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Title: PROBLEMS IN CEREAL PROTEIN ANALYSIS BY RP- AND SE-HPLC

Author
item Huebner, Floyd
item Bietz, Jerold

Submitted to: American Association of Cereal Chemists Meetings
Publication Type: Abstract Only
Publication Acceptance Date: 10/12/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Reversed-phase (RP-) and size-exclusion high-performance liquid chromatography (SE-HPLC) have become important methods for rapid identification of cereal varieties and for revealing quality differences. Accuracy and reproducibility are essential for good results. Recent studies have, however, indicated a possible problem in this area. Upon analysis of wheat, considerable protein, especially glutenin, may adhere t steel parts of the HPLC system and to the plastic injector rotor seal. Additional protein may be recovered after changing the rotor seal to Tefzel and the steel tubing to plastic (PEEK), including the sample loop. Others have reported that use of titanium or plastic instead of stainless steel in HPLC systems may reduce losses of enzymes; it is now apparent that cereal storage proteins may present similar problems. We have also found that addition of salt (0.1M guanidine hydrochloride) to storage protein samples -- even those extracted with 70% ethanol -- may improve protein recovery upon HPLC. Recognizing and taking the proper steps to deal with this problem will enhance the accuracy, reproducibility, and usefulness of HPLC for varietal identification and quality prediction.