Elimination Mechanism Of Alanine And Serine For Acrolein And The Cytotoxicity Of Their Interaction Products

Acrolein is a highly reactive ?,?-unsaturated aldehyde,which is widely present in the environment,food processing and organisms.Preliminary research in the laboratory found that amino acids can react with compounds containing carbonyl groups to reduce the toxicity of dicarbonyl compounds.At the same time,many studies have found that theoretically the acrolein content produced during the frying process of snack food potato chips is 2 to 3 orders of magnitude higher than the acrolein content detected in the actual fried potato chips,and the amino acid content is also greatly reduced during this process.We speculate that the two can react to form other substances during the processing,which reduces the content of acrolein.In this study,the main amino acids in potato were reacted with acrolein to prepare and characterize the main products,and the quantitative detection method was established to determine the content of adducts in potato chips.Finally,taking alanine and serine as examples,the effects of the interaction products of amino acids and acrolein on the cytotoxicity of the three cell lines were studied,and the effect of the formation of the interaction products on cell apoptosis and its mechanism were discussed.The main contents are as follows:1.The elimination capacity of alanine and serine for acroleinUnder different reaction conditions,the elimination efficiency of alanine and serine on acrolein and the production of products in the reaction were studied,and it was found that both amino acids can efficiently eliminate acrolein.At different temperatures,both alanine and serine can effectively eliminate acrolein,and temperature,p H,and reaction ratio will all affect the elimination efficiency of amino acids on acrolein.Comparing the formation of products after the reaction of amino acids and acrolein at different temperatures(37 ?,50 ?,160 ?),it is found that the reaction products formed at different temperatures are different,and the lower temperature(37 ?,50 ?)mainly produces one kind of reaction product,the maximum absorption wavelength of the interaction product is 220 nm,and another brand-new product will be generated at the thermal processing temperature(160 ?),the maximum absorption wavelength is 195 nm.2.Identification of the products formed between amino acids and acrolein,and their detection in foodsBased on the elimination effect of amino acids on acrolein under different conditions,a reaction ratio of amino acid and acrolein with 2:1 was choosed to prepare adducts.And 9 kinds of amino acids with high content in potato were selected to prepare their interaction products with acrolein,and the reaction mixture was separated and purified by the combination of ODS and LH-20 to obtain the target product with a purity of more than 93%.After structural identification of the product by HPLC-MS/MS,HRMS,and NMR,it was found that the interaction products of amino acids and acrolein mostly showed cyclic or polyunsaturated structures,and the main reaction mechanism of amino acids and acrolein was summarized: michael addition Formation reaction and carbonyl ammonia reaction are the main ways for amino acids to eliminate acrolein.The nitrogen-containing six-membered ring is a stable structure that tends to form at low and high temperatures.A quantitative detection method for the amino acid-acrolein interaction products in foods was established by using high-performance liquid-phase mass spectrometry(HPLC-MS/MS),and the product of alanine,serine,asparagine with acrolein in two different potato chips were determined,the content of them was ranging from 1 to100 ?g/kg,which has a relatively large relationship with the type of amino acid.3.Cytotoxicity evaluation of amino acid-acrolein reaction productsThe cytotoxicity of the reaction mixture,purified adducts and acrolein on GES-1 cells,Caco-2 cells and HUVEC cells were studied respectively.The results showed that acrolein has greater toxic and side effects on the three kinds of cells.After 24 hours of culture,the IC50 of the cells were 71.81 ?M(GES-1 cells),53.66 ?M(Caco-2 cells),and 16.20 ?M(HUVEC cells).Even if the separated and purified interaction products are processed at the maximum concentration,the cell survival rate remains at nearly 80% and above.By detecting the level of oxidative stress,DNA damage,and apoptosis in cells,it was found that acrolein can induce oxidative stress by upregulating the levels of ROS in three kinds of cells,and at the same time induce cell nuclear condensation,affect DNA synthesis and cause DNA Fragmentation induces apoptosis and increases cell mortality,and the formation of the interaction product of amino acids and acrolein can reduce the toxicity of acrolein by reducing the level of intracellular oxidative stress,reducing DNA damage and cell apoptosis.