skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: A Fluidics-Based Biosensor to Detect and Characterize Inhibition Patterns of Organophosphate to Acetylcholinesterase in Food Materials
A chip-based electrochemical biosensor is developed herein for the detection of organophosphate (OP) in food materials. The principle of the sensing platform is based on the inhibition of dimethoate (DMT), a typical OP that specifically inhibits acetylcholinesterase (AChE) activity. Carbon nanotube-modified gold electrodes functionalized with polydiallyldimethylammonium chloride (PDDA) and oxidized nanocellulose (NC) were investigated for the sensing of OP, yielding high sensitivity. Compared with noncovalent adsorption and deposition in bovine serum albumin, bioconjugation with lysine side chain activation allowed the enzyme to be stable over three weeks at room temperature. The total amount of AChE was quantified, whose activity inhibition was highly linear with respect to DMT concentration. Increased incubation times and/or DMT concentration decreased current flow. The composite electrode showed a sensitivity 4.8-times higher than that of the bare gold electrode. The biosensor was challenged with organophosphate-spiked food samples and showed a limit of detection (LOD) of DMT at 4.1 nM, with a limit of quantification (LOQ) at 12.6 nM, in the linear range of 10 nM to 1000 nM. Such performance infers significant potential for the use of this system in the detection of organophosphates in real samples.  more » « less
Award ID(s):
1917105
PAR ID:
10339025
Author(s) / Creator(s):
; ; ; ; ; ;
Date Published:
Journal Name:
Micromachines
Volume:
12
Issue:
4
ISSN:
2072-666X
Page Range / eLocation ID:
397
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Acephate is an organophosphorus pesticide (OP) that is widely used to control insects in agricultural fields such as in vegetables and fruits. Toxic OPs can enter human and animal bodies and eventually lead to chronic or acute poisoning. However, traditional enzyme inhibition and colorimetric methods for OPs detection usually require complicated detection procedures and prolonged time and have low detection sensitivity. High-sensitivity monitoring of trace levels of acephate residues is of great significance to food safety and human health. Here, we developed a simple method for ultrasensitive quantitative detection of acephate based on the carbon quantum dot (CQD)-mediated fluorescence inner filter effect (IFE). In this method, the fluorescence from CQDs at 460 nm is quenched by 2,3-diaminophenazine (DAP) and the resulting fluorescence from DAP at 558 nm is through an IFE mechanism between CQDs and DAP, producing ratiometric responses. The ratiometric signal I 558 / I 460 was found to exhibit a linear relationship with the concentration of acephate. The detection limit of this method was 0.052 ppb, which is far lower than the standards for acephate from China and EU in food safety administration. The ratiometric fluorescence sensor was further validated by testing spiked samples of tap water and pear, indicating its great potential for sensitive detection of trace OPs in complex matrixes of real samples. 
    more » « less
  2. Oxidative stress and excessive accumulation of the superoxide (O2.-) anion are at the genesis of many pathological conditions and the onset of several diseases. The real time monitoring of (O2.-) release is important to assess the extent of oxidative stress in these conditions. Herein, we present the design, fabrication and characterization of a robust (O2.-) biosensor using a simple and straightforward procedure involving deposition of a uniform layer of L-Cysteine on a gold wire electrode to which Cytochrome C (Cyt c) was conjugated. The immobilized layers, studied using conductive Atomic Force Microscopy (c-AFM) revealed a stable and uniformly distributed redox protein on the gold surface, visualized as conductivity and surface topographical plots. The biosensor enabled detection of (O2.-) at an applied potential of 0.15 V with a sensitivity of 42.4 nA/μM and a detection limit of 2.4 nM. Utility of the biosensor was demonstrated in measurements of real time (O2.-) release in activated human blood platelets and skeletal rat limb muscles following ischemia reperfusion injury (IRI), confirming the biosensor's stability and robustness for measurements in complex biological systems. The results demonstrate the ability of these biosensors to monitor real time release of (O2.-) and estimate the extent of oxidative injury in models that could easily be translated to human pathologies. 
    more » « less
  3. The inflammation marker Interleukin 6 (IL-6) typically remains below 5 pg/mL in the serum of healthy individuals but can increase tenfold during inflammation in chronic conditions like COVID-19 and rheumatoid arthritis, as well as acute conditions like sepsis. This study is focused on the rapid detection of IL-6 to monitor both chronic and acute diseases. The novel sensor, designed with gold-coated micropyramids on the electrodes, was fabricated using the two-photon polymerization method, enabling low-volume sensing capabilities (2-3 μL). The micropyramids were surface functionalized with interleukin-6 antibodies towards developing an affinity biosensor specific to the physiological relevant range of IL-6 of 5.1 and 18.8 pg/mL in mild inflammation. Sensing was achieved by measuring impedance changes associated with IL-6 binding to the antibodies on the micropyramids interfaced using electrochemical impedance spectroscopy. It was observed that the signals from the lowest detection concentration was enhanced by 3 times at 1500 hz when the 532 nm green laser was incident on the micropyramids. This innovative approach can be expanded to the detection of cytokines not only in serum but also in respiratory samples. As a result, it opens up new avenues for monitoring local inflammation within the lungs and assessing systemic inflammation levels throughout the body. 
    more » « less
  4. Bacterial contamination in food-processing facilities is a critical issue that leads to outbreaks compromising the integrity of the food supply and public health. We developed a label-free and rapid electrochemical biosensor for Listeria monocytogenes detection using a new one-step simultaneous sonoelectrodeposition of platinum and chitosan (CHI/Pt) to create a biomimetic nanostructure that actuates under pH changes. The XPS analysis shows the effective co-deposition of chitosan and platinum on the electrode surface. This deposition was optimized to enhance the electroactive surface area by 11 times compared with a bare platinum–iridium electrode (p < 0.05). Electrochemical behavior during chitosan actuation (pH-stimulated osmotic swelling) was characterized with three different redox probes (positive, neutral, and negative charge) above and below the isoelectric point of chitosan. These results showed that using a negatively charged redox probe led to the highest electroactive surface area, corroborating previous studies of stimulus–response polymers on metal electrodes. Following this material characterization, CHI/Pt brushes were functionalized with aptamers selective for L. monocytogenes capture. These aptasensors were functional at concentrations up to 106 CFU/mL with no preconcentration nor extraneous reagent addition. Selectivity was assessed in the presence of other Gram-positive bacteria (Staphylococcus aureus) and with a food product (chicken broth). Actuation led to improved L. monocytogenes detection with a low limit of detection (33 CFU/10 mL in chicken broth). The aptasensor developed herein offers a simple fabrication procedure with only one-step deposition followed by functionalization and rapid L. monocytogenes detection, with 15 min bacteria capture and 2 min sensing. 
    more » « less
  5. Abstract The significance of easily detecting rare earth elements (REEs) has increased due to the growing demand for REEs. Addressing this need, we present an innovative electrochemical biosensor, focusing on cerium as a model REE. This biosensor utilizes a modified EF‐hand loop peptide sequence, incorporating cysteine for covalent attachment to a gold working electrode and tyrosine as an electrochemically active amino acid. The sensor was designed such that binding to cerium induces a conformational change in the peptide, affecting tyrosine's proximity to the electrode surface, modulating the current. A calibration curve was generated from cyclic voltammetry current peaks at ~0.55–0.65 V versus a silver pseudo‐reference electrode, with cerium concentrations ranging from 0 to 67 μM in artificial urine. The sensor exhibited a biologically relevant limit of detection of 35 μM and a sensitivity of −0.0024 ± 0.002 (μA μM)−1. These findings offer insights into designing peptide sequences for electrochemical biosensing. 
    more » « less