Waters BioSuite Peptide Analysis Columns - Manual

[ Care and Use ManUal ]
BioSuite Peptide Analysis Columns 1
Contents
I. IntroduCtIon
a. Column Performance Validation
b. Waters BioSuite C
18
3 μm PA-A and BioSuite C
18
3.5 μm
PA-B Columns
c. BioSuite C
18
3 μm, PA-A
d. BioSuite C
18
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
18
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
18
3 μm PA-A or C
18
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
18
3 μm PA-A and BioSuite C
18
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
18
3 μm, PA-A
BioSuite C
18
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