Spectrophotometric Determination of Residual Amount of Tobramycin in Milk Based on Gold Nanoparticles Modified by Aptamer
摘 要
在没有妥布霉素存在时,适体包覆在金纳米粒子(AuNPs)表面可以防止盐诱导的聚集;在妥布霉素存在时,由于适体与妥布霉素的亲和力较高,适体与妥布霉素结合并从AuNPs表面脱离,导致AuNPs在适当含量的盐中发生聚集,反应体系颜色由红色变为蓝紫色。采用基于适体修饰的金纳米粒子分光光度法测定牛奶中妥布霉素的残留量。优化的试验条件如下:适体Hp孵育时间和妥布霉素孵育时间均为15 min;反应体系的pH为7;适体Hp的浓度为1.5 μmol·L-1;氯化钠的浓度为1.5 mol·L-1。妥布霉素的线性范围为40.0~175 nmol·L-1,检出限(3s/k)为13.3 nmol·L-1。以空白样品为基体进行加标回收试验,所得回收率为96.4%~108%,测定值的相对标准偏差(n=6)为1.8%~3.0%。
Abstract
In the absence of Tobramycin, the aptamer was coated on the surface of gold nanoparticles (AuNPs) to prevent salt-induced aggregation. In the presence of Tobramycin, the aptamer would separate from the surface of AuNPs and combine with Tobramycin because of the high affinity between the aptamer and Tobramycin, resulting in the aggregation of AuNPs at the appropriate content of salt, and the color of the reaction system changed from red to purple-blue. Spectrophotometry was applied to the determination of residual amount of Tobramycin in milk based on gold nanoparticles modified by aptamer. The optimized conditions found were as follows:incubation time of aptamer Hp and incubation time of Tobramycin were 15 min; pH of the reaction system was 7; concentration of aptamer Hp was 1.5 μmol·L-1; concentration of sodium chloride was 1.5 mol·L-1. Linearity range of Tobramycin was found between 40.0 nmol·L-1 and 175 nmol·L-1 with detection limit (3s/k) of 13.3 nmol·L-1. On the base of blank sample, test for recovery was made by standard addition method; values of recovery found were in the range of 96.4%-108%, with RSDs (n=6) of determined values in the range of 1.8%-3.0%.
中图分类号 O657.32 DOI 10.11973/lhjy-hx202102004
所属栏目 工作简报
基金项目 上海市国际科技合作基金项目(19230742900)
收稿日期 2019/12/3
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备注王燕,硕士研究生,主要从事纳米材料及其在食品安全检测方面的应用研究工作
引用该论文: WANG Yan,ZHOU Hualan,LIANG Yingfang,WANG Feng,SHI Qinyi. Spectrophotometric Determination of Residual Amount of Tobramycin in Milk Based on Gold Nanoparticles Modified by Aptamer[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2021, 57(2): 114~119
王燕,周化岚,梁营芳,王锋,施沁怡. 基于适体修饰的金纳米粒子分光光度法测定牛奶中妥布霉素的残留量[J]. 理化检验-化学分册, 2021, 57(2): 114~119
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参考文献
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【2】CHAUHAN R, SINGH J, SACHDEV T, et al. Recent advances in mycotoxins detection[J]. Biosensors and Bioelectronics, 2016,81:532-545.
【3】宣伟,王军,汪秀月,等.食品安全检测技术研究进展[J].肉类研究, 2011,25(9):47-51.
【4】ELLINGTON A D, SZOSTAK J W. In vitro selection of RNA molecules that bind specific ligands[J]. Nature, 1990,346:818-822.
【5】RUSCITO A, DEROSA M C. Small-molecule binding aptamers: Selection strategies, characterization,and applications[J]. Frontiers in Chemistry, DOI:10.3389/fchem.2016.00014.
【6】MA Q, WANG Y X, JIA J, et al. Colorimetric aptasensors for determination of tobramycin in milk and chicken eggs based on DNA and gold nanoparticles[J]. Food Chemistry, 2018,249:98-103.
【7】李爽,代昭,韩阳.金纳米粒子的合成方法及应用进展[J].精细石油化工, 2019,36(1):66-70.
【8】陈丹丹,辛嘉英,张兰轩,等.纳米金在食品安全检测中的应用[J].食品科学, 2014,35(7):247-251.
【9】NIE J J, YUAN L Y, JIN K, et al. Electrochemical detection of tobramycin based on enzymes-assisted dual signal amplification by using a novel truncated aptamer with high affinity[J]. Biosensors and Bioelectronics, 2018,122:254-262.
【10】FRENS G. Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions[J]. Nature Physical Science, 1973,241:20-22.
【11】NEW S Y, AUNG K M M, LIM G L, et al. Fast screening of ligand-protein interactions based on ligand-induced protein stabilization of gold nanoparticles[J]. Analytical Chemistry, 2014,86(5):2361-2370.
【12】SARDAR R, FUNSTON A M, MULVANEY P, et al. Gold nanoparticles: Past, present, and future[J]. Langmuir, 2009,25(24):13840-13851.
【13】SATO K, HOSOKAWA K, MAEDA M. Rapid aggregation of gold nanoparticles induced by non-cross-linking DNA hybridization[J]. Journal of the American Chemical Society, 2003,125(27):8102-8103.
【14】CHEN A L, JIANG X L, ZHANG W W, et al. High sensitive rapid visual detection of sulfadimethoxine by label-freeaptasensor[J]. Biosensors and Bioelectronics, 2013,42:419-425.
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