Electrochemical Immunosensor for in Situ Detection of Brassinolide

doi:10.12133/j.smartag.SA202311001

Abstract

Abstract:

Objective Brassinolide is an important endogenous plant hormone. In this work, an electrochemical immunosensor for in situ detection of brassinolide was constructed using screen-printed electrode (SPE). Methods Au nanoparticles (AuNPs) were firstly electrodeposited on the surface of SPE electrode by electrochemical workstation, and then CuCl₂nanowires (CuCl₂ NWs) were added to the electrode, which can not only increase the conductivity of the electrode, but also Cu²⁺ can be used as a REDOX probe for the sensor. Finally, Mxene and polydopamine nanocomposite (Mxene@PDA) were selected as the modification materials for SPE electrodes because Mxene has the advantages of large surface area and good electrical conductivity, which can further amplify Cu²⁺ signals. However, Mxene is easily oxidized and unstable in air. Polydopamine (PDA) contains a large number of catechol and amino groups, which are coated on the surface of Mxene after self-polymerization by dopamine, cutting off the path of oxygen penetration, making Mxene difficult to be oxidized. Mxene@PDA can also be used as a coupling agent to fix more antibodies on the electrode surface, improving the overall biocompatibility, and improve the overall biocompatibility. Results and Discussions The sensor has a wide linear detection range: 0.1 pg/mL to 1 mg/mL, and the detection limit was 0.015 pg/ml (S/N=3). In addition, the content of endogenous brassinolide in wheat was detected by SPE electrodes in vitro and the recovery rate was 98.13% to 104.74%.While verifying the accuracy of the sensor, it also demonstrated its superior stability and sensitivity. Besides, the sensor also showed excellent application potential in the in situ detection of brassinosteroids from wheat leaves. Compared with other brassinolide detection methods, the immunosensor developed in this study has better analytical performance. Conclusions An electrochemical immunosensor for in situ detection of brassinolide was developed for the first time, providing a good electrochemical platform for in situ determination of brassinolide in plant leaves, which has great application potential in precision agriculture.

Key words: brassinolide, in situ detection, electrochemical, AuNPs, immunosensor

WEI Qian, GAO Yuanyuan, LI Aixue. Electrochemical Immunosensor for in Situ Detection of Brassinolide[J]. Smart Agriculture, 2024, 6(1): 76-88.

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Methods	Linear range	Limit of detection	Recovery	Refs
St-co-4-VP magnetic polymer beads -HPLC	10~100 µg/L	6.5 µg/L	72.3%~90.0%	［29］
SPME-LVI-HPLC	0.5~20 µg/L	0.13 µg/L	77.8%~104%	［30］
HPLC-ELSD	4.3~543.3 mg/L	1.3 mg/L	98.03%~100.53%	［7］
LC-ESI-MS	0.1~50 pmol/L	75 fmol/L	25%~32%	［31］
UHPLC-MS/MS	0.01~50 pmol/L	4.16 fmol/L	15%~96%	［32］
SPE-UHPLC-MS/MS	0.7~104 μg/kg	0.11μg/kg	84.0%~116.3%	［33］
PMME-ISD-LC-MS	0.5~1 000 pg/mL	0.1 pg/mL	82.3%~119.1%	［2］
Electrochemistry	0.1~10⁹pg/mL	0.015 pg/mL	98.13%~104.74%	This work

Samples/pg/mL	Added/pg/mL	Found/pg/mL	RSD/%	Recovery/%
0.71±0.002 8	1	1.68±0.04	2.6	98.13
	10	11.22±0.34	3.1	104.74
	100	105.42±2.95	2.8	104.67
	1 000	1 037.67±56.03	5.4	103.69

Sample	Control	100 mM NaCl
Sample	Sensor /pg/mL	Sensor/pg/mL
1	9.16	17.85
2	9.29	13.75
3	8.07	19.35
4	7.15	18.91
5	6.34	19.33
6	8.59	16.15
7	6.79	14.43
8	6.06	18.91
9	9.59	17.45
10	10.66	20.85
Average	8.17±1.54	17.70±2.29
Significance level	0.000**	0.000**