Determination of molecular-mass distribution of food-grade protein hydrolyzates by size-exclusion chromatography and chemiluminescent nitrogen detection.
Size-exclusion chromatography (SEC) and chemiluminescent nitrogen detection (CLND) were used to estimate the molecular-mass distribution of food-grade protein hydrolyzates. Simultaneous CLND and UV (214 nm) detection is demonstrated for analytical SEC of an experimental casein hydrolyzate. In order to validate the estimated average M(r) values derived from the SEC column data, a preparative SEC separation of an extensive casein hydrolyzate was pursued. Fractions were collected on a time basis and analyzed by time-of-flight (TOF) mass spectrometry. A plot of TOF M(r) vs. SEC M(r) indicated that the peptides below M(r) of 1200 were eluted as estimated by the calibrated preparative SEC column. This paper demonstrates the power of using a dual CLND and UV detection system for analytical SEC analysis of protein hydrolyzates with a calibrated column.
Fujinari
E M
EM
Antek Instruments Inc., Houston, TX 77090, USA.
Manes
J D
JD
D002364:Caseins / Q000737:chemistry; D002850:Chromatography, Gel; D005504:Food Analysis; D006868:Hydrolysis; D008163:Luminescent Measurements; D008970:Molecular Weight; D009584:Nitrogen / Q000032:analysis*; D011506:Proteins / Q000737:chemistry*
D002364:Caseins; D011506:Proteins; D009584:Nitrogen