1. 中国农业科学院农业质量标准与检测技术研究所
2. 四川农业大学食品学院
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牛书操, 吕珍珍, 刘金钏, 等. 基于适配体的纳米金比色法快速检测雌二醇研究[J]. 分析测试学报, 2014,33(7):835-839.
Rapid Detection of 17β-Estradiol Using Gold Nanoparticle Based Colorimetric Aptasensor[J]. 2014,33(7):835-839.
单链核酸适配体可以通过静电作用吸附到纳米金表面,保护纳米金免于高盐浓度引起的纳米金凝聚溶液变蓝现象;当加入靶标后,适配体与靶标结合,并从纳米金上解离,纳米金在高盐浓度下发生凝聚,溶液颜色由红变蓝。利用该原理,建立了一种基于适配体的雌二醇纳米金比色快速检测法。首先将雌二醇的适配体与纳米金室温孵育5 min,然后加入不同浓度的雌二醇,再室温孵育5 min,最后加入2 mol/L的NaCl,5 min后观察纳米金溶液颜色,并用紫外可见分光光度计分别测定520 nm和620 nm的吸光度值。结果表明,随着雌二醇浓度增高,纳米金溶液的颜色由红变蓝,OD620/OD520比值逐渐增加,最低可以检测到10 ng/mL的雌二醇。利用其他非特异靶标对该方法进行测试表明,结构类似物如雌三醇、雌酮、睾酮等在一定浓度下也会引起纳米金溶液变蓝,而其他类雌激素物质如己烯雌酚、双酚A和溶剂甲醇等非特异靶标即使高浓度下也不会引起纳米金溶液变蓝,表明该方法具有非常好的特异性。该研究为环境内分泌干扰物中雌二醇的快速检测提供了一种新思路。
A simple and sensitive aptamer-based colorimetric detection of 17β-estradiol was developed using unmodified gold nanoparticles(AuNPs) as colorimetric probe.It is based on the fact that aptamer can be absorbed onto the surface of AuNPs by electrostatic interaction and then the adsorbed aptamer can protect AuNPs against salt-induced aggregation.Upon the addition of 17β-estradiol,the aptamer combined with its target and dissociated from the surface of AuNPs,thus causing AuNPs aggregation under high-salt conditions and the color of such AuNPs solution changed from red to blue.In a typical experiment,the 50 μL 1 μmol/L 17β-estradiol’s aptamer solution and 50 μL AuNPs were added into microplate well and incubated for 5 min.Then 50 μL different concentrations of 17β-estradiol was added to increase a volume up to 150 μL,homogenized and allowed to stand for 5 min.Subsequently,10 μL 2 mol/L NaCl was transferred to the solution to give a final volume of 160 μL and mixed thoroughly.After the solution was equilibrated for 5 min,the UV absorbance spectrum of the solution in the range of 450-750 nm was obtained by UV/Vis spectroscopy.The results showed that,with increase of 17β-estradiol concentration,the color of the solution changed from red to blue and the ratio of OD620/OD520 was increased.As low as 10 ng/mL 17β-estradiol could be detected.In order to determine the specificity of this method,the sensing platform against several non-specific targets was tested.The structural analogues such as estriol,estrone,19-nortestosterone could also cause the color of the solution changing from red to blue with high concentration.while in the presence of other estrogen like matters(such as diethylstilbestrol,bisphenol A and the solvent of methanol),the solution would still keep red even under high concentration.The results showed that the developed method has high specificity.All of these results illustrated that the method could be readily extended toward the on-site monitoring of 17β-estradiol in environmental matrices.
适配体纳米金比色法雌二醇
aptamergold nanoparticlescolorimetry17β-estradiol
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