便携式黄曲霉毒素B1检测系统设计与试验

doi:10.12133/j.smartag.SA202303004

摘要/Abstract

摘要：

为实现农副产品中黄曲霉毒素B1（AFB1）的现场快速检测，设计了一款基于差分脉冲伏安法（Differential pulse voltammetry，DPV）、以STM32F103ZET6为核心处理器的便携式检测系统。系统主要包括硬件检测设备和手机App两部分，二者通过Wi-Fi通信连接。硬件检测设备主要包括DPV波形生成电路、恒电位电路及微电流检测模块；上位机App在安卓环境下开发，主要完成信号采集、数据存储等任务。应用实验室自制的AFB1传感器，在0.1 fg/ml~100 pg/ml范围内系统可以对 AFB1实现有效检测。标准溶液中的测试结果与电化学工作站CHI760e相比，最大相对误差为7.37%。对加入不同浓度AFB1的花生油样品进行检测，以CHI760e检测结果为标准，回收率为96.8%~106%；对发霉程度不同的花生样品中进行测试，与CHI760e相比，最大相对误差为7.10%。本便携式检测系统在农副产品中AFB1的现场快速检测中具有广阔的应用前景。

关键词: 黄曲霉毒素B1, 便携式系统, STM32单片机, 差分脉冲伏安法, 恒电位电路, Wi-Fi

Abstract:

To achieve rapid on-site detection of aflatoxin B1 (AFB1) in agricultural and sideline products, a portable detection system based on differential pulse voltammetry (DPV) and STM32F103ZET6 as the core processor was designed. The system consists of two main parts: hardware detection devices and a mobile App, which are connected through Wi-Fi communication. The hardware detection equipment includes a DPV waveform generation circuit, constant potential circuit, and micro current detection module. The upper computer App was developed in an Android environment and completed tasks such as signal acquisition and data storage. After completing the design, experiments were conducted to verify the accuracy of the constant potential circuit and micro current detection module. The constant potential circuit accurately applied the voltage set by the program to the electrode, with a maximum error of 4 mV. The micro current detection module converts the current into a voltage signal according to the theoretical formula and amplifies it according to the theoretical amplification factor. The laboratory-made AFB1 sensor was used to effectively detect AFB1 in the range of 0.1 fg/ml to 100 pg/ml. The maximum relative error between the test results in the standard solution and the electrochemical workstation CHI760e was 7.37%. Furthermore, peanut oil samples with different concentrations of AFB1 were tested, and the results were compared to the CHI760e detection results as the standard, with a recovery rate of 96.8%~106.0%. Peanut samples with different degrees of mold were also tested and compared with CHI760e, with a maximum relative error of 7.10%.The system's portability allows it to be easily transported to different locations for on-site testing, making it an ideal solution for testing in remote or rural areas where laboratory facilities may be limited. Furthermore, the use of a mobile App for data acquisition and storage makes it easy to track and manage testing results. In summary, this portable detection system has great potential for widespread application in the rapid on-site detection of AFB1 in agricultural and sideline products.

Key words: AFB1, portable system, STM32, differential pulse voltammetry, constant potential circuit, Wi-Fi

中图分类号:

TP216

王鹏飞, 高原源, 李爱学. 便携式黄曲霉毒素B1检测系统设计与试验[J]. 智慧农业(中英文), 2023, 5(1): 146-154.

WANG Pengfei, GAO Yuanyuan, LI Aixue. Design and Test of Portable Aflatoxin B1 Detection System[J]. Smart Agriculture, 2023, 5(1): 146-154.

图/表 16

图1

图2

图3

图4

图5

图6

图7

图8

表1

表2

表3

图9

表4

表5

图10

表6

参考文献 32

1	SINGH A K, DHIMAN T K, V S L G B, et al. Dimanganese trioxide (Mn₂O₃) based label-free electrochemical biosensor for detection of aflatoxin-B1[J]. Bioelectrochemistry (amsterdam, Netherlands), 2021, 137: ID 107684.
2	WANG C J, ZHANG L, LUO J Y, et al. Development of a sensitive indirect competitive enzyme-linked immunosorbent assay for high-throughput detection and risk assessment of aflatoxin B₁ in animal-derived medicines[J]. Toxicon: Official journal of the International Society on Toxinology, 2021, 197: 99-105.
3	JAHANGIRI-DEHAGHANI F, ZARE H R, SHEKARI Z, et al. Development of an electrochemical aptasensor based on Au nanoparticles decorated on metal-organic framework nanosheets and p-biphenol electroactive label for the measurement of aflatoxin B1 in a rice flour sample[J]. Analytical and bioanalytical chemistry, 2022, 414(5): 1973-1985.
4	FENG B B, YOU J, ZHAO F, et al. A ratiometric fluorescent aptamer homogeneous biosensor based on hairpin structure aptamer for AFB1 detection[J]. Journal of fluorescence, 2022, 32(5): 1695-1701.
5	LI Q, LI Y, GAO Q, et al. Real-time monitoring of isothermal nucleic acid amplification on a smartphone by using a portable electrochemical device for home-testing of SARS-CoV-2[J]. Analytica chimica acta, 2022, 1229: ID 340343.
6	SHI L, WANG Z F, YANG G M, et al. A novel electrochemical immunosensor for aflatoxin B1 based on Au nanoparticles-poly 4-aminobenzoic acid supported graphene[J]. Applied surface science, 2020, 527: ID 146934.
7	AKGÖNÜLLÜ S, YAVUZ H, DENIZLI A. SPR nanosensor based on molecularly imprinted polymer film with gold nanoparticles for sensitive detection of aflatoxin B1[J]. Talanta, 2020, 219: ID 121219.
8	GELETA G S, ZHAO Z, WANG Z. A novel reduced graphene oxide/molybdenum disulfide/polyaniline nanocomposite-based electrochemical aptasensor for detection of aflatoxin B1[J]. Analyst, 2018, 143: 1644-1649.
9	ZHENG W L, TENG J, CHENG L, et al. Hetero-enzyme-based two-round signal amplification strategy for trace detection of aflatoxin B1 using an electrochemical aptasensor[J]. Biosensors & bioelectronics, 2016, 80: 574-581.
10	WANG P F, LUO B, LIU K, et al. A novel COOH-GO-COOH-MWNT/pDA/AuNPs based electrochemical aptasensor for detection of AFB1[J]. RSC Advances, 2022, 12(43): 27940-27947.
11	王文廉, 牛博怀, 王玉. 用于智能手机的无源POCT尿酸检测系统设计[J]. 传感技术学报, 2021, 34(3): 413-419.
	WANG W L, NIU B H, WANG Y. Design of passive POCT uric acid detection system based on smart phone[J]. Chinese journal of sensors and actuators, 2021, 34(3): 413-419.
12	管海翔, 陈娟, 祁欣. 基于高灵敏度电化学传感器的有害气体检测系统设计[J]. 北京化工大学学报(自然科学版), 2020, 47(2): 107-114.
	GUAN H X, CHEN J, QI X. A measurement system for harmful gases based on a high sensitivity electrochemical sensor[J]. Journal of Beijing university of chemical technology (natural science edition), 2020, 47(2): 107-114.
13	ZHANG R, ZHANG J, TAN F, et al. Multi-channel AgNWs-doped interdigitated organic electrochemical transistors enable sputum-based device towards noninvasive and portable diagnosis of lung cancer[J]. Materials today bio, 2022, 16: ID 100385.
14	XIA S Q, PAN J F, DAI D S, et al. Design of portable electrochemiluminescence sensing systems for point-of-care-testing applications[J]. Chinese chemical letters, 2023, 34(5): ID 107799.
15	DU Y X, MO Z L, WANG J A, et al. A novel chiral carbon nanocomposite based on cellulose gum modifying chiral tri-electrode system for the enantiorecognition of tryptophan[J]. Journal of electroanalytical chemistry, 2021, 895: ID 115390.
16	LIU Y B, HUANG Z, XU Q, et al. Portable electrochemical micro-workstation platform for simultaneous detection of multiple Alzheimer's disease biomarkers[J]. Microchimica acta, 2022, 189(3): ID 91.
17	郭文川, 李思睿, 杨烨, 等. 基于LED的便携式牛乳亚硝酸盐含量检测仪研究[J]. 农业机械学报, 2022, 53(10): 379-385.
	GUO W C, LI S R, YANG Y, et al. Study on portable nitrite content detector in milk based on LED[J]. Transactions of the Chinese society for agricultural machinery, 2022, 53(10): 379-385.
18	甄俊杰, 曾令文. 便携式电化学粮食重金属离子检测仪的研制[J]. 现代食品, 2022, 28(11): 97-101.
	ZHEN J J, ZENG L W. Development of a portable electrochemical detector for grain heavy metal ions[J]. Modern food, 2022, 28(11): 97-101.
19	周兴辉, 胡敬芳, 宋钰, 等. 基于树莓派的便携式水质重金属电化学检测系统[J]. 现代电子技术, 2021, 44(23): 134-137.
	ZHOU X H, HU J F, SONG Y, et al. Portable water quality heavy metal electrochemical detection system based on Raspberry Pi[J]. Modern electronics technique, 2021, 44(23): 134-137.
20	SCOTT A, SAKIB S, SAHA S, et al. A portable and smartphone-operated photoelectrochemical reader for point-of-care biosensing[J]. Electrochimica acta, 2022, 419: ID 140347.
21	CORDOVA-HUAMAN A V, JAUJA-CCANA V R, LA ROSA-TORO A. Low-cost smartphone-controlled potentiostat based on Arduino for teaching electrochemistry fundamentals and applications[J]. Heliyon, 2021, 7(2): ID e06259.
22	HAO N, HUA R, ZHANG K, et al. A sunlight powered portable photoelectrochemical biosensor based on a potentiometric resolve ratiometric principle[J]. Analytical chemistry, 2018, 90(22): 13207-13211.
23	ZHOU Z Z, WANG J, LI G H, et al. Wireless USB-like electrochemical platform for individual electrochemical sensing in microdroplets[J]. Analytica chimica acta, 2022, 1197: ID 339526.
24	WANG C M, HSIEH C H, CHEN C Y, et al. Low-voltage driven portable paper bipolar electrode-supported electrochemical sensing device[J]. Analytica chimica acta, 2018, 1015: 1-7.
25	KHETANI S, SINGH A, BESLER B, et al. μDrop: Multi-analyte portable electrochemical-sensing device for blood-based detection of cleaved tau and neuron filament light in traumatic brain injury patients[J]. Biosensors & bioelectronics, 2021, 178: ID 113033.
26	邹绍维, 燕朝果, 梁东江, 等. 基于模糊PID控制的阴极保护恒电位仪设计[J]. 机电工程技术, 2021, 50(11): 243-246.
	ZOU S W, YAN C G, LIANG D J, et al. Design of cathodic protection potentiostat based on fuzzy PID control[J]. Mechanical & electrical engineering technology, 2021, 50(11): 243-246.
27	郭志涛, 杨文乐, 卢成钢, 等. 数字恒电位仪远程自动化监控系统设计[J]. 单片机与嵌入式系统应用, 2020, 20(7): 32-36.
	GUO Z T, YANG W L, LU C G, et al. Design of digital potentiostat remote automation monitoring system[J]. Microcontrollers & embedded systems, 2020, 20(7): 32-36.
28	程楚皓, 林伟国. 便携式水体重金属离子浓度快速检测仪设计[J]. 电子测量技术, 2019, 42(2): 112-116.
	CHENG C H, LIN W G. Design of portable instrument forrapid detection of heavy metal ion concentration in water[J]. Electronic measurement technology, 2019, 42(2): 112-116.
29	ZHONG T T, LI S S, LI X, et al. A label-free electrochemical aptasensor based on AuNPs-loaded zeolitic imidazolate framework-8 for sensitive determination of aflatoxin B1[J]. Food chemistry, 2022, 384: ID 132495.
30	PIERINI G D, MACCIO S A, ROBLEDO S N, et al. Screen-printed electrochemical-based sensor for taxifolin determination in edible peanut oils[J]. Microchemical journal, 2020, 159: ID 105442.
31	LI Y Y, LIU D, ZHU C X, et al. Sensitivity programmable ratiometric electrochemical aptasensor based on signal engineering for the detection of aflatoxin B1 in peanut[J]. Journal of hazardous materials, 2020, 387: ID 122001.
32	MACIEL-SILVA F W, LACHOS-PEREZ D, BULLER L S, et al. Green extraction processes for complex samples from vegetable matrices coupled with on-line detection system: A critical review[J]. Molecules, 2022, 27(19): ID 6272.

R_RW/kΩ	设定电压值/V
R_RW/kΩ	-2.0	-1.0	0	1.0	2.0
1	-1.998	-0.999	0.001	1.003	2.001
10	-1.996	-0.998	0.002	1.004	2.002
100	-1.998	-0.999	0.000	1.002	2.001
1000	-1.997	-0.996	0.001	1.004	2.000

I_输入/μA	V_理论/V	V_实测/V	绝对误差/V	相对误差/%
10	0.491	0.479	0.012	2.523
20	0.983	1.010	0.026	2.653
40	1.966	2.003	0.036	1.841
60	2.949	2.924	0.025	0.861
90	4.424	4.479	0.054	1.216

浓度/（fg·ml^-1）	CHI760e电流/μA	系统电流/μA	相对误差/%
0	80.34±0.19	80.24±0.23	1.3
0.1	72.71±0.21	69.56±0.22	4.3
100.0	40.66±0.18	42.00±0.24	3.2
100，000.0	3.05±0.17	2.99±0.19	1.8

浓度/（fg·ml^-1）	CHI760e/（fg·ml^-1）	便携式系统/（fg·ml^-1）	相对误差/%
0	0.00±0.00	0.00±0.00	——
1	0.98±0.40	0.91±0.05	7.02
10	10.51±0.03	11.27±0.02	6.72
100	97.30±0.05	90.12±0.06	7.37
1000	989.70±0.02	921.31±0.02	6.91

加入的AFB1浓度/（fg·ml^-1）	实际检测AFB1浓度/（fg·ml^-1）	相对标准偏差/（RSD，%）	回收率/%
0.0	0.00	0.00	100.0
12.5	12.10±0.22	7.04	96.8
50.0	53.20±0.43	12.00	106.0
200.0	206.40±0.95	8.01	103.2