Submitted to: Cereal Foods World
Publication Type: Peer reviewed journal
Publication Acceptance Date: 3/27/2003
Publication Date: 4/18/2003
Citation: Lookhart, G.L., Bean, S.R. and Bietz, J.A. 2003. Reversed-phase high performance liquid chromatography in grain applications. Feature Article. Cereal Foods World 48:9-17. Interpretive Summary: Reversed-phase high performance liquid chromatography (RP-HPLC) is an important tool for studying cereal proteins. RP-HPLC is fully automated and produces data that is easy to analyze and store, making RP-HPLC an important and efficient tool to study cereal proteins and their relationships to grain quality. Over the years many methods of RP-HPLC have been used to separate and study cereal proteins. This overview describes methods proven to work for analyzing cereal proteins and offers detailed protocols for sample preparation and analysis methodology. This information is important for anyone wishing to use RP-HPLC to study cereal proteins.
Technical Abstract: High-performance liquid chromatography (HPLC) uses a liquid pumping system to accurately deliver solvents through a column packed with 1.5- to 10-µm particles with specific bonded phases. The end result is the ability to separate complex mixtures in minutes. This article focuses on the separation of gluten monomers using reversed-phase HPLC (RP-HPLC). RP-HPLC is a method often used for gluten protein analysis. Its resolution equals or exceeds that of most other methods, and it is fast, reproducible, sensitive, quantifiable, and gives good recovery. Most importantly, however, it complements other methods, since it fractionates proteins on the basis of different surface hydrophobicities.