HPLC Analysis of Human Serum Albumin Tryptic Digest on Ascentis® Express C18

HPLC Analysis of Human Serum Albumin Tryptic Digest on Ascentis® Express C18

Conditions

sample/matrix human serum albumin, tryptic digest
column Ascentis Express C18, 3 cm x 4.6 mm I.D., 2.7 µm particles (53818-U)
column temp. 45 C
mobile phase [A] 10 mM ammonium acetate in water, pH 9; [B] 10 mM ammonium acetate in methanol/water, pH 9; (90:10, A:B)
gradient 0% B in 0.50 min; 55% B in 0.25 min; 90% B for 0.50-0.69 min; 0% B for 0.70-1.00 min
flow rate 4.0 ml/min
injection 20 µl
detector UV, 254 nm

Description

Categories Analytical Chromatography, Proteins
Featured Industry Life Science and Biopharma
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
Other Notes Sample Prep: 0.1 gram of albumin was dissolved in 10 mL of 0.01 M ammonium formate buffer, and the pH was adjusted to 8.0 with ammonium hydroxide; the solution was heated in a boiling water bath for 6 min. After the solution cooled, 2.0 mg of trypsin from bovine pancreas was added, and the mixture was allowed to react for 4 h at 37 °C; the reaction was quenched by adding 0.1% trifluoroacetic acid to pH 2. The digest was stored at 4 °C and filtered prior to injection through a 0.45 ìm nylon membrane
suitability application for HPLC

Materials

     
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