构象转换型传感器对汞、 铅、 锶离子的同时检测

doi:10.7503/cjcu20180868

摘要/Abstract

摘要：

将核酸构象转换与纳米孔膜技术联用设计了一种新型高灵敏电化学传感器, 实现了对Hg ²⁺, Pb ²⁺和Sr ²⁺的分步同时检测. 使用2种分别能与Hg ²⁺及Pb ²⁺, Sr ²⁺结合的核酸适体, 将其固定在氧化铝纳米孔膜孔道内以阻碍铁氰化钾离子传导. 利用核酸适体包裹目标物时的蜷缩状态与目标物被洗脱剂洗脱后核酸适体的伸展状态之间的构象转换, 控制纳米孔通道的“开”和“关”, 使铁氰化钾溶液的氧化还原电流发生改变. 通过监测铁氰化钾溶液的电信号变化值, 可实现同时检测此3种金属离子的目的. 实验结果表明, 该传感器对3种金属离子具有很高的灵敏度和选择性, 检测的线性范围均为0.051.50 nmol/L, 对Hg ²⁺, Pb ²⁺和Sr ²⁺的检出限分别为0.013, 0.017和0.022 nmol/L(S/N=3).

关键词: 多孔阳极氧化铝, 汞离子, 铅离子, 锶离子, 生物传感器

Abstract:

A high-sensitive and specific electrochemical biosensor was designed for simultaneous detection of mercury ion(Hg ²⁺), lead ion(Pb ²⁺) and strontium ion(Sr ²⁺) by oligonucleotide conformational transformation. The biosensor was based on porous anodic alumina(PAA) modified with two oligonucleotide aptamers which could capture Hg ²⁺, Pb ²⁺ and Sr ²⁺, respectively. The aptamers can block [Fe(CN)₆] ^3-/4- ion conduction in the nanochannels of PAA. In the presence of Hg ²⁺, Pb ²⁺ and Sr ²⁺, the aptamers could bend and wrap around those metal ions to make the nanochannel unobstructed, and then, the redox current of [Fe(CN)₆] ^3-/4- could be increased. When the biosensor eluted with β-mercaptoethanol(βME), 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid(DOTA) and kryptofix 222(KP) successively, the aptamers stretched and released Hg ²⁺, Pb ²⁺ and Sr ²⁺ in sequence, leading to the re-blockage of nanochannel with the decrease of the redox current. The calibration curves of Hg ²⁺, Pb ²⁺ and Sr ²⁺ were obtained in the range of 0.05—2.50 nmol/L, and the detection limits were 0.013, 0.017 and 0.022 nmol/L, respectively. Furthermore, the biosensor is easily constructed, stable and reusable. It is hopeful to apply in the daily monitoring of harmful metal ions in drinking water or environmental water.

Key words: Porous anodic alumina, Hg ²⁺, Pb ²⁺, Sr ²⁺, Biosensor

中图分类号:

O657.1

TrendMD:

周羽婷, 汤玉娇, 邵爽, 戴诗岩, 程圭芳, 何品刚, 方禹之. 构象转换型传感器对汞、铅、锶离子的同时检测. 高等学校化学学报, 2019, 40(8): 1621.

ZHOU Yuting, TANG Yujiao, SHAO Shuang, DAI Shiyan, CHENG Guifang, HE Pingang, FANG Yuzhi. Simultaneous Detection of Mercury, Lead and Strontium Ions Based on Conformational Conversion Sensor ^†. Chem. J. Chinese Universities, 2019, 40(8): 1621.

图/表 10

Fig.1 Schematic illustration of the proposed Hg2+/Pb2+/Sr2+-biosensor

Fig.2 SEM image of the bare PAA on the biosensor surfaces

Fig.3 DPV(A) and EIS(B) curves for bare PAA(a), PAA modified with oligonucleotide H1(b), the biosensor incubated with Hg2+(c) and then eluted by βME(d) Experimental conditions: 5×10-4 mol/L K3Fe(CN)6; 1×10-7 mol/L Hg2+; 1×10-8 mol/L βME.

Fig.4 Effect of the size of PAA nanopore on the differences of the current between before and after incubation with Hg2+ Experimental conditions: 5×10-4 mol/L K3Fe(CN)6; 1×10-7 mol/L Hg2+; 1×10-8 mol/L βME.

Fig.5 DPV curves of the entire detection process for PAA/H1/H2(a), incubated with the mixture of Hg2+, Pb2+ and Sr2+(b), eluted by βME(c), DOTA(d), and KP(e) Experimental conditions: 5×10-4 mol/L K3Fe(CN)6; 1×10-7 mol/L Hg2+; 1×10-7 mol/L Pb2+ and Sr2+; 1×10-8 mol/L βME; 1×10-7 mol/L DOTA and KP.

Table 1 Elution rate constants of Hg2+, Pb2+ and Sr2+

Eluent	Ion	k/(nmol·s^-1)
βME	Hg²⁺	4.95×10^-3
	Pb²⁺	5.60×10^-4
	Sr²⁺	7.29×10^-4
DOTA	Pb²⁺	3.10×10^-3
	Sr²⁺	6.11×10^-4
KP	Sr²⁺	2.16×10^-3

Fig.6 Specificity of the biosensor for the detection of Hg2+(A), Pb2+(B) and Sr2+(C) a. Zn2+; b. Sn2+; c. Cu2+; d. Mn2+; e. Ni2+; f. Co2+; g. Cd2+; h. Ca2+; i. Mg2+; j. F-; k. Cl-; l. NO 3 - ; m. SO 4 2 - ; n. Ac-; o. P O 4 3 - ; p. CO 3 2 - ; q. S2-; r. Pb2+; s. Sr2+; t. the mixture of all the co-existent ions; u. the solution contains 10.0 nmol/L Hg2+, Pb2+ and Sr2+. Experimental conditions: 5×10-4 mol/L K3Fe(CN)6; 1×10-8 mol/L βME; 1×10-7 mol/L DOTA and KP.

Fig.7 Regeneration of the biosensor

Fig.8 Calibration curves of the biosensor for the detection of Hg2+(A), Pb2+(B) and Sr2+(C) and linear relationships between the currents and the concentration of Hg2+(A'), Pb2+(B') and Sr2+(C')

Table 2 Quantitative detection of Hg2+, Pb2+ and Sr2+ in real samples*

Sample	Contentration of M²⁺/nmol				Recovery(%)
Sample	Ion	Original	Add	Found	Recovery(%)
Tap water	Hg²⁺	Not detected	1.00	0.98	98
	Pb²⁺	Not detected	1.00	1.11	111
	Sr²⁺	Not detected	1.00	1.08	107
Drinking water	Hg²⁺	Not detected	1.00	0.90	90
	Pb²⁺	Not detected	1.00	0.93	93
	Sr²⁺	Not detected	1.00	1.03	103
River water	Hg²⁺	0.04	1.00	1.16	112
	Pb²⁺	Not detected	1.00	0.97	97
	Sr²⁺	0.16	1.00	1.11	95

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