Biochromatography Applications

HPLC

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Biochromatography Applications

Trypsin digest of Lysosym 25 µg	Competitor n°1 Competitor n°2 Competitor n°3
Human Insulin	Competitor Uptisphere 120 Å 5µ C18-HDO, 250 x 4.6 mm
Breaking of peptide disulfide bridge	-
Proteins (MS/MS)	Uptisphere 300 Å 5µ C8, 100 x 0.125 mm

Trypsin digest of Lysosym 2S5 µg
Competitor n°1
	A : (H2O + 0.05% TFA) B : IPr 40 %+ (H2O + 0.05 % TFA) 60% Gradient : 0% —> 40% B in 30 min Detection UV UP5C4-25QS Courtesy of M. Malingue, Sedac Therapeutics
Competitor n°2

Competitor n°3

Human Insulin
Competitor	Uptisphere 120 Å 5µ C18-HDO, 250 x 4.6 mm

UP5HDO-25QS ACN 32% (H2O + 0.1 % TFA) 68% 1 ml/min UV 214 nm UV 276 nm

Breaking of peptide disulfide bridge

Competitor

TEAP Buffer at pH : 2.25

ACN

Gradient mode

1 ml/min

UV 220 nm

Limit of detection : 12.5 ng inj.

UP5ODB*25QS

Courtesy of Professeur Barthomeuf

Proteins (MS/MS)
Uptisphere 300 Å 5µ C8, 100 x 0.125 mm
UP5WC8Y10

A : 0.4% acetic acid + 0.005% HFBA B : 0.4% acetic acid Flow rate : 90 ml/min Split ~ 4µl/min Detection MS/MS

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