Waters XSelect CSH130 C18 3.5 μm and 5 μm Columns - Manual (Page 10)

[ CARE AND USE MANUAL ]
XSelect CSH130 C
18
Columns 10
1.20
1.10
1.00
0.90
0.80
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
2 3 4 5 6 7 8 9 10 11 12 Min.
Figure 10: Representative Test Chromatography and Conditions for Separation of a Cytochrome c Tryptic Digest
Peak Identification
T1 N-AcGDVEK
T13-T14 KYIPGTK
T14 YIPGTK
T4 IFVQK
T9-T10 KTGQAPGFSYTDANK
T10 TGQAPGFSYTDANK
T8 TGPNLHGLFGR
T15 MIFAGIK
T19C EDLIAY
T19 EDLIAYLK
T12-T13 GITWGEETLMEYLENPKK
T12 GITWGEETLMEYLENPK
T5 CAQCHTVEK (heme attached)
Retention Time (min) Result
T19C 6.69
Retention Time Difference (min) Result
T4 and T14 0.312
T10 and T9-T10 0.780
T19 and T19C 0.461
T12 and T12-T13 0.290
Chromatographic Conditions
Column: 2.1 mm x 50 mm
Sample: Tryptic digest of bovine cytochrome c (P/N 186006371)
Reconstituted w/ 200µL of MP A, 5.0µL injection
Flow rate: 0.2 mL/min
Temperature: 40°C
Mobile Phase A: 0.1% formic acid in 100% water MPA
Mobile Phase B: 0.085% formic acid in 75% acetonitrile MPB
Gradient: 1 to 21%B in 5 min; 21 to 31.4% B in 2.5 min; 31.4 to
95% B in 2.5 min; 95% B for 1.5min; 1%B for 2.5 min
Detection: UV 214 nm