基于磁性分子印迹材料的荧光分光光度法测定血液中同型半胱氨酸的含量  

Determination of Homocysteine in Blood by Fluorescence Spectrophotometry Based on Magnetic Molecularly Imprinted Material

作　　者：信建豪[1] 侯巧芝[1] 王聪颖[1] 张利娟[1] 张蒙 XIN Jianhao;HOU Qiaozhi;WANG Congying;ZHANG Lijuan;ZHANG Meng(Medical College,Huanghe University of Science and Technology,Zhengzhou 450063,China)

机构地区：[1]黄河科技学院医学院,郑州450063

出　　处：《理化检验（化学分册）》2022年第10期1117-1121,共5页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：河南省科技攻关项目(172102310420);河南省重点学科工程项目(教高[2018]119号)。

摘　　要：提出一种基于磁性分子印迹材料的荧光分光光度法测定血液中同型半胱氨酸(Hcy)含量的方法。采用化学共沉淀法制备Fe_(3)O_(4)纳米磁芯,用硅酸四乙酯修饰后,以Hcy为模板分子,乙二醇二甲基丙烯酸酯为交联剂,偶氮二异丁腈为引发剂,在乙醇溶液中制备了Hcy纳米磁性分子印迹材料;用该材料吸附血液中的Hcy,释放Hcy后用衍生试剂N-(1-芘基)马来酰亚胺进行衍生,用荧光分光光度计测定体系的荧光强度。结果表明:Hcy纳米磁性分子印迹材料粒径约为40 nm;该材料对Hcy的吸附时间为10 min,吸附pH为3.0,最大表观吸附量为32.4 mg·g^(-1);对Hcy的释放时间为40 min,释放pH为8.0;加入N-(1-芘基)马来酰亚胺反应10 min后,以365 nm为激发波长,体系在387 nm处产生稳定荧光;并且该材料对Hcy有良好的选择性吸附,最多可重复使用4次;Hcy的质量浓度在0.02~0.10 mg·L^(-1)内与其对应的荧光强度呈线性关系,检出限为0.008 mg·L^(-1);对Hcy标准溶液进行精密度(n=5)和稳定性(n=6)试验,测定值的相对标准偏差均不大于4.0%;方法用于分析5只成年家兔的血液样品,结果均未检出Hcy;对上述空白样品进行加标回收试验,Hcy的回收率为91.2%~109%。A method for the determination of homocysteine(Hcy)in blood by fluorescence spectrophotometry based on magnetic molecularly imprinted material was proposed.Fe_(3)O_(4) nano magnetic core was prepared by chemical coprecipitation method,and modified with tetraethyl orthosilicatete.Then Hcy nano magnetic molecularly imprinted material was prepared in ethanol solution using Hcy as template molecule,ethylene glycol dimethacrylate as crosslinking agent and azodiisobutyronitrile as initiator.Hcy in blood was adsorbed by the material,the derivative reagent N-(1-pyrene)maleimide was used for derivation after releasing Hcy,and the fluorescence intensity of the system was determined by fluorescence spectrophotometer.It was shown that the particle size of Hcy nano magnetic molecularly imprinted material was about 40 nm.The adsorption time of Hcy was 10 min,with the adsorption pH of 3.0,the maximum apparent adsorption capacity of 32.4 mg·g^(-1),and the release time of Hcy was 40 min,with the release pH of 8.0.N-(1-pyrene)maleimide was added and reacted for 10 min.The stable fluorescence was generated at 387 nm with 365 nm as excitation wavelength.The material had good selective adsorption for Hcy and could be reused 4 times at most.Linear relationship between the fluorescence intensity and the mass concentration of Hcy was in the range of 0.02-0.10 mg·L^(-1),with detection limit of 0.008 mg·L^(-1).Tests for precision(n=5)and stability(n=6)were made on Hcy standard solution,and RSDs of the determined values were not more than 4.0%.This method was applied to analysis of the blood samples from 5 adult rabbits,and the results showed that Hcy was not detected.Test for recovery was made on the above blank samples,giving results in the range of 91.2%-109%.

关 键 词：同型半胱氨酸 磁性分子印迹材料 吸附 荧光强度 

分 类 号：O657.3[理学—分析化学]