硫醇类生物标记物检测用香豆素衍生物的合成与应用

Synthesis and application of chromogenic reagentsfor detection of mercaptan biomarkers

以4-(二乙氨基)水杨醛为原料,无水氟化钾为催化剂,通过克脑文格尔反应制备化合物1,并对化合物1的合成条件进行优化,再通过尔斯迈尔-哈克-阿诺德反应和亲核加成-消除反应分别合成化合物2和化合物3[7-(二乙氨基)香豆素衍生物],以化合物3为显色剂,采用微流控纸芯片技术,实现对硫醇类生物标记物同型半胱氨酸(Hcy)、半胱氨酸(Cys)、谷胱甘肽(GSH)含量的特异性分析。结果表明,无水氟化钾可替代哌啶作为催化剂用于合成化合物1,m(无水氟化钾)∶m(反应物)=12%,n[4-(二乙氨基)水杨醛]∶n(乙酰乙酸乙酯)=1∶2,产率可达40.35%;化合物3分别与Cys、Hcy和GSH发生不同显色反应,可以实现对3种生物硫醇的特异性检测。

Using 4-diethylaminosalicylaldehyde as raw material and potassium fluoride as a catalyst,compound 1 was synthesized via a Knoevenagel reaction.The synthesis conditions of compound 1 were optimized.Then,compound 2 and 3(7-diethylaminocoumarin derivative)were synthesized through Ursmail-huck-arnold reaction and nucleophilic addition-elimination reaction,respectively.By employing compound 3 as a chromogenic agent,specific assays of homocysteine(Hcy),cysteine(Cys),and glutathione(GSH)were achieved using microfluidic paper-based analytical devices.The results show that anhydrous potassium fluoride can replace pyridine as a catalyst for the synthesis of compound 1.When the mass ratio of anhydrous potassium fluoride to the reactants is 12%,and the mole ratio of 4-diethylaminosalicylaldehyde to ethyl acetate is 1∶2,the yield of compuned 3 is 40.35%.Compound 3 reacts differently with Cys,Hcy,and GSH,enabling specific detection of three biologic thiols.