HPLC Analysis of Proteins on BIOshell A400 Protein C18

column

BIOShell A400 C18, 10 cm x 2.1 mm I.D., 3.4 μm particles (67463-U)

mobile phase

[A] (water, 0.1% TFA); (acetonitrile, 0.08% TFA), 75:25; [B] (water, 0.1% TFA); (acetonitrile, 0.08% TFA), 25:75

gradient

20 to 100% B in 8 min

flow rate

0.3 mL/min

pressure

2480 psi total pressure (193 bar)

column temp.

30 °C

detector

UV, 215 nm

injection

2 μL

sample

lysozyme, 0.06; haptoglobulin, 0.06; conalbumin, 0.12; tryspin inhibitor, 0.18; glutamate dehydrogenase, (0.18; ß-galactosidase, 0.12; serum albumin chicken, 0.12 g/L in mobile phase A)