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比较高效液相色谱法(HPLC)和毛细管区带电泳法(FZCE)在小麦属聚类分析及硬粒小麦掺假鉴定中的应用。

Compared use of HPLC and FZCE for cluster analysis of Triticum spp and for the identification of T. durum adulteration.

作者信息

Bonetti Alessandra, Marotti Ilaria, Catizone Pietro, Dinelli Giovanni, Maietti Annalisa, Tedeschi Paola, Brandolini Vincenzo

机构信息

Department of Agroenvironmental Science and Technology (DiSTA), University of Bologna, V.le Fanin 44, 40127, Bologna, Italy.

出版信息

J Agric Food Chem. 2004 Jun 30;52(13):4080-9. doi: 10.1021/jf034881f.

Abstract

Wheat quality criteria continually evolve in response to market pressure and consumer preference. Characterization of cereal cultivars for quality and agronomic properties, have widely shown the importance of the protein content to ensure good quality products. The aim of this work is a comparison of reversed-phase high performance liquid chromatography (RP-HPLC) and free zone capillary electrophoresis (FZCE) in the identification of Italian wheat cultivars and detection of durum wheat flour adulteration. Mainly alcohol soluble (gliadins) and water soluble (albumins) proteins were extracted from 14 common wheat cultivars and from 9 durum wheat cultivars. In RP-HPLC chromatograms, wheat albumins and gliadins eluted between 3 and 9 min and between 10 and 42 min, respectively. Even if the chosen chromatographic conditions (reversed phase) did not permit a complete resolution of hydrophilic proteins such as albumins, a good reproducibility was observed for both albumins and gliadins. In FZCE electropherograms, wheat albumins and gliadins migrated between 8 and 14 min and 16-25 min, respectively. A good reproducibility was found for wheat albumins, while the relatively poor reproducibility of gliadin fractions was a consequence of the selected separation conditions aimed to separate in the same run either hydrophilic (albumins) and alcohol-soluble (gliadins) proteins. The principal component analysis (PCA) of HPLC and FZCE data evidenced that both techniques allowed the univocal identification of the great proportion of investigated wheat cultivars. Three peaks were exclusively detected in RP-HPLC chromatograms of common wheat cultivars, while three unique peaks were found in FZCE electropherograms of common wheat cultivars. These peaks were investigated as a basis for detecting and estimating the adulteration of durum wheat flour with flour from common wheat. The direct relationship between the area of the peaks and adulteration level enabled standard curves to be constructed. The standard curves showed that adulteration may be quantified by either RP-HPLC or FZCE.

摘要

小麦品质标准会随着市场压力和消费者偏好不断演变。对谷物品种的品质和农艺特性进行表征,已充分表明蛋白质含量对于确保优质产品的重要性。本研究的目的是比较反相高效液相色谱法(RP-HPLC)和自由区毛细管电泳法(FZCE)在鉴定意大利小麦品种以及检测硬粒小麦粉掺假方面的效果。主要从14个普通小麦品种和9个硬粒小麦品种中提取醇溶蛋白(麦醇溶蛋白)和水溶蛋白(清蛋白)。在RP-HPLC色谱图中,小麦清蛋白和麦醇溶蛋白分别在3至9分钟和10至42分钟之间洗脱。即使所选的色谱条件(反相)无法完全分离清蛋白等亲水性蛋白质,但清蛋白和麦醇溶蛋白均具有良好的重现性。在FZCE电泳图中,小麦清蛋白和麦醇溶蛋白分别在8至14分钟和16至25分钟之间迁移。小麦清蛋白具有良好的重现性,而麦醇溶蛋白组分重现性相对较差,这是由于所选分离条件旨在在同一运行中分离亲水性(清蛋白)和醇溶性(麦醇溶蛋白)蛋白质。对HPLC和FZCE数据进行主成分分析(PCA)表明,两种技术都能明确鉴定出大部分被研究的小麦品种。在普通小麦品种的RP-HPLC色谱图中仅检测到三个峰,而在普通小麦品种的FZCE电泳图中发现了三个独特的峰。这些峰被作为检测和估算硬粒小麦粉中掺有普通小麦粉的依据进行研究。峰面积与掺假水平之间的直接关系使得能够构建标准曲线。标准曲线表明,掺假情况可以通过RP-HPLC或FZCE进行定量。

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