Hernández Alejandro, Martín Alberto, Aranda Emilio, Bartolomé Teresa, Córdoba María de Guía
Nutrición y Bromatología, Escuela de Ingenierías Agrarias, Universidad de Extremadura, Ctra. de Caceres s/n, 06071 Badajoz, Spain.
J Agric Food Chem. 2006 Jun 14;54(12):4141-7. doi: 10.1021/jf060349r.
The purpose of this work was to develop a procedure based on protein analysis by free zone capillary electrophoresis (FZCE) that can be used in the determination of smoked paprika "Pimentón de La Vera" adulteration with paprika elaborated from varieties of pepper foreign to the "La Vera" region, in central western Spain. Two autochthonous varieties of pepper, Jaranda and Bola, and the variety Papri Queen, foreign to the "La Vera" region, were used in the study. Several aqueous solutions for solubilization of the methanol-soluble proteins were tested, and the FZCE conditions of capillary dimensions, FZCE buffer concentrations, and detection wavelengths were optimized. On the basis of the results, 30% (v/v) acetonitrile was adopted as the suspending solution for routine analysis, and the optimal FZCE parameters were 75 microm inner diameter and 57 cm total length capillaries, 8.75 mM phosphate/20.6 mM tetraborate as run buffer, and 256 nm as detection wavelength. This method was found to give excellent repeatability of the corrected migration time (CMT) with coefficients of variation (RSD %; n = 5) of <1% for most of the proteinaceous compounds analyzed and showed greater effectiveness in discriminating paprika varieties than the SDS-PAGE technique. Four peaks found in the FZCE electropherograms were investigated as a basis for detecting and estimating the adulteration of smoked paprika with paprika elaborated from the Papri Queen variety. The adulteration detection limits varied from 5 to 40% of the Papri Queen variety within a satisfactory working range of mixture (5-80%) sufficiently large to cover the adulteration levels of interest. The use of peak 6 as a marker for determining adulteration gave the best results, with an adulteration detection limit of 5-10% (w/w).
本研究旨在开发一种基于自由区毛细管电泳(FZCE)蛋白质分析的方法,用于检测西班牙中西部“拉韦拉”地区烟熏辣椒粉“Pimentón de La Vera”是否掺假,掺假物为用该地区以外辣椒品种制成的辣椒粉。研究使用了两种当地辣椒品种Jaranda和Bola,以及一种非“拉韦拉”地区的Papri Queen品种。测试了几种用于溶解甲醇溶性蛋白质的水溶液,并优化了毛细管尺寸、FZCE缓冲液浓度和检测波长等FZCE条件。根据结果,采用30%(v/v)乙腈作为常规分析的悬浮液,最佳FZCE参数为内径75微米、总长57厘米的毛细管,8.75 mM磷酸盐/20.6 mM硼酸盐作为运行缓冲液,256 nm作为检测波长。该方法对校正迁移时间(CMT)具有出色的重复性,大多数分析的蛋白质化合物的变异系数(RSD%;n = 5)<1%,并且在区分辣椒粉品种方面比SDS-PAGE技术更有效。对FZCE电泳图中发现的四个峰进行了研究,作为检测和估计烟熏辣椒粉中掺有Papri Queen品种辣椒粉的依据。在令人满意的混合工作范围(5-80%)内,掺假检测限为Papri Queen品种的5-40%,该范围足够大,能够涵盖感兴趣的掺假水平。使用峰6作为确定掺假的标记物效果最佳,掺假检测限为5-10%(w/w)。