DNA Extraction And Traceability Analysis For Edible Vegetable Oil Based On Non-silica-based Nanoparticles Technology

Edible vegetable oil is an important part of people's daily dietary nutrition.It can provide calories and some essential fatty acids for the human body.But driven by economic interests,edible vegetable oil adulteration occurs frequently,which not only disturbs the market order,infringes on consumer rights and interests,but also endangers people's health.Authenticity identification technology of edible vegetable oils is an advanced science technology to crack down on oil adulteration,safeguard consumers' rights and interests and market order,and strengthen government supervision.At present,the most popular detection technologies in the market are physicochemical detection methods,such as spectrophotometry,mass spectrometry,chromatography and their mutual use.In recent years,reports about the use of biological methods to detect adulteration of edible vegetable oil have been issued.Because biological methods are simple,rapid,sensitive and so on,they have gradually become a research hotspot of edible vegetable oil detection technology.However,in the detection of edible vegetable oils,many biological detection methods based on molecular techniques are not very mature.The most critical reason is that it is difficult to extract enough high-quality DNA in deep-processed edible vegetable oil.In this study,molecular biological methods were used to extract DNA from vegetable oil with new nanomaterials,according to the difference between species and the conservation within species of DNA,species traceability identification and adulteration identification were carried out by real-time fluorescent quantitative PCR assay of specific DNA fragments.The main findings are as follows:(1)The novel nanoparticle material used for the extraction of refined edible vegetable oil DNA,is composed of dipolar polystyrene-polymethyl methacrylate complex nanoparticles(DPPS-PMNP).This nanoparticle has unique hydrophilicity,lipophilicity and ultra-high adsorption and desorption of DNA.(2)The extraction efficiency of DPPS-PMNP particles in edible vegetable oil has a certain relationship with the ratio of the amount of DPPS-PMNP nanospheres and the amount of DNA in the system.The study found that when the ratio of the two was 5:1,the recovery efficiency was the highest,reaching 76.56%.(3)The DNA quality of refined edible vegetable oil extracted by DPPS-PMNP Kit is superior to the other two commercially available kits,and can be used as a real-time PCR template for subsequent traceability analysis experiments.(4)The specific primers and probes for the detection of peanut,soybean,rapeseed and cotton were designed with plant species-specific chloroplast ribulose-1,5-diphosphate gene rbcL as the target gene,the amplification efficiencies reached 96.9%,82.3%,95.7%and 91.7%,respectively.(5)DPPS-PMNP Kit was used to extract sufficient amount of DNA from pure edible vegetable oil and various mixed edible vegetable oil.Through real-time fluorescence quantitative PCR traceability analysis,not only the plant species composition in the mixed oil could be detected,but also the mixture proportion of each species of vegetable oil could be detected.The above results showed that the DPPS-PMNP nanoparticle vegetable oil DNA extraction and quantitative PCR detection kit could extract sufficient and high purity DNA from pure edible vegetable oil and mixed vegetable oil,and perform species traceability identification and quantitative analysis of mixed oil for edible vegetable oil.