基于“碘-罗丹明B”缔合物荧光探针测定β-内酰胺酶活性  

Determination ofβ-lactamase activity using iodine-rhodamine B associated content as a fluorescent probe

作　　者：戴兴德 雷林强 安军红 张小林 Dai Xing-de;Lei Lin-qiang;An Jun-hong;Zhang Xiao-lin(Gansu Medical College,Pingliang,Gansu 744000)

机构地区：[1]甘肃医学院,平凉744000

出　　处：《中国抗生素杂志》2020年第10期1040-1043,共4页Chinese Journal of Antibiotics

基　　金：甘肃省高等学校创新能力提升项目(No.2019A-159)。

摘　　要：碘(I3-)对罗丹明B(RB)具有荧光猝灭作用,可使RB的荧光信号强度减弱甚至消失,而β-内酰胺酶作用下的青霉素水解产物青霉噻唑酸可将I3-还原为I-,使体系荧光信号再现,据此建立了以碘-罗丹明B缔合物为荧光探针测定药剂中青霉素含量的新方法。在激发波长360nm,发射波长580nm条件下进行了β-内酰胺酶活性的荧光测定。结果表明,在0~2.4U/mL范围内,β-内酰胺酶活性与其荧光强度变化值呈良好的线性关系,检出限为0.002U/mL。方法的加标回收率为96.67%~103.3%,相对标准偏差(n=6)2.0%~4.5%。该方法简便快速、准确可靠、灵敏度高,成功用于自制酶液酶活性测定。When the fluorescence signal intensity of rhodamine B(RB)weakens or even disappears in the iodine(I3-)solution,penicillin thiazolic acid as a product of penicillin hydrolysis catalyzed by beta-lactamase can reduce I3-to I-,which makes the system fluorescent signal reappear.Based on this reaction,a method for determining theβ-lactamase activity by using the iodine-rhodamine B associate as a fluorescent probe was established.In this paper,fluorescence determination ofβ-lactamase activity was carried out at an excitation wavelength of 360nm and an emission wavelength of 580nm.The results showed that there was a good linear relationship betweenβ-lactamase activity and fluorescence intensity change in the range of 0~2.4U/mL.The limits of detection(LOD)was 0.002U/mL and the relative standard deviations(n=6)ranged at 2.0%~4.5%.The recovery rate was between 96.67%and 103.3%.Thus,the anti-fluorescence quenching method was simple,rapid,accurate,and reliable,and could be used for the determination of enzyme activity in self-prepared enzyme solution.

关 键 词：罗丹明B 碘 Β-内酰胺酶 荧光法 

分 类 号：R9[医药卫生—药学]