Owner's manual

ManualsBrandsWaters ManualsWater equipmentBioSuite Peptide Analysis Columns

[ Care and Use ManUal ]

BioSuite Peptide Analysis Columns 7

Note: Flow splitters after the column will introduce additional bandspread-

ing. System optimization, especially in a system that contains a flow split-

ter, can have dramatic effects on sensitivity and resolution. Optimization

includes using correct-depth ferrules and minimizing tubing diameter and

lengths. An example is given in Figure 9 where system optimization resulted

in a doubling of sensitivity and resolution of the metabolite in an LC/MS/MS

system.

Non-optimized LC/MS/MS System Optimized System

Figure 9: Chromatographic Performance on Non-Optimized vs. Optimized

LC/MS/MS System.

X. system modIFICatIon reCommendatIons For bIosuIte

2.1 mm I.d. peptIde analysIs Columns

1. UseamicroboredetectorflowcellwithBioSuite2.1mmi.d.Peptide

Analysis Column.

Note: Detector sensitivity is reduced with the shorter flowcell pathlength in

order to achieve lower bandspread volume.

2. Minimizeinjectorsampleloopvolume.

3. Use0.009inch(0.25mm)tubingbetweenpumpandinjector

4. Use0.009inch(0.25mm)tubingforrestofconnectionsinstan-

dardsystemsand0.005inch(0.12mm)tubingfornarrowbore

(>2.1mmi.d.)systems.

XI. addItIonal ConsIderatIons when usIng bIosuIte C

peptIde analysIs Columns

WatersBioSuitePeptideAnalysisColumnscontaineither3μm(i.e.,BioSuite

3μmPA-A)or3.5μm(i.e.,BioSuiteC

3.5μmPA-B)particlesthatpro-

vide very efficient separations without sacrificing column lifetime.

This section describes five parameters to consider when performing peptide

separations with BioSuite Peptide Analysis Columns.

Note: Columns that contain 3 μm or 3.5 μm particles have smaller outlet

frits to retain packing material. These columns should not be backflushed.

1. Backpressure: Backpressures for columns containing either 3 μm or

3.5 μm particles are higher than seen using columns packed with 5 μm

or 10 μm material.

2. Temperature:AsseeninTable1,useofacolumnheatersetat

elevatedtemperature(e.g.,40°C)willsignificantlyreducethe

observed system backpressure when using BioSuite Peptide Analysis

Columns.

3. Sampling Rate: Use a data sampling rate of 10 points per second or

greater.

4. DetectorTimeConstant:Useatimeconstantof0.1secondsforfast

analysis.

7.00 7.50

Non-optimized LC/MS/MS System Optimized System

8.00

7.00 7.50 8.00