SELECTION OF COLUMN AND OPERATING CONDITIONS FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PROTEINS IN CANADIAN WHEAT | Zendy

J. E. Kruger | Zendy; B. A. Marchylo | Zendy

Open Access

SELECTION OF COLUMN AND OPERATING CONDITIONS FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PROTEINS IN CANADIAN WHEAT

Author(s) -

J. E. Kruger,

B. A. Marchylo

Publication year - 1985

Publication title -

canadian journal of plant science

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.338

H-Index - 59

eISSN - 1918-1833

pISSN - 0008-4220

DOI - 10.4141/cjps85-041

Subject(s) - chromatography , high performance liquid chromatography , chemistry , elution , dithiothreitol , acetic acid , resolution (logic) , biochemistry , artificial intelligence , computer science , enzyme

Chromatographic conditions were optimized and three commercially available columns were evaluated for separation of alcohol-soluble storage proteins of Neepawa wheat using reversed-phase high-performance liquid chromatography (RP-HPLC). Optimal separation was achieved using an extracting solution of 50% 1-propanol, 1% acetic acid, and 4% dithiothreitol and an HPLC elution time of 105 min at a flow rate of 1.0 mL/min. HPLC columns evaluated (SynChropak RP-P, Ultrapore RPSC and Aquapore RP-300) varied in selectivity and resolution. The column providing the greatest versatility was Aquapore RP-300 available in cartridge form. Sodium dodecyl sulfate gradient-gel electrophoresis analysis of protein peaks resolved by RP-HPLC indicated that many of the eluted peaks contained more than one protein species. Chromatographic protein patterns obtained for Neepawa wheat grown at different locations and in different years were qualitatively the same.Key words: Protein, high-performance liquid chromatography, wheat

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