Scientia Agricultura Sinica ›› 2007, Vol. 40 ›› Issue (5): 1002-1009 .doi: 10.3864/j.issn.0578-1752.at-2005-6305

• HORTICULTURE • Previous Articles     Next Articles

Protocal establishment of Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) for Analyzing Wheat Gluten Protein

  

  1. 华中农业大学植物科技学院
  • Received:2005-10-28 Revised:1900-01-01 Online:2007-05-10 Published:2007-05-10

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Abstract: Reversed-phase high-performance liquid chromatography (RP-HPLC) is an effective method in separating wheat gluten protein, which has been widely used in wheat quality study. Two cultivars, Zhongyou 9507 and Jing 411，were chosen to study the effect of temperature, elution gradient, sample weight, extraction duration, injection volume on gluten protein resolution and the repeat ability of these analyses. The results indicated that the gliadin and glutenin subunits extraction rate can reach 90% when 45mg of flour was extracted with 1ml 50% aqueous 1-propanol (v/v) and glutenin extraction buffer, respectively. The optimal sample injection volume of gliadin and glutenin was 10~15 ?l and 15~20 ?l, respectively. The modified elution gradient can separate all types of gliadin and glutenin subunits. This new protocol can qualify and quantify the components of gliadin and glutenin accurately, by which we can predicate the wheat end-use quality.

Key words: aestivum, Gluten protein, High-performance liquid chromatography, Qualitative and quantitative analyses

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