Effect Of 3-aminopropanamide On The Reduction Of Acrylamide

3-Aminopropanamide?3-APA?is an intermediate product which forms via the Maillard reaction between asparagine and reducing sugars.It is a direct precusor of acrylamide and can produce acrylamide through deamination at high temperature.In this research,we found that3-APA eliminated acryalmide via the formation of adduct.Based on this finding,the cytotoxicity of adduct 2 towards Caco-2 cell line and the effect of several food components on the elimination efficiency of 3-APA for acrylamide were investigated.The main conclusions were as follows:?1?3-Aminopropanamide reduced acrylamide through Michael addition reactionWhen heating 3-APA or 3-APA along with acrylamide at 160?for 20 min,two new peaks A1 and A2 on HPLC were detected in the reaction products.The molecular weight of A1and A2 were 159 and 230 detected by using TOF-MS and HPLC-MS.The result of TOF-MS showed that their molecular formula was C₆H₁₃N₃O₂ and C₉H₁₈N₄O₃,respectively.The retention time of A2 is the same to that of 3,3?,3?-nitrilotris,and its MS-MS fragments were consistent with that of 3,3?,3?-nitrilotris,it consist of one molecule of acrylamide and two molecules of3-APA.Thus,A2 was regarded as 3,3?,3?-nitrilotris.The level of adducts was significantly affected by temperature,heating time and the ratio of reactants.When the concentration ratio of acrylamide to 3-aminopropanamide was kept at3:5 and reacted at 160?for 20 min,47.29%of acrylamide was eliminated through Michael addition reaction.?2?The adduct showed lower cytotoxicity than acrylamide against Caco-2 cells.When Caco-2 cells were incubated with 16 mM acrylamide for 24 h and 48 h,the cell viabilities were 23.3%and 19.1%,respectively.However,the cell viabilities incubated with 64m M adduct 2 were still kept at 83.0%and 81.1%,respectively.Addition of acrylamide had a significant effect,but the adduct showed no significant on the cell morphology and the cell cycle of Caco-2 cells.Some cells became round,shrinking and the dead cells were fallen off after the addition of acrylamide.As the concentration of acrylamide increased,change became more dramatically.What's more,the ratio of G1 period slightly increased when the concentration of acrylamide was 0.5 mM,and then the ratio of G1 period reduced while the ratio of G2 period increased as the concentrations of acrylamide increased.When the concentrations of acrylamide were 0.5 and 1 mM,respectively,Caco-2 cells mainly stopped in the G1 period.When the concentrations of acrylamide were 2 and 4 mM,respectively,Caco-2cells mainly stopped in the G2 period.Otherwise,the concentrations of acrylamide measured in the medium which treated with adduct 2 were less than 3%.So,3-aminopropanamide could reduce the cytotoxicity of acrylamide through Michael addition reaction.?3?Effect of glucose,caffeic acid and quinic acid on the elimination of acrylamide by 3-APAThe reactivity of amine group of 3-APA is similar to that of amino acids,which can react with reducing sugars and influence the conversion of acrylamide from 3-APA.The addition of glucose significantly increased acrylamide content produced from 3-APA,and only adduct 1was detected in 3-APA/Glucose reaction system.The result indicated that the addition of glucose increased the formation of acrylamide and inhibited acrylamide elimination.Caffeic acid and quinic acid were added into 3-APA/Glucose reaction system in order to investigate their effect on the conversion of acrylamide form 3-APA.It was found that their concentrations,heating temperature and time can influence the conversion of acrylamide from 3-APA in3-APA/glucose reaction system.Two possible mechanisms on the influence of acrylamide formation by caffeic acid and quinic acid were speculated.Firstly,3-APA was consumed by the degradation products of caffeic acid and quinic acid.Secondly,polymerization of acrylamide was promoted by the free radicals which formed from degradation of caffeic acid and quinic acid.