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ACQUITY UPLC CSH130 C
18
Columns 7
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
2345678910 11 12 Min.
V. REPRESENTATIVE TEST CHROMATOGRAPH 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 6.71
Retention Time Difference (min) Result
T4 and T14 0.30
T10 and T9-T10 0.77
T19 and T19C 0.47
T12 and T12-T13 0.28
Chromatographic Conditions 
Column: 2.1 mm x 50 mm
Mobile Phase A: 0.1% formic acid in 100% water
Mobile Phase B: 0.085% formic acid in 75% acetonitrile reconstituted
w/ 200 µL of MPA, 5.0 µL injection
Sample: Tryptic digest of bovine cytochrome c (P/N 186006371)
Flow rate: 0.2 mL/min
Gradient: 1–21% B in 5 min; 21–31.4% B in 2.5 min; 31.4–95% B
in 2.5 min; 95% B for 1.5 min; 1%B for 2.5 min
Temperature: 40 °C Detection: UV 214 nm