碘量法快速筛查多种β-内酰胺酶抑制剂  

Rapid Screening of β-Lactamase Inhibitors by Iodometric Method

作　　者：吴晓丹 刘凤银 王小鹏 陈凤燕 陈少华 袁学文 穆洪涛 WU Xiaodan;LIU Fengyin;WANG Xiaopeng;CHEN Fengyan;CHEN Shaohua;YUAN Xuewen;MU Hongtao(College of Biology and Food Engineering,Guangdong University of Education,Guangzhou 510303,China;Guangdong Provincial Bioengineering Institute,Guangzhou Sugarcane Industry Research Institute,Guangzhou 510303,China)

机构地区：[1]广东第二师范学院生物与食品工程学院,广东广州510303 [2]广东省生物工程研究所(广州甘蔗糖业研究所),广东广州510303

出　　处：《食品科技》2020年第10期307-316,共10页Food Science and Technology

基　　金：国家自然科学基金项目(31601540);广东省青年创新人才类基金项目(2017KQNCX142);广东第二师范学院大学生创新创业计划训练项目(201914278132)。

摘　　要：旨在建立一种碘量法,用于对多种β-内酰胺酶抑制剂的快速筛查。以舒巴坦为基准药物,采用全波长扫描确定最佳测定波长,棋盘滴定法及单因素实验确定最佳工作条件,绘制舒巴坦标准曲线,通过测定药物的交叉反应率,考察了该方法对4种常见β-内酰胺酶抑制剂的检测灵敏度。结果显示,最佳工作条件为测定波长585 nm、青霉素酶浓度1.0×104 U/mL、青霉素G钾浓度0.2 g/L、碘液浓度0.3 g/L、淀粉浓度1%、反应pH6.0、反应温度37℃、孵育时间20 min、催化时间30 min;对舒巴坦的检出限(LOD,IC10)为410.13μg/kg,半抑制浓度(IC50)为2522.08μg/kg,线性范围(IC20~IC80)为801.79~7933.30μg/kg;对克拉维酸、他唑巴坦、阿维巴坦的交叉反应率分别为454.17%、574.30%、2319.79%,检出限分别为224.64、67.56、42.86μg/kg。该研究提供了碘量法检测β-内酰胺酶抑制剂体系的优化方法,可实现在60 min内完成对多种β-内酰胺酶抑制剂的快速筛查。An iodometric method was developed for rapid screening ofβ-lactamase inhibitors.With sulbactam as the benchmark drug,the best measuring wavelength was determined by full wavelength scanning,and the optimum working conditions were determined by chessboard titration method and single factor experiment,and then the standard curve of sulbactam was established.The sensitivity of the method to four commonβ-lactamase inhibitors was investigated by measuring the cross-reactivity.The optimum working conditions were as follows:measuring wavelength 585 nm,the concentration of penicillinase 1.0×104 U/mL,penicillin G potassium 0.2 g/L,iodine solution 0.3 g/L,starch 1%,reaction pH 6.0,temperature 37℃,incubation time 20 min,and catalytic time 30 min.The proposed method exhibited the limit of detection(LOD,IC10)at 410.13μg/kg,median inhibitory concentration(IC50)at 2522.08μg/kg,and linear range(IC20~IC80)of 801.79~7933.30μg/kg for sulbactam.The cross-reactivity of clavulanic acid,tazobactam and avibactam were 454.17%,574.30%and 2319.79%,respectively,and the limits of detection were 224.64,67.56,42.86μg/kg,respectively.In this research,a systematic optimization method for the detection ofβ-lactamase inhibitors by iodometric method was provided,and a variety ofβ-lactamase inhibitors can be rapid screened by this method within 60 minutes.

关 键 词：碘量法 Β-内酰胺酶抑制剂 青霉素酶 青霉素G 

分 类 号：TS207.5+3[轻工技术与工程—食品科学]