Waters BioSuite Peptide Analysis Columns User Manual

Care and use manual, Biosuite peptide analysis columns

[ Care and Use ManUal ]

BioSuite Peptide Analysis Columns

Contents

I. IntroduCtIon

a. Column Performance Validation

b. Waters BioSuite C

3 μm PA-A and BioSuite C

3.5 μm

PA-B Columns

c. BioSuite C

3 μm, PA-A

d. BioSuite C

3 μm, PA-B

e. Operating Conditions

II. sample preparatIon

III. mobIle phase ConsIderatIons

IV. Flow dIreCtIon

V. ConneCtIng your Column to the hplC system—

Column ConneCtIon

VI. Column ConneCtors and system tubIng

ConsIderatIons

a. Column Performance Validation

VII. measurIng system bandspread Volume

VIII. measurIng gradIent delay Volume

IX. use oF bIosuIte “narrow-bore” peptIde analysIs

Columns (I.e., 2.1 mm I.d.)

X. system modIFICatIon reCommendatIons For bIosuIte
2.1 mm I.d. peptIde analysIs Columns

XI. addItIonal ConsIderatIons when usIng bIosuIte C

peptIde analysIs Columns

I. IntroduCtIon

Waters BioSuite

™

high performance chemistries are dedicated to the isola-

tion, analysis, and characterization of proteins and peptides. This Care
and Use document details the use of Waters BioSuite Peptide Analysis
Columns designed for peptide mapping applications. For the latest list
of BioSuite ion-exchange, size exclusion, hydrophobic interaction, and
reversed-phase columns, go to the “Life Sciences Chemistry Section” at
www.waters.com/lifesciences.

PLEASE READ CAREFULLY BEFORE USING THIS COLUMN

Thank you for choosing a Waters BioSuite Peptide Analysis Column. Every
batch of packing material is specifically tested using a tryptic digest of
cytochrome c. Only those batches that pass stringent tolerance values are
selected for use in BioSuite C

3 μm PA-A or C

3.5 μm PA-B Columns.

a. Column Performance Validation

Each BioSuite Peptide Analysis Column comes with a Certificate of Batch
Analysis and a Performance Test Chromatogram. The Certificates of Batch
Analysis are specific to each batch of packing material contained in the
BioSuite Peptide Analysis Column and includes the gel lot batch number,
analysis of unbonded particles, analysis of bonded particles, and chromato-
graphic results and conditions. The Performance Test Chromatogram is spe-
cific to each individual column and contains information such as gel lot batch
number, column serial number, USP plate count, USP tailing factor, capacity
factor and chromatographic results and conditions. These data should be
stored for future reference.

b. Waters BioSuite C

3 μm PA-A and BioSuite C

3.5 μm

PA-B Columns

BioSuite PA Series consists of two of Waters premier reversed-phase column
chemistries specifically voptimized for peptide mapping from simple to com-
plicated digests.

c. BioSuite C

3 μm, PA-A

BioSuite C

3 μm PA-A is a 100Å, diffunctional bonded, low ligand

density, silica-based column.

•

Specifically designed for excellent retention of polar peptides

• Ideal choice for LC/MS applications using formic acid (FA) that

minimizes ion-suppression

• Excellent performance for traditional HPLC-separations using low

TFA concentrations (e.g., 0.025% TFA)

bIosuIte peptIde analysIs Columns