BioHPLC Column Selection Guide Cover - Agilent Technologies

Capillary and Nano Columns
Capillary columns for HPLC analyses
with UV and MS detection
Column: ZORBAX 300SB-C18
5064-8263
0.3 x 150 mm, 5 µm
Mobile Phase: 5-55% B in 50 min, to 85% B from 55-57 min
A: Water + 0.1% Formic acid
B: Acetonitrile + 0.1% Formic acid
Flow Rate: 5.5 µL/min
Detector: 206 nm
MS Conditions: LC/MS: Pos. Ion ESI
with LC/MSD trap-Vcap 4000 V
Drying Gas Flow:
Drying Gas
7 L/min
Temperature: 250 °C
Nebulizer: 15 psi
Capillary Exit Volt: 50 V
Max Accum Time: 300 ms
Total Averages: 3
Isolation Width: 3 m/z
Frag Ampitude: 1.0 V
Sample: 100 nL
Beta Casein Digest (4 pmol)
A ZORBAX 300SB-C18 capillary column (0.3 mm id) is used
for the separation of the protein digest. Detection is by both
UV and Electrospray MS. MS detection can be used for
identification of peptide fragments.
110 www.agilent.com/chem/biohplc
UV at 206 nm
% Relative Abundance
100
80
60
40
20
0
x10
0.8
0.6
0.4
0.2
0.0
UV
MS
10
10
15
15
20 25 30 35 40 45 50
Time (min)
20 25 30 35 40 45 50
Time (min)
LCSB007