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Separation of tryptic digests by HPLC

Method: Matrix Application-No.:

HPLC - 108200

Substances: cytochrome C; bovine serum albumin (BSA)

Columns: A) EC 125/4 NUCLEOSIL® 100-5 C18 PPN

B) EC 250/4 NUCLEOSIL® 100-5 C18 PPN

Phase: NUCLEOSIL® C18 PPN

REF numbers: A) 720251.40

B) 720252.40

Conditions: Method A)
Eluent A: 0.1 % trifluoroacetic acid (TFA) in water
Eluent B: 0.08 % trifluoroacetic acid (TFA) in acetonitrile
Gradient: from 0 to 35 % B in 60 min
Flow rate: 1 mL/min
Injection volume: 10 µL

Method B)
Eluent A: 0.1 % trifluoroacetic acid (TFA) in water
Eluent B: 0.08 % trifluoroacetic acid (TFA) in acetonitrile
Gradient: from 0 to 40 % B in 90 min
Flow rate: 1 mL/min
Injection volume: 10 µL

Detection A) UV, 220 nm

Subsequent B) UV, 215 nm
analysis:

Author(-s): LC department

Source: MACHEREY-NAGEL

Keywords: proteins; tryptic digest; peptides

Chromatograms:
A) Separation of a tryptic digest of cytochrome C

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Separation of tryptic digests by HPLC https://www.mn-net.com/DesktopModules/TabID/9657/defauld.aspx

B) Separation of a tryptic digest of bovine serum albumin

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