基于乙酰胆碱酯酶生物传感平台的西维因检测

doi:10.13208/j.electrochem.171229

摘要/Abstract

摘要： 为给农药西维因检测提供一种新方法，根据西维因抑制乙酰胆碱酯酶活性的原理，以黑珍珠2000（BP2000）为乙酰胆碱酯酶的固定化材料，采用滴凃电极法构建了基于乙酰胆碱酯酶的西维因生物传感平台. 结果表明，固定在BP2000 上的乙酰胆碱酯酶保持了对氯化乙酰胆碱的催化活性，并且由于BP2000 材料的引入，提升了电极有效的电化学活性表面积，而且电极上物质的电化学氧化拥有较低氧化电位（0.630 V）并伴随质子传输. 由BP2000 搭建成功的乙酰胆碱酯酶生物传感平台对西维因检测的线性响应范围为2.0 ng·mL^-1 ~ 12.5 ng·mL^-1，检测限为3.15 ng·mL^-1. 本研究对酶生物传感平台和酶生物燃料电池体系中酶电极的构建提供了一种简单方法及高效载体.

关键词: 乙酰胆碱酯酶, 西维因, 酶生物传感器, 电化学, 黑珍珠2000

Abstract:

With the purpose of providing a new method for carbaryl (a pesticide) detection, on the basis of the principle that acetylcholinesterase (AChE) activity can be restrained by carbaryl, an AChE biosensor platform based on BP2000 (as a fixation) was constructed by dropping method. As a result, it revealed that AChE immobilized on BP2000 maintained its catalytic activity for acetylcholine (ATCl), and due to the introduction of the BP2000 material, the effective electrochemical surface area of the modified electrode was enlarged. In addition, the electrochemical oxidation at the modified electrode occurred at low potential (0.630 V) accompanied by proton transmission. The AChE biosensor platform based on BP2000 matrix for carbaryl detection was able to reflect a linear response in the range of 2.0 ng·mL^-1 ~ 12.5 ng·mL^-1 with the detection limit of 3.15 ng·mL^-1. At last, this work will provide a simple method and an efficient matrix in establishing an enzyme electrode of enzymatic biosensor platform and enzymatic fuel cell.

Key words: acetylcholinesterase, carbaryl, enzyme-based biosensor, electrochemistry, black pearl 2000

中图分类号:

O646

张铭函，姜俊巧，葛君杰，邢巍. 基于乙酰胆碱酯酶生物传感平台的西维因检测[J]. 电化学（中英文）, 2018, 24(4): 303-308.

ZHANG Ming-han, JIANG Jun-qiao, Ge Jun-jie, XING Wei. Acetylcholinesterase Biosensor Platform Based on BP2000 for the Detection of Carbaryl[J]. Journal of Electrochemistry, 2018, 24(4): 303-308.

参考文献

[1] Zhang C, Cui H Y, Han Y F, et al. Development of abiomimetic enzyme-linked immunosorbent assay based on molecularly imprinted polymers on paper for the detection of carbaryl[J]. Food Chemistry, 2018, 240(1): 893-897.

[2] Liu Z M, Zang X H, Liu W H, et al. Novel method for the determination of five carbamate pesticides in water samples by dispersive liquid-liquid microextraction combined with high performance liquid chromatography[J]. Chinese Chemical Letters, 2009, 20(2): 213-216.

[3] Cheng X, Wang Q L, Zhang S, et al. Determination of four kinds of carbamate pesticides by capillary zone electrophoresis with amperometric detection at a polyamide-modified carbon paste electrode[J]. Talanta, 2007, 71(3): 1083-1087.

[4] Oh-Shin Y S, Ko M, Shin H S, et al. Simultaneous quantification of insecticides including carbaryl in drinking water by gas chromatography using dual electron-capture and nitrogen-phosphorus detection[J]. Journal of Chromatography A, 1997, 769(2): 2285-291.

[5] Patil V B, Shingare M S. Thin-layer chromatographic detection of carbaryl using phenylhydrazine hydrochloride [J].Journal of Chromatography A, 1993, 653(1): 181-183.

[6] Totti S, Fernández M, Ghini S, et al. Application of matrix solid phase dispersion to the determination of imidacloprid, carbaryl, aldicarb, and their main metabolites in honeybees by liquid chromatography-mass spectrometry detection[J]. Talanta, 2006, 69(3): 724-729.

[7] Song Y H, Chen J G, Sun M, et al. A simple electrochemical biosensor based on AuNPs/MPS/Au electrode sensing layer for monitoringcarbamate pesticides in real samples[J]. Journal of Hazardous Materials, 2016, 304(3):103-109.

[8] Kestwal R M, Bagal-Kestwal D, Chiang B, et al. Fenugreek hydrogel-agarose composite entrapped gold nano-particles for acetylcholinesterase based biosensor for carbamates detection[J]. Analytica Chimica Acta, 2015, 886(7): 143-150.

[9] Cesarino I, Moraes F C, Lanza M, et al. Electrochemical detection of carbamate pesticides in fruit and vegetables with a biosensor based on acetylcholinesterase immobilised on a composite of polyaniline-carbon nanotubes[J]. Food Chemistry, 2012, 135(3): 873-879.

[10] Liu Q, Fei A, Huan J, et al. Effective amperometric biosensor for carbaryl detection based on covalent immobilization acetylcholinesterase on multiwall carbon nanotubes/

graphene oxide nanoribbons nanostructure[J]. Journal of Electroanalytical Chemistry, 2015, 740(3): 8-13.

[11] Hatefi-Mehrjardi A. Bienzyme self-assembled monolayer on gold electrode: An amperometric biosensor for carbaryldetermination[J]. Electrochimica Acta, 2013, 114(12): 394-402.

[12] Li Y, Shi L Y, Han G Y, et al. Electrochemical biosensing of carbaryl based on acetylcholinesterase immobilized onto electrochemically inducing porous graphene oxide network[J]. Sensors and Actuators B: Chemical, 2017, 238(1): 945-953.

[13] Zhang Y, Arugula M A, Wales M, et al. A novel layer-by-layer assembled multi-enzyme/CNT biosensor for discriminative detection between organophosphorus and non-organophosphrus pesticides[J]. Biosensors and Bioelectronics, 2015, 67(5): 287-295.

[14] Zhou L Y, Zhang X, Ma L, et al. Acetylcholinesterase/chitosan-transition metal carbides nanocomposites-based biosensor for the organophosphate pesticides detection[J]. Biochemical Engineering Journal, 2017, 128(12): 243-249.

[15] Han X J, Huang W M, Jia J B, et al. Direct electrochemistry of hemoglobin in egg-phosphatidylcholine films and its catalysis to H₂O₂[J]. Biosensors and Bioelectronics, 2002, 17(9): 741-746.

[16] Han X J, Cheng W L, Zhang Z L, et al. Direct electron transfer between hemoglobin and a glassy carbon electrode facilitated by lipid-protected gold nanoparticles[J]. BBA-Bioenergetics, 2002,1556(2/3): 273-277.

[17] Wang K Q, Yang H, Zhu L, et al. Direct electron transfers and electrocatalysis of glucose oxidase immobilized on glassy carbon electrode modified with Nafion and mesoporous carbon FDU-15[J]. Electrochimica Acta, 2009, 54(20): 4626-4630.

[18] Xu X(徐璇), Lu J S(卢菊生), Liu S Q(刘松琴). Fabrication and application of cytochrome P450 electrochemcial biosensor in drug metabolism[J]. Journal of Electrochemistry(电化学), 2015, 21(1): 45-52.

[19] Chen D F, Liu Z N, Fu J Y, et al. Electrochemical acetylcholinesterase biosensor based on multi-walled carbon nanotubes/dicyclohexyl phthalate modified screen-printed electrode for detection of chlorpyrifos[J]. Journal of Electroanalytical Chemistry, 2017, 801(9): 185-191.

[20] Lu X, Tao L, Song D D, et al. Bimetallic Pd@Au nanorods based ultrasensitive acetylcholinesterase biosensor for determination of organophosphate pesticides[J]. Sensors and Actuators B: Chemical, 2018, 255(2): 2575-2581.

[21] Huang Q L, Wang Y, Lei L, et al. Photoelectrochemical biosensor for acetylcholinesterase activity study based on metal oxide semiconductor nanocomposites[J]. Journal of Electroanalytical Chemistry, 2016, 781(11): 377-382.

[22] Cui H F, Wu W W, Li M M, et al. A highly stable acetylcholinesterase biosensor based on chitosan-TiO2-graphene nanocomposites for detection of organophosphate pesticides[J]. Biosensors and Bioelectronics, 2018, 99(1): 223-229.

[23] Wang K, Tang J, Zhang Z, et al. Laccase on Black Pearl 2000 modified glassy carbon electrode: Characterization of direct electron transfer and biological sensing properties for pyrocatechol[J]. Electrochimica Acta, 2012, 70(5): 112-117.

[24] Chen D, Zhang W, Liu D, et al. Covalent coupling of organophosphorus hydrolase loaded quantum dots to carbon nanotube/Aunanocomposite for enhanced detection of methyl parathion[J]. Biosensors and Bioelectronics, 2010, 25(6): 1370-1375.

[25] Shamagsumova R V, Shurpik D N, Padnya P L, et al. Acetylcholinesterase biosensor for inhibit or measurements based on glassy carbon electrode modified with carbon black and pillar[5]arene[J]. Talanta, 2015, 144(11): 559-568.

[26] Raghu P, Kumara Swamy B E, Madhusudana Reddy T, et al. Sol-gel immobilized biosensor for the detection of organophosphorous pesticides: A voltammetric method[J]. Bioelectrochemistry, 2012, 83(2): 19-24.

[27] Li Y P, Zhang Y Y, Han G Y, et al. An acetylcholinesterase biosensor based on graphene/polyaniline composite film for detection of pesticides[J]. Chinese Journal of Chemistry, 2016, 34(1): 82-88.

[28] Caetano J, Machado S A S. Determination of carbaryl in tomato in natura using an amperometric biosensor based on the inhibition of acetylcholinesterase activity[J]. Sensors and Actuators B: Chemical, 2008, 129(1): 40-46.