新橙皮苷二氢查尔酮化学合成工艺改进及其体外降脂活性研究  

Process optimization of neohesperidin dihydrochalcone synthesis and its in vitro hypolipidemic activity

作　　者：赖金娥 汤赫 曹庸[3] 曾灿彪 胡锋 陈永生[1] 晏日安[1] LAI Jin’e;TANG He;CAO Yong;ZENG Canbiao;HU Feng;CHEN Yongsheng;YAN Ri’an(Department of Food Science and Engineering,Jinan University,Guangzhou 510632,China;Zhangjiajie Mingyuan Biological Technology Co.Ltd.,Zhangjiajie 427000,China;College of Food Science,South China Agricultural University,Guangzhou 510642,China;Guangdong Chubang Food Co.Ltd.,Yangjiang 529800,China)

机构地区：[1]暨南大学食品科学与工程系,广东广州510632 [2]张家界铭源生物科技有限公司,湖南张家界427000 [3]华南农业大学食品学院,广东广州510642 [4]广东厨邦食品有限公司,广东阳江529800

出　　处：《食品与发酵工业》2024年第21期189-196,共8页Food and Fermentation Industries

基　　金：二氢查尔酮合成小试技术;阳西县科技计划项目(202101)。

摘　　要：新橙皮苷二氢查尔酮(neohesperidin dihydrochalcone,NHDC)是非营养型甜味剂,可用于无糖食品。该研究以化学方法合成NHDC并探究其体外的胰脂肪酶抑制效果。以廉价易得的柚皮苷为原料,经碱性条件开环得到分解产物根皮乙酰苯-4′-新橙皮苷(phloroacetophenone-4′-neohesperidoside,PN),通过Claisen-Schmidt缩合反应合成中间体新橙皮苷,再以廉价的雷尼镍替代昂贵的Pd/C催化氢化反应,合成目标产物NHDC。以高分辨质谱仪、核磁共振波谱仪鉴定产物结构。通过体外胰脂肪酶(pancreatic lipase,PL)抑制实验、荧光猝灭实验探究其降脂活性与动力学作用机制。结果表明,PN的合成中,NaOH与柚皮苷的最佳质量比为1.7,合成新橙皮苷的催化剂吡咯烷与乙酸的最佳摩尔比为2∶1,合成NHDC的最佳NaOH质量分数为7%,雷尼镍催化能力强于Pd/C,最佳添加量为20%。合成工艺改进后,NHDC的总收率为70.6%。NHDC能抑制PL的活性,半数抑制浓度(IC_(50))为0.71 mg/mL,抑制类型为可逆、反竞争性抑制,NHDC与PL结合后,猝灭PL的内源荧光,改变酶结构及微环境,从而降低PL活性,荧光猝灭类型为混合猝灭。因此,NHDC拥有作为胰脂肪酶抑制剂的应用潜力,具有减少脂质吸收的作用。Neohesperidin dihydrochalcone(NHDC)is a non-nutritive sweetener that can be used in sugar-free foods.This study involved the chemical synthesis of NHDC and an investigation of its pancreatic lipase inhibitory effect in vitro.The intermediate neohesperidin was synthesized using inexpensive and readily available naringin as a raw material.The decomposition product,phloroacetophenone-4′-neohesperidoside(PN),was obtained through alkaline condition ring-opening,and neohesperidin was synthesized via a Claisen-Schmidt condensation reaction.The catalytic hydrogenation reaction was carried out by replacing the expensive Pd/C with cheap Rainey nickel,resulting in the synthesis of the target product NHDC.The structure of the product was identified using high-resolution mass spectrometry and a nuclear magnetic resonance spectrometer.In vitro,pancreatic lipase(PL)inhibition assay and fluorescence quenching assay were used to explore its lipid-lowering activity and kinetic mechanism.The results indicated that the optimum mass ratio of sodium hydroxide to naringin for the synthesis of PN was 1.7,the optimum molar ratio of pyrrolidine to acetic acid for the catalyst for the synthesis of neohesperidin was 2∶1,the optimum sodium hydroxide concentration for the synthesis of NHDC was 7%,and the optimum amount of Raney nickel,which exhibited stronger catalytic ability than Pd/C,was added at a level of 20%.The total yield of NHDC was 70.6%after the improved synthesis process.NHDC was found to inhibit the activity of PL,with a half inhibitory concentration(IC_(50))of 0.71 mg/mL,and the inhibition type was reversible,anti-competitive inhibition.After NHDC combined with PL,it quenched the endogenous fluorescence of PL and changed the enzyme structure and microenvironment,thereby reducing PL activity.The fluorescence quenching type was identified as mixed quenching.Thus,NHDC has the potential to be applied as a pancreatic lipase inhibitor with the ability to reduce lipid uptake.

关 键 词：新橙皮苷二氢查尔酮 胰脂肪酶 化学合成 荧光猝灭 

分 类 号：TS201.2[轻工技术与工程—食品科学]