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提高动物副产品的真实性:新型成分DNA提取方法的协调统一。

Enhancing the authenticity of animal by-products: harmonization of DNA extraction methods from novel ingredients.

作者信息

Filipa-Silva Andreia, Castro Raquel, Rebelo Mariana, Mota Maria J, Almeida André, Valente Luísa M P, Gomes Sónia

机构信息

CIIMAR/CIMAR-LA, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Matosinhos, Portugal.

ICBAS, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal.

出版信息

Front Chem. 2024 Feb 20;12:1350433. doi: 10.3389/fchem.2024.1350433. eCollection 2024.

Abstract

The increasing global pressure to explore alternative protein sources derived from animal by-products has opened-up opportunities, but it has also created the need to assess their compliance with labelling statements, to ensure consumer's trust in the composition of both feed and food products. Assessing the authenticity of highly processed animal by-products, particularly within the rapidly expanding Halal food market, presents a significant challenge due to the lack of robust and standardized methodologies. However, the success of DNA based authenticity system is highly dependent on the extracted DNA quantity, quality, and purity ratios from heterogeneous matrices. In this work, nine DNA extraction methods were tested on selected processed animal by-products with high-value and interest for the feed industry: meals from poultry meat, blood and feather, and hydrolysates from swine meat and bone, fish, and black soldier fly. The proposed DNA extraction methods are developed to specifically target swine-specific mitochondrial region, as a case study. Both the conventional CTAB method and the commercial kits, specifically Invisorb Spin Tissue Mini and NucleoSpin™ Food, demonstrated superior extraction efficiency and quality ratios. Nevertheless, commercial kits enabled faster detection in comparison to the conventional methods. The absence of swine DNA was successfully validated and confirmed in all animal meals and hydrolysates that did not contain swine in their composition beforehand, demonstrating their compliance with the Halal market requirements.

摘要

探索源自动物副产品的替代蛋白质来源所带来的全球压力与日俱增,这既带来了机遇,也使得有必要评估这些产品是否符合标签声明,以确保消费者对饲料和食品产品成分的信任。由于缺乏可靠且标准化的方法,评估高度加工的动物副产品的真实性,尤其是在迅速扩张的清真食品市场中,是一项重大挑战。然而,基于DNA的真实性系统的成功高度依赖于从异质基质中提取的DNA数量、质量和纯度比例。在这项工作中,对九种DNA提取方法进行了测试,所选的加工动物副产品对饲料行业具有高价值且备受关注:禽肉、血液和羽毛制成的饲料,以及猪肉和骨头、鱼类和黑水虻的水解产物。作为案例研究,所提出的DNA提取方法专门针对猪特异性线粒体区域进行开发。传统的十六烷基三甲基溴化铵(CTAB)方法和商业试剂盒,特别是Invisorb Spin Tissue Mini和NucleoSpin™ Food,均展现出卓越的提取效率和质量比例。尽管如此,与传统方法相比,商业试剂盒能够实现更快的检测。在所有事先成分中不含猪的动物饲料和水解产物中,成功验证并确认了不存在猪DNA,这表明它们符合清真市场的要求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c71d/10912508/9ae7bf978820/fchem-12-1350433-g001.jpg

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