BioHPLC Column Selection Guide Cover - Agilent Technologies

Microbore HPLC for sensitive peptide analysis
Column: PLRP-S 100Å 5 µm, 150 mm x various id
Mobile Phase: A: 0.01 M Tris HCI, pH 8
Flow Rate: 1 mL/min
B: A + 0.35 M NaCl, pH 8
Gradient: Linear 20% ACN, 0.1% TFA to 50% ACN,
0.1% TFA over 15 min
Injection Volume: 0.5 µL
Sample Conc: 0.25 mg/mL
Peptide separation on Agilent PLRP-S 100Å 5 µm columns
Peak Area
Peak Identification
1.0 mm
2.1 mm
4.6 mm
90
80
70
60
50
40
30
20
10
0
0 50 100 150
0
0 50 100 150
conc (ng)
conc (ng)
100
Standard curve data-point graphs on Agilent PLRP-S columns
80
60
40
20
MicroBore (1.0 mm id) Columns
1
A
B
C
0 min
8
Insulin 120 Oxytocin
160 Angiotensin II
90 Angiotensin I
Peak Area
Peak Area
Peak Identification
A. 1.0 mm id (flow rate 47 µL/min)
B. 2.1 mm id (flow rate 200 µL/min)
C. 4.6 mm id (flow rate 1 mL/min)
1. Oxytocin
2. Angiotensin II
3. Angiotensin I
4. Insulin
140
120
100
80
60
40
20
Peak Area
0
0
0 50 100 150 0 50 100 150
conc (ng)
conc (ng)
80
70
60
50
40
30
20
10
2
3
4
ANALYSIS SCALE
117