微滴式数字PCR定量检测杏仁露中杏仁、花生源性成分

Quantitative Detection of Apricot Kernel-derived and Peanut-derived Components in Apricot Kernel Protein Drink by Droplet Digital PCR

为定量分析市售杏仁露中杏仁源性成分和掺假花生源性成分的含量,本研究基于微滴式数字聚合酶链式反应(droplet digital polymerase chain reaction,ddPCR)技术建立准确的检测方法。从杏仁、花生的植物组织提取核酸后,建立植物源性成分质量(M)与DNA拷贝数(C)之间的线性关系。ddPCR研究结果显示,在一定范围内杏仁、花生的质量和DNA含量、DNA含量和DNA拷贝数均呈显著的线性关系,以DNA含量为中间值换算得出公式:M杏仁=0.13C+1.24,M花生=0.081C-0.63。通过已知混合比例的两物种掺假模型验证该公式的准确性和适用性,并对市售的12个杏仁露样品进行检测。研究表明,该方法准确可靠,能达到精准定量。本研究建立的ddPCR方法可有效甄别杏仁露中的花生源性成分为无意沾染或有意掺杂,在杏仁露中杏仁和花生含量的定量检测方面具有较大应用潜力,为植物蛋白饮料的市场监管提供技术支持。

An accurate method to quantitatively analyze the contents of apricot kernel-derived and peanut-derived components in apricot kernel protein drink products was established based on droplet digital polymerase chain reaction(ddPCR) in this study. We established a linear relationship between plant material mass(M) and DNA copy number(C) after the extraction of nucleic acids from both apricot kernels and peanuts. According to the results of ddPCR, we found that the relationships between the mass of each plant material and DNA concentration and between DNA concentration and DNA copy number were both significantly linear within a certain range. The DNA concentration was utilized as an intermediate value to establish the following formulae: Mapricot kernel = 0.13 C + 1.24, Mpeanut = 0.081 C-0.63. The accuracy and applicability of this method were tested and verified using known mixtures of apricot kernel and peanut and 12 commercial apricot protein drink products. This method enabled accurate and reliable quantitative analysis, and effective identification of whether the adulteration in apricot kernel protein drink products is inadvertent or intentional. Hence ddPCR has strong potential in quantifying apricot kernel and peanut in apricot kernel protein drink products and provides technical support for market supervision of plant protein drink products.