III. COLUMN US E a. Sample Preparation b. pH Range c. Solvents d. Pressure e. Temperature IV. COLUMN C L EANING, R EG EN E RAT ING AND STO RAG E a. Cleaning and Regeneration b. Storage V.
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[ CARE AND USE MANUAL ] II. G E T T ING STA RT E D Note: If mobile phase additives are present in low concentrations (e.g., Each Peptide Separation Technology ACQUITY UPLC Peptide BEH ion-pairing reagents), 100 to 200 column volumes may be required C18, 130Å and 300Å Columns comes with Certificate of Analysis for complete equilibration. In addition, mobile phases that contain and a Performance Test Chromatogram embedded within the eCord formate (e.g., ammonium formate, formic acid, etc.
PAGE 3
[ CARE AND USE MANUAL ] III. COLUMN US E To ensure the continued high performance of ACQUITY UPLC Peptide questions. Alternatively, centrifugation for 20 minutes at BEH C18, 130Å and 300Å Columns follow these guidelines: 8,000 rpm, followed by the transfer of the supernatant liquid to an appropriate vial, could be considered. a. Sample Preparation b. pH Range 1. Sample must be dissolved in a diluent compatiable with initial strength of mobile phase.
PAGE 4
[ CARE AND USE MANUAL ] c. Solvents To maintain maximum column performance, use high quality chromatography grade solvents. Filter all aqueous buffers prior to use. Pall Gelman Laboratory Acrodisc ® filters are recommended. Solvents containing suspended particulate materials will generally clog the outside surface of the inlet distribution frit of the column. This will result in higher operating pressure and poorer performance. Table 3. Column Cleaning Sequence Polar Samples Proteinaceous Samples 1.
PAGE 5
[ CARE AND USE MANUAL ] At the time of manufacture, tracking and quality control information will be downloaded to the eCord. Storing this information on the chip will eliminate the need for a paper Certificate of Analysis. Once the user installs the column, the software will automatically download key parameters into a column history file stored on the chip. The eCord provides a solution to easily track the history of column usage.
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[ CARE AND USE MANUAL ] V I. R E P R E S ENTAT IV E T E ST C H ROMATOG RA P H AND CONDIT IONS FO R S E PA RAT ION / OF P ROT EIN DIG E ST T5 T12 T12-T13 T4 T9-T10 T8 T10 T14 T15 T1 T19 T13-T14 T19C 6.00 9.00 12.00 15.00 18.00 21.00 24.