摘要
目的建立截短适配体-荧光法测定水中双酚A。方法选择包含38个碱基的双酚A截短适配体为双酚A识别元件,在截短适配体的5’端修饰荧光基团6-FAM,在互补序列c DNA的3’端修饰淬灭基团DABCYL,配制双酚A及干扰物标准溶液,优化互补序列c DNA的碱基个数、截短适配体与互补序列cDNA的浓度比、孵育温度、孵育时间和缓冲液pH值,建立截短适配体-荧光法检测体系,进行特异性与回收率实验。结果当互补序列cDNA包含9个碱基,截短适配体与互补序列cDNA的浓度比为1∶1.5,缓冲溶液p H值为7.5,55℃孵育60 min时,10~75 pmol/L浓度范围内,双酚A的线性关系较好,线性回归方程为y=2 230.7x+110 825,相关系数为0.926,检出限为3.3 pmol/L。四溴双酚A、雌二醇、雌三醇和双酚S等4种干扰物与双酚A的荧光强度差值区别较明显,对检测结果未造成明显干扰。当加标浓度为20.0、40.0和60.0 pmol/L时,回收率分别为97.8%、98.8%和102.3%,相对标准偏差分别为4.4%、2.1%和2.6%。结论建立的截短适配体-荧光法操作简便,灵敏度高,特异性强,可用于检测水中双酚A。
ObjectiveTo establish a fluorescence method based on turncated aptamer for the determination of bisphenol A in water.MethodsThe bisphenol A truncated aptamer containing 38 bases was selected as a recognition module,and was modified with the fluorophore 6-FAM at the 5’end. The 3’end of the complementary sequence cDNA was modified with the quencher DABCYL. The standard solutions of bisphenol A and interfering compounds were configured.The detection system was established after optimizing the number of bases in c DNA, the concentration ratio of truncated aptamer to cDNA, the incubation temperature and time, and the p H of the buffer. The specificity and recovery experiments were carried out.ResultsWhen the complementary sequence c DNA included 9 bases, the concentration ratio of the truncated aptamer to cDNA was 1∶1.5, the p H value of the buffer solution was 7.5, the cDNA was incubated at 55℃for 60 minutes, in the concentration range of 10-75 pmol/L, the linear regression equation was y=2 230.7 x+110 825,the correlation coefficient was 0.926. The limits of detection was 3.3 pmol/L. The difference values of fluorescence intensity between tetrabromobisphenol A, estradiol, estriol, bisphenol S and bisphenol A were obviously different, so there was no significant interference to the test result. The recovery rates were 97.8%, 98.8% and 102.3% with the spiked concentrations of 20.0, 40.0 and 60.0 pmol/L. The relative standard deviations were 4.4%, 2.1% and 2.6%(n=5), respectively.Conclusion high sensitivity and specificity, which can be used for the determination of bisphenol A in water.
作者
薛晨晨
朱光平
白洁
吴南翔
范宏亮
XUE Chenchen;ZHU Guangping;BAI Jie;WU Nanxiang;FAN Hongliang(School of Public Health,Hangzhou Medical College,Hangzhou,Zhejiang 310013,China)
出处
《预防医学》
2021年第11期1086-1090,共5页
CHINA PREVENTIVE MEDICINE JOURNAL
基金
国家自然基金青年基金(21904118)
浙江省自然科学基金(LY17B050008)。
关键词
截短适配体
荧光法
检测
双酚A
truncated aptamer
fluorescence analysis
detection
bisphenol A
