HPLC Analysis of Lysozyme in Cheese on Chromolith(R) High Resolution RP-18e

HPLC Analysis of Lysozyme in Cheese on Chromolith® High Resolution RP-18e

Conditions

sample preparation 2g of homogenized hard cheese, add 20mL 1M NaCl solution in 100mL beaker and adjust to pH=6 with 0.1M NaOH,stir for 5min at 1200rpm, transfer the solution to another 100mL beaker and rinse old beaker with 10mL 1M NaCl solution. Stir one hour on a magnetc stirrer. (Adjust to pH=4.3 with 0.1M HCL and transfer the solution to 50mL flask, rinse the beaker with 1M NaCl into the flask and dilute to the mark with 1M NaCl. Filter the solution throuugh a folded filter and then through a 0.2μm PTFE syringe filter directly into a HPLC vial.)
column Chromolith® High Resolution RP-18 endcapped, 100 x 4.6 mm (1.52022.0001)
column temp. ambient
mobile phase [A] acetonitrile; [B] water + 0,1% TFA v/v;
gradient Initial composition: 0-20min 72,3% B, 20-21min.from 72,3% to 36,1%B, hold 36,1%B until 22min., 22-23.min from 36,1% to 72,3%B, hold 72,3%B until 35min.
flow rate 1.0 mL/min
sample Lysozyme 8.48mg/L; 2g hard cheese (Diluent: 1M NaCl)
injection 1 μL
detector 214 nm 1 μL

Description

Analysis Note Sample prep: Combine 2 g of homogenized hard cheese and 20 mL 1M NaCl solution in a 100 mL beaker. Adjust to pH 6 with 0.1M sodium hydroxide. Stir for 5 min at 1200 rpm. Transfer the solution to another 100 mL beaker and rinse old beaker with 10 mL 1M NaCl solution. Stir one hour on a magnetic stirrer. Adjust to pH 4.3 with 0.1M HCl and transfer the solution to a 50 mL flask, rinse the beaker with 1M NaCl into the flask and dilute to the mark with 1M NaCl. Filter the solution through a folded filter and then through a 0.2 μm PTFE syringe filter directly into an HPLC vial.
Featured Industry Food and Beverages
Legal Information Chromolith is a registered trademark of Merck KGaA, Darmstadt, Germany
suitability application for HPLC

Materials

     
Related Links