Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
J Agric Food Chem. 2011 Jan 26;59(2):513-20. doi: 10.1021/jf103702s. Epub 2010 Dec 23.
Usually spices are identified morphologically using simple methods like magnifying glasses or microscopic instruments. On the other hand, molecular biological methods like the polymerase chain reaction (PCR) enable an accurate and specific detection also in complex matrices. Generally, the origins of spices are plants with diverse genetic backgrounds and relationships. The processing methods used for the production of spices are complex and individual. Consequently, the development of a reliable DNA-based method for spice analysis is a challenging intention. However, once established, this method will be easily adapted to less difficult food matrices. In the current study, several alternative methods for the isolation of DNA from spices have been developed and evaluated in detail with regard to (i) its purity (photometric), (ii) yield (fluorimetric methods), and (iii) its amplifiability (PCR). Whole genome amplification methods were used to preamplify isolates to improve the ratio between amplifiable DNA and inhibiting substances. Specific primer sets were designed, and the PCR conditions were optimized to detect 18 spices selectively. Assays of self-made spice mixtures were performed to proof the applicability of the developed methods.
通常，香料的鉴别采用简单的形态学方法，如放大镜或显微镜。另一方面，聚合酶链反应（PCR）等分子生物学方法可以在复杂基质中进行准确和特异性检测。通常，香料的起源是具有不同遗传背景和关系的植物。生产香料所使用的加工方法复杂且独特。因此，开发一种可靠的基于 DNA 的香料分析方法是一个具有挑战性的目标。然而，一旦建立，这种方法将很容易适应于不太困难的食物基质。在本研究中，针对（i）纯度（分光光度法）、（ii）产量（荧光法）和（iii）扩增能力（PCR），详细开发和评估了几种从香料中分离 DNA 的替代方法。全基因组扩增方法用于预扩增分离物，以提高可扩增 DNA 与抑制物质之间的比例。设计了特异性引物对，并优化了 PCR 条件以选择性检测 18 种香料。进行了自制香料混合物的测定以证明所开发方法的适用性。
