Influence of hydrolysis, purification, and calibration method on furosine determination using Ion-pair reversed-phase high-performance liquid chromatography.
The influence of HCI concentration (6M, 8M, and 10M) and the ratio of sample protein to acid (1 or 5 mg of protein per mL of acid) on furosine formation during sample hydrolysis is studied. The conditions that maximize furosine formation are 10M HCI in the ratio of 1 mg of protein to 1 mL of acid. Purification of the hydrolysate by solid-phase extraction is also considered by examining the effect of hydrolysate volume and volume of 3M HCI used to elute the furosine. Furosine quantitation is carried out using the standard additions and external standard methods. The results indicate that there is no interference by the sample matrix and that external calibration is adequate.
Serrano
M A
MA
Departamento de Nutrición y Bromatología, Campus Universitario, Universidad de Alcalá, Alcalá de Henares (Madrid), Spain.
Castillo
G
G
Muñoz
M M
MM
Hernández
A
A
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006868:Hydrolysis; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D012015:Reference Standards
C018948:furosine; D008239:Lysine