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Electrochemical Sensor for Antibiotic Detection

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Rafia Masood, Abdur Rahim, Abdul Wajid, Sana Sabahat, Zia Ul Haq Khan, Abdul Qadeer and Hafiz Irshadullah

Submitted: 13 September 2024 Reviewed: 21 September 2024 Published: 19 December 2024

DOI: 10.5772/intechopen.1008295

Current Developments in Biosensor Applications and Smart Strategies IntechOpen
Current Developments in Biosensor Applications and Smart Strategi... Edited by Selcan Karakuş

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Current Developments in Biosensor Applications and Smart Strategies [Working Title]

Associate Prof. Selcan Karakuş

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Abstract

The development and uses of electrochemical biosensors for antibiotic detection are covered in detail in this chapter. Electrochemical biosensors have emerged as a possible answer to the pressing demand for quick and sensitive detection techniques due to the growing prevalence of antibiotic resistance. The first section introduces the fundamental concepts of antibiotics and their electrochemical sensing, including the mechanisms of electrochemical transduction and the various types of electrodes and transducers employed. Next, several biosensor designs, including those based on enzymes, aptamers, and molecularly imprinted polymers, are examined for antibiotic detection. Key performance measures are reviewed, along with obstacles and constraints in the field, including sensitivity, specificity, and reaction time. The chapter emphasizes the combination of nanomaterials and microfluidics to improve biosensor functioning, as well as current advancements and future directions. It aims to provide valuable insights to researchers and practitioners in the field of electrochemical biosensing for antibiotic detection by offering a comprehensive overview of current technologies and developments.

Keywords

  • antibiotics
  • electrochemical sensors
  • life-saving drugs
  • nanomaterials
  • biosensors

1. Introduction

In today’s healthcare system, antibiotics are a class of antimicrobial medications that are extensively utilized to treat bacterial infections. In the medical field, antibiotics have become revolutionary therapies for a variety of infectious diseases that affect both humans and animals. Antibiotics are chemotherapeutic medicines that inhibit or stop the growth of bacteria; they are referred to be “wonder drugs” [1]. Nevertheless, certain bacteria have developed resistance to particular antibiotics as a result of overuse. Because of their structural complexity, the majority of antibiotics pass through urine and feces unchanged, damaging natural water sources and soil in the process [2].

Lately, the widespread use of antibiotics in animal feed to boost development and productivity has also tainted food items utilized by humans [3].

Antibiotic residues, however, may be highly hazardous if they are present in large enough quantities. They can lead to infections such as gonorrhea, pneumonia, and tuberculosis (TB), which can complicate treatment. Antibiotic resistance has been associated with humans not only with infections but also with severe allergy responses. According to WHO (World Health Organization) Guidelines (2015) [4], 300 million premature deaths are predicted to occur globally over the next 35 years if the current trend continues.

Most of the time, antibiotics prevent infections or germs by blocking the creation of proteins, folic acid, DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and bacterial cell walls [5].

To specifically affect bacterial and microbe populations in a dosage-dependent way, antibiotics have been created. According to Alanis [6], the deleterious impact of antibiotics causes microbial populations to vanish or be eliminated due to their bacteriostatic and bactericidal properties.

Antibiotic dosage modification and trace-level monitoring are particularly desirable to reduce the emergence of antibiotic-resistant bacteria. Numerous analytical techniques like chromatography, mass spectrometry, solid-phase microextraction, capillary electrophoresis, enzyme-linked immunosorbent assay, surface plasmon resonance, etc., have been created to date for precise and trustworthy monitoring of antibiotic residues in natural resources. Electrochemical biosensors are a portable, reliable, and fast-response diagnostic tool for high-sensitivity antibiotic residue probing when compared to current analytical approaches [7].

A biosensor is a diagnostic device that utilizes immobilized biological components, such as antibodies, antigens, hormones, and enzymes, or the organism’s sensitive elements, such as cells, tissues, and organelles. Biosensors are highly selective due to the use of biologically sensitive elements, making them a perfect analytical instrument for swiftly and directly getting complicated system compositional data [8]. In recent years, A variety of modification components have been developed to address the demand for detection under varied, complicated situations, including aptamers, nanomaterials, antibodies, and detecting techniques. Novel biosensors can profit greatly from the introduction of biological materials, which can improve the stability and specificity of target-detecting compounds [9].

The deployment of electrochemical biosensors has shown great promise in the identification of antibiotics because of their high sensitivity, selectivity, and ability to conduct real-time, quick analysis [9].

Recent advancements have placed significant emphasis on integrating polymers and nanomaterials into the development of electrochemical biosensors, aiming to leverage these materials for boosting sensor sensitivity and detection limits [10, 11]. Graphene, carbon nanotubes, and metal nanoparticles, among other polymers and nanomaterials, have found application in enhancing the stability and overall performance of biosensors through the immobilization of receptors [10].

Moreover, there has been an increasing focus on creating non-enzymatic biosensors that utilize synthetic materials instead of proteins. This is due to the potential for these materials to provide enhanced electrode stability and selectivity, as highlighted by Vigneshvar et al. in 2016 [12].

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2. Antibiotics and their types

Antibiotics, a secondary metabolite, are produced by microorganisms, higher animals, and plants and possess antipathogenic properties. These substances can hinder the growth of other cells and are widely employed in veterinary and human medicine, agriculture, animal husbandry, aquaculture, and various other fields [13]. Their usage is focused on the treatment and prevention of bacterial infections [14]. Regrettably, improper use of antibiotics in humans can lead to ototoxicity, nephrotoxicity, and antibiotic resistance [15, 16]. Different antibiotics are used to treat other types of bacteria because each bacterium has a specific antibiotic to combat it. Most of the various kinds of antibiotics are categorized according to their chemical makeup, mode of action, range of action, and mode of administration. These seven groups of antibiotics: phenicol, sulfonamides (SAs), tetracyclines (TCs), aminoglycosides (AGSs), β-lactams (BLCs), fluoroquinolones (FQs), and macrolides are the most often administered antibiotics in clinics [17].

2.1 Phenicol

Phenicol, a type of antibiotic, is employed in the treatment of specific bacterial infections. They function by stopping bacteria from producing essential proteins that they need to live, thereby reducing the growth of the bacteria. Chloramphenicol (CAP) and thiamphenicol (TAP), both belonging to the phoenix group, can be seen in Figure 1 due to their similar chemical structures. CAP is widely used in human and animal healthcare because it is cost-effective and potent. Yet, its overuse encourages bacterial resistance, leading to its decreased effectiveness in treating diseases. Furthermore, this overuse has caused increased amounts of CAP residues in animal-derived products like milk, meat, eggs, and liver [18, 19, 20]. Thus, residual CAP present in these products can be transferred to humans, causing a range of negative side effects such as diarrhea, allergic responses, suppression of bone marrow, nausea, gray baby syndrome, and damage to the kidneys [21].

Figure 1.

Chemical structure of chloramphenicol (CAP) and thiamphenicol (TAP) [the figures are redrawn by the authors].

2.2 Sulfonamides (SAs)

Sterile antibiotics, also known as SAs, are synthetic antibiotics utilized in chemotherapy. While some SAs possess anti-bacterial capabilities, others serve additional medical purposes, like inducing diuresis, treating glaucoma, or managing seizures. SAs play a role in folic acid synthesis, thereby depriving bacteria of this essential vitamin. The use of SAs to reduce their negative clinical effects can be enhanced by incorporating additional chemical groups. These compounds are marketed under various brand names such as sulfadimidine, sulfadimethoxine (SDM), sulfamonomethoxine, and sulfadiazine (SD). Due to their reliable and extensive ability to fight bacteria, as well as their low cost, synthetic amino acids (SAs) are widely employed in human healthcare as well as in the fields of aquaculture and animal breeding. SAs can end up in the environment through human and animal waste, which can pollute water sources. Exposure to lingering SAs in the body can lead to various health issues, including kidney stone formation, thyroid cancer, and severe allergic reactions [22, 23].

2.3 Tetracyclines (TCs)

The TCs extracted from a specific type of Streptomyces bacteria consisted of four rings, as shown by the chemical structure in Figure 2. TCs are widely used antibiotics that function by stopping bacteria from producing proteins. Because of their various antibacterial qualities, remarkable therapeutic effects, and affordability, TCs have been used widely in the treatment of infections in both people and animals [24, 25]. Unfortunately, the amount of residual pesticides (TCs) in plants, drinking water, animal food items, and the environment has increased as a result of the extensive and uncontrolled use of TCs (Figure 2) [26].

Figure 2.

Chemical structure of tetracyclines (TCs) [The figures are redrawn by the authors].

2.4 Aminoglycosides (AGSs)

AGSs are derived from many species of Streptomyces and share the streptamine ring Figure 3 as their common core structure. By preventing Gram-positive and Gram-negative bacteria from generating proteins, AGSs work as bactericidal drugs to treat bacterial infections. The use of AGSs in people and animals led to an increase in bacterial resistance. As a result, to combat germs with two different approaches, AGSs are employed in tandem with PENs or CAPs. AGSs have been used less often in clinical settings due to their negative side effects, especially their ototoxicity and nephrotoxicity [27]. On the other hand, because of their low cost, they are nevertheless frequently and widely used as a food supplement in animal production, which probably results in residues in the food products that come from animals [28].

Figure 3.

Chemical structures of representative aminoglycosides, highlighting the atypical compounds streptomycin and apramycin. [The figures are redrawn by the authors].

2.5 β-Lactams (BLCs)

PENs and cephalosporins (CEPHs) are the most prevalent forms of BLCs. The chemical substance known as penicillin, or PENs, has the formula R-C9H11N2O4S, with R varying depending on the kind of penicillin. The mold Penicillium notatum generates inexpensive PENs that prevent bacteria from developing cell walls. PENs are used to treat infections of the respiratory system, ear, skin, and dental cavities, as well as gonorrhea [29, 30]. CEPHs are produced by the plant Cephalosporin, and while they have a mode of action similar to PENs, they have distinct underlying chemical structures Figure 4. CEPHs, like PENs, have a β-lactam ring structure that inhibits the synthesis of bacterial cell walls. CEPHs effectively cure a wide range of bacterial infections, including tonsillitis, otitis media, bronchitis, pneumonia, and strep throat [31].

Figure 4.

Basic structure of (a) PENs and (b) CEPH [The figures are redrawn by the authors].

2.6 Fluoroquinolones (FQs)

FQs, as the name implies, are synthetic antibiotics derived from the root “floxacin.” FQs were recently discovered to have a broad antibiotic spectrum and can cure respiratory, cutaneous, and urinary tract infections. FQs block bacteria from producing DNA, hence suppressing their growth. Figure 5 depicts the chemical structures of norfloxacin (NOR) and enrofloxacin (ENRO). NOR can only be used to treat people, but ENRO was designed particularly for veterinary usage.

Figure 5.

Basic structure of (a) ENOR and (b) NOR (R1 = H, R2 = C2H5) [The figures are redrawn by the authors].

FQs are synthetic antibiotics derived from the root “floxacin,” as indicated by their common name. Recently, it was discovered that FQs can treat infections of the respiratory, cutaneous, and urinary tracts and have a wide antibiotic range. FQs stop bacteria from making DNA and stop them from growing [32, 33]. The synthetic designs of Norfloxacin (NOR) and enrofloxacin (ENRO) are portrayed in Figure 6. NOR must be utilized for treating people, while ENRO was made explicitly for veterinary purposes.

Figure 6.

Basic structure of azithromycin (AZM) [The figures are redrawn by the authors].

2.7 Macrolides

The macrolide anti-microbials were separated utilizing Streptomyces microscopic organisms, which have a macrocyclic lactone structure. Macrolides resemble PENs in their broadness of purpose and are bacteriostatic medications that keep bacterial ribosomes from combining proteins. The most broadly utilized macrolides for treating gastrointestinal lot, cutaneous, vaginal, and respiratory parcel contaminations incorporate roxithromycin, erythromycin, clarithromycin (CIP), and azithromycin (AZM) as displayed in Figure 6.

Antimicrobial resistance has been firmly associated with the abuse and abuse of anti-toxins in agribusiness, as well as the resulting ecological defilement (AMR). The well-established pecking order is likewise adversely affected by this development. A wide range of antibiotics, including lactams, aminoglycosides, fluoroquinolones, and sulphonamides, have been listed as potential environmental pollutants that could harm public health by the World Health Organization. Anti-infection pollution is on the ascent, and this could have unsafe ramifications for creatures, plants, microorganisms, and, at last, individuals. Antibiotic-resistant bacteria, which are also considered a new class of environmental contaminants, evolved through genetic or mutational changes as a result of the widespread use of antibiotics. Anti-toxins are currently present basically wherever in the climate, including soils and water assets, due to their unhindered utilization. Anti-toxin deposits might be available in wastewater and other natural squanders from emergency clinics, cultivating soils, and the drug business. If these squanders are delivered into the climate untreated, they might blend in with water assets, as displayed in Figure 6. Anti-microbial safe microorganisms (ARB) are remembered to flourish in clinic garbage. Contrasted with wild strains, ARB strains can persevere and multiply in serious conditions [17].

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3. Methods of detection of antibiotic

Antibiotics are generally utilized in the veterinary business and hydroponics, and the abuse and mistaken utilization of anti-microbials will prompt natural contamination and undermine human well-being. To address or mitigate the aforementioned issues, it is necessary to investigate sensitive and effective antibiotics in wastewater detection techniques. This paper audits the better discovery techniques for anti-infection agents in wastewater in the past 5 years, which is helpful for the subsequent investigation of more basic, touchy, and precise recognition strategies [1].

3.1 Pretreatment method

Aquaculture effluent may have barely any residual antibiotics. To enhance and refine antibiotics for detection, it is often pretreated before determining the presence of antibiotic content. Magnetic solid-phase extraction, solid-phase extraction, and solid-phase microextraction are the key topics covered in this chapter [34].

3.1.1 Solid-phase extraction (SPE)

Solid-phase extraction column fitted with high-efficiency extraction fillers is the primary means of concentrating, separating, and purifying target materials in water samples; Figure 7 illustrates the general flow of the SPE. Adsorbents come in a variety of forms, including lignocellulosic materials and ionic liquids [35], graphene oxide composite microspheres [36], covalent organic framework (COF) [37], MOF, and so forth. Solid-phase extraction (SPE) has gained significant attention in the extraction field due to its simplicity, cost-effectiveness, high selectivity, and reliable recovery rates [38].

Figure 7.

Schematic representation of SPE. [The figures are redrawn by the authors].

The combined effect of various adsorbent materials enhanced the adsorption capacity of the target substance, achieving a high recovery rate through high-performance liquid chromatography [34].

3.1.2 Solid-phase microextraction (SPME)

One-step simultaneous extraction and preconcentration of materials is possible with SPME, a multifunctional sample preparation method [39]. For thin-film SPME, on-fiber SPME, in-tube SPME, and stir bar SPME, among others, there are many extraction devices [40]. The SPME has been employed in numerous research as a pretreatment approach for antibiotic detection.

To identify perfluoro alkylated aromatic hydrocarbons (PFAAs) in water samples, Gong et al. [41] synthesized a wrapping material for solid-phase microextraction fibers, amino-functionalized ZIF-8 (NH-ZIF-8). High-performance liquid chromatography (HPLC)–Mass spectrometry (MS)/MS was then used to identify PFAAs in water samples. The linear range of this approach is 1–5000 ng/L, with an identification limit of 0.15–0.75 ng/L. It has been effectively used for environmental water analysis. In the extraction phase of SPME, Wu et al. [42] developed a monolith/aminated graphene oxide composite material on stainless-steel wire, which was coupled with a DC power supply to enable electric field-assisted SPME technology. Their results demonstrated that the electric field improved the enrichment of trace phenoxy carboxylic acid herbicides in environmental fluids, achieving detection limits of 0.54–1.3 μg/L within a linear range of 0.005–50.0 μg/L.

3.1.3 Magnetic solid-phase extraction (MSPE)

The increasing prevalence of magnetic adsorbents, a relatively new technology, has led to the development of SPE technologies such as MSPE Figure 8. For certain residues, magnetic adsorbents can adsorb materials quite well [43].

Figure 8.

Magnetic solid-phase extraction (MSPE). [The figures are redrawn by the authors].

Future development research will focus heavily on the creation of ecologically friendly materials. Furthermore, technology for imprinting has gradually developed. He et al. [44] employed a surface imprinting method to develop an entirely novel nanocomposite Fe3O4@SiO2 MMIP with magnetic response, which they then used for the determination of doxycycline in marine sediments by HPLC. The method has good linearity, high sensitivity, and low environmental impact when compared to the currently used methods. The most notable aspect of this study is that the environmentally friendly synthesis of maltodextrin nanosponges (MDNSs) as adsorbents inspires the consequent development of sustainable products [34].

3.2 Techniques for antibiotic detection

3.2.1 Immunoassay

The immunoassay technique leverages the specific interaction between an antigen and its corresponding antibody, providing advantages such as a low detection limit and high selectivity. Although much of the research on antibiotic detection in wastewater has been centered on identifying antibiotics and antibiotic-related compounds in food and the environment, Wang et al. [45] developed an innovative immunoassay using an avidin-biotin complex. This approach involves biotin-labeled up conversion nanoparticles combined with streptavidin to form an avidin-biotin upconversion complex, which then binds to biotinylated antibodies, thereby enhancing the detection signals. This method exhibits high sensitivity and high flux, with detection limits of 0.05, 0.05, and 0.15 ng/mL, respectively.

3.3 Instrumental analysis method

One of the most used techniques for identifying antibiotics these days is instrumental analysis. Spectrophotometry, mass spectrometry, and chromatography are the three categories of instrumental analytical techniques.

3.3.1 Chromatography

High-performance liquid chromatography (HPLC) which is the most prevalent chromatography technique for nonvolatile analytes [46]. High-performance liquid chromatography (HPLC) is employed to isolate specific compounds from mixtures by leveraging the interactions between the stationary phase within the column and the mobile phase. Depending on the analyte, various adsorption principles are utilized for effective separation and elution [47]. There is a fair amount of research on HPLC in the field of antibiotic detection, and most studies are currently not restricted to a single identification method; instead, HPLC–tandem mass spectrometry is typically employed for antibiotic detection. Ultra-trace FQs were identified and quantified by Yu et al. [48] in wastewater by using techniques of molecularly imprinted SPE and liquid chromatography–tandem mass spectrometry.

Zhu et al. [44] identified environmental water for trace quinolone antibiotics by using an isotope dilution UHPLC-tandem mass spectrometry and solid-phase extraction. The linear range 0.05–100 μg/L was for 15 quinolones, with detection limits and correlation coefficients ranging from 0.005 to 0.051 ng/L and 0.9993–0.9999, respectively.

3.3.2 Mass spectrometry (MS)

By employing an electric field and magnetic field to separate the ions based on their mass-to-charge ratio, mass spectrometry (MS) is a technique for detecting moving ions; it can be further classified into four types: time-of-flight mass spectrometry, quadrupole mass spectrometry, Orbitrap mass spectrometry, and magnetic sector mass spectrometry [45]. Typically, MS is used in conjunction with chromatography or spectroscopy to detect antibiotics in wastewater; for instance, Krakkó et al. [49] employed solid-phase preconcentration to extract samples from drinking water, river water, and treated wastewater. These extracts were analyzed using ultra-high-performance liquid chromatography (UHPLC), followed by quadrupole time-of-flight mass spectrometry, to quantitatively determine ten commonly used pharmaceutical active ingredients.

3.3.3 Spectroscopy

Spectroscopy allows a substance to either emit or absorb photon energy to find antibiotic residues. By measuring the sample’s absorbance or emission light intensity at various wavelengths, this approach primarily evaluates the composition and concentration of chemicals in the sample. Antibiotics in wastewater are often detected using ultraviolet–visible (UV–Vis) spectroscopy and surface-enhanced Raman spectroscopy (SERS), the latter of which has been the subject of several investigations.

Antibiotics in wastewater may be effectively monitored in real-time using UV–Vis spectroscopy. Very few studies have directly employed this approach to determine antibiotics; instead, it is often used to define the antibiotics photodegradation catalyst. Using a UV–visible sensor, Li et al. [50] determined the absorption spectra of wastewater samples containing various amounts of ofloxacin, tetracycline, and chloramphenicol. The findings demonstrated that the three antibiotics’ average detection limit was 0.85 mg/L at a sensor optical path length of 0.5 mm. Ofloxacin absorption peak at 290 nm indicated that 0.4647 mg/L was the minimal detection limit.

Antibiotics were detected down to 3.1 μg/L at the minimum when the sensor’s optical path length was 10 cm. Large mistakes will be produced when UV–Vis spectroscopy is used directly due to the instrument’s resolution and range limits. Therefore, to make up for its limitations, new technologies must be developed. To find antibiotics in bodily fluids, Domes et al. [51] investigated deep UV resonance Raman spectroscopy. The benefits of deep UV resonance Raman spectroscopy include excellent sensitivity, quick processing times, and the ability to perform multi-component determination using multivariate identification. Multiple antibiotics in complicated samples may be detectable with high sensitivity if this is investigated.

3.3.4 Sensors

Antibiotic sensor detection is currently one of the most investigated and extensive ways; typically used sensors comprise surface plasmon resonance (SPR), optical sensors, photoelectrochemical (PEC) sensors, biosensors, and electrochemical sensors [34].

3.3.4.1 Electrochemical sensor

3.3.4.1.1 Electrochemical detection of antibiotics

Electrochemical sensors have demonstrated great promise in the last few years for the detection of antibiotic residues because of their many advantages, including high sensitivity, quick analysis times, and the ability to be miniaturized through the use of combinations of potential (E), current (I), charge (Q), and time (t) [52]. The most often used voltammetric methods for electrochemical detection of antibiotic residues to date are stripping voltammetry, cyclic voltammetry (CV), differential pulse voltammetry (DPV), and square wave voltammetry (SWV). As an alternate method for detecting antibiotics, chronoamperometry and electrochemical impedance spectroscopy have also attracted a lot of interest.

3.3.4.1.2 Components and working principles of electrochemical sensors

A reference electrode, a counter or auxiliary electrode, and a working electrode—also referred to as the sensing or redox electrode—are typically needed for electrochemical sensing, as shown in Figure 9. To keep the reference electrode, which is typically constructed of Ag/AgCl, at a safe distance from the reaction site. The counter electrode connects to the electrolytic solution to enable the application of current to the working electrode, which acts as the transducer component in the biological reaction. These electrodes ought to be chemically stable and conductive. As a result, depending on the analyte, platinum, gold, carbon (such as graphite), and silicon compounds are frequently utilized [53].

Figure 9.

Working of electrochemical sensor for antibiotic detection [The figures are redrawn by the authors].

Changes in current, potential, charge, impedance, or conductivity are the outcome of an electrochemical reaction in this system. Depending on the parameters, these changes can be detected using a variety of techniques [54]. Voltammetry, electrochemical impedance spectroscopy (EIS), and chronoamperometry (I-t) are examples of electroanalytical techniques for the electrochemical detection of antibiotics [53]. Four voltammetry methods are frequently used to identify antibiotics [55]: (i) CV, which is useful for researching reversible redox reactions and may simultaneously observe the redox peak potential and current. (ii) SWV, which is more sensitive than CV and uses a fast-scanning stepped voltage on the electrode. (iii) DPV, whose sensitivity is likewise rather high, and which can offer pertinent analytes information on the chemical form. (iv) Linear sweep voltammetry (LSV), a technique for applying an electrode with a voltage that changes linearly. EIS can be used to evaluate electrode function and is a reliable way to identify antibiotics [56]. A test chemical performs an oxidation–reduction reaction at a constant potential while being detected by the I-t technique, which is often used to identify antibiotics [57].

Electrodes themselves are essential to the operation of electrochemical sensors since reactions are typically only perceived near the surface of electrode. Based on the electrode’s selected function, the material, the surface modification, or the electrode’s dimensions have a notable impact on how well it can detect objects. The constructed electrodes-based sensor devices have a wide linear response spectrum, high repeatability, and the capacity to detect at low concentration limits. To identify minute levels of electroactive substances in medicines and other chemical ingredients, electrochemical sensors generally employ a variety of electrochemical techniques. Since transducers are used to turn the measured quantity into an electrical signal, electrochemical sensors are crucial because they can identify even minute levels of pollutants [58].

3.4 Approaches for electrochemical detection of antibiotic contaminants

3.4.1 Electrochemical biosensor mediated by receptors or enzymes

Electrochemical biosensors have rendered it possible to identify a variety of antibiotic forms by utilizing several receptors or enzymes that have been tagged with nanomaterials. Most of these studies successfully employed carbon-based nanomaterials, colloidal gold tags, magnetic nanoparticles, and various enzymes—such as penicillinase (PCN), glucose oxidase (GOx), and horseradish peroxidase (HRP)—for the electrochemical detection of antibiotics [57, 59].

3.4.2 Immuno-complex-based electrochemical biosensor

According to Wilson and Hu [60], electrochemical immunosensors that rely on particular antigen–antibody recognition have the advantage of being highly dependable, reasonably priced, and able to detect targets with great sensitivity, such as drugs, environmental contaminants, and veterinary residues in food. To electrochemically utilize extremely specific “antigen–antibody” interactions, however, electron mediators must be included in the detection process [61].

3.4.3 Aptamer-based electrochemical biosensor

Recently, electrochemical biosensors based on aptamers have become reliable methods for detecting antibiotic residues. Aptamers have superior physicochemical stability, an extended shelf life, facile in vitro production, and straightforward modification protocols in contrast to antibodies [62]. Two primary methods are usually used to immobilize aptasensors: (i) modifying a bio-functionalized sensor surface directly with aptamers utilizing appropriate linkers and (ii) modifying functionally active surfaces non-covalently with aptamers. For the electrochemical identification of antibiotics, nanomaterials like gold NPs and carbon fibers are commonly used to increase the sensitivity [63].

3.4.4 Electrochemical biosensor based on molecularly imprinted polymers (MIPs)

Molecular imprinted sensors are gaining popularity due to their ability to detect analyte molecules with unique receptors. MIPs are manufactured using sacrificial spacer procedures. Lian et al. describe that cross-linkers and functional monomers are polymerized with template molecules by covalent and non-covalent techniques [64]. Nanomaterials, such as magnetic nanomaterials, graphene oxide, Prussian blue catalytic polymer, and metal nanoparticles, have been widely used to improve the sensitivity of MIP-based electrochemical sensors for antibiotic residues [65, 66].

3.5 Different types of antibiotic detection by electrochemical methods

Nanomaterials are thought to be the most appealing alternative for developing and building “functional” transducers for electrochemical biosensing systems due to their distinct physical and chemical characteristics. The ability to modify or functionalize these systems in a variety of ways has significantly improved analytical performance for antibiotic identification. In light of this advancement, the next section will cover several nanomaterials used for electrochemical antibiotic detection (Table 1).

Order Sensing interfaceTransduction methodElectrodeTarget antibioticsMatricesLimit of detection (nM)Linear range (M)Sensitivity (μAμM 1 cm 2)References
ssDNA/SWCNTSWVAuLevofloxacinUrine75.21 × 10−6–10 × 10−6—[67]
ssDNA/AuNPs/en/MWCNTsCVAuValrubicinHuman urine & Blood samples180.5 × 10−6–80 × 10−6—[63]
Anti-TET/MWCNTsDPVGCETetracycline (TET)Spiked milk samples50.5 × 10−8–5 × 10−5—[68]
Au-PtNPs/MWCNTLSVGCECefotaximePlasma samples14 × 10−9–4 × 10−6, 4 × 10−6–10 × 10−64.76,1.3[69]
3DCNTs@Cu NPsCVGCEChloramphenicolMilk10,0001 × 10−5–5 × 10−4—[70]
SGN-hematein/ILs/penicillinaseDPVGCEPenicillinMilk samples0.00011.25 × 10−13–7.5 × 10−3—[71]

Table 1.

Evaluates the performance of several nanomaterials for electrochemical antibiotic sensing.

Carbon nanotubes are highly effective in biosensing applications. Carbon nanotubes have unique physical, chemical, and electrical characteristics, making them ideal for recognizing analytes, metabolites, and disease indicators [10]. Numerous studies have focused on nano-structuring with CNTs/CNT-composite electrodes used for antibiotic detection in electrochemical biosensors. Because nanomaterials enhanced electrochemical performance, common antibiotics were detected at lower detection limits and with a wider detection range [62, 63, 72].

Moraes et al. suggested using CNT-based electrochemical sensors to detect levofloxacin in human urine samples. The sensor detected 75.2 nM of levofloxacin in urine within a detection range of 1–10 μM. This suggests that MWCNTs oriented vertically should exhibit strong electrocatalytic activity in the electrooxidation of levofloxacin [67].

Another study used gold NPs and multiwalled CNTs (Au NPs/en/MWCNTs/Au) nanocomposites to create an electrochemical sensor for detecting valrubicin in both urine and blood samples [63]. Following the discovery of valrubicin in 1998, the work was described as a “stepping stone” toward its analytical identification. The study also reported varied DNA-valrubicin interactions via effective immobilization of several single-stranded DNA probe sequences on the surface of the composite. Consequently, sequences containing guanine and cytosine exhibited a greater affinity for valrubicin. With a detection range of 0.5–80.0 μM and a detection limit of 0.018 μM, the sensor demonstrated high valrubicin sensitivity.

Similarly, carboxyl functionalized multiwalled carbon nanotubes (MWCNT) were used to enhance the signal from an electrochemical aptasensor to detect tetracycline (TET). It has been demonstrated that MWCNTs add conductivity and biocompatibility, enabling a 5 nM TET detection limit (LOD) in milk samples that have been tampered with. Two chemically identical tetracycline derivatives (OTC and DOX) were present in real-world samples (such as milk) when the sensor’s response was assessed [68].

A very sensitive voltammetric cefotaxime detection device was developed by electrodepositing a mixture of multiwalled carbon nanotubes (MWCNTs)-based bimetallic Au-Pt nanoparticles [69]. Compared to GCE fabricated separately with Au-PtNPs or MWCNTs, this composite demonstrated a detection limit of 1.0 nM throughout a linear range (0.004–10 μM).

A 3-dimensional imprinted nanohybrid structure-based new sensing method was created employing 3DCNTs@Cu NPs nanocomposite. To detect chloramphenicol, a molecularly imprinted polymer (MIP)-based sensor with high sensitivity and a detection limit of 10 μM was constructed [70].

Graphene has gained significant attention in electronics and sensing. Research suggests that graphene nanosheets outperform single-walled carbon nanotubes in terms of conductivity, surface area, stability, and sensitivity [34, 73, 74].

Wu et al. presented a penicillin biosensor using layered films like nanosheets of graphene adsorbed with hematein, penicillinase, and ionic liquids. The sensor uses penicillinase enzyme to convert penicillin to penicilloic acid, and hematein as a pH indicator to monitor freed Hþ. This sensor has capability of detecting penicillin at ultralow concentrations (DL = 0.1 pM) [71].

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4. Conclusion

The development of electrochemical sensors for antibiotic detection has gained significant attention due to the rising concerns over antibiotic contamination and resistance. These sensors offer a promising solution for real-time, sensitive, and selective detection of antibiotics in various matrices, including environmental, clinical, and food samples. Advances in nanotechnology and material science, particularly the integration of nanomaterials like graphene, metal nanoparticles, and metal-organic frameworks, have further enhanced the performance of electrochemical sensors by improving their sensitivity, selectivity, and stability. Despite the remarkable progress, challenges remain, particularly in sensor durability, interference from complex sample matrices, and the need for more cost-effective and scalable production methods. Furthermore, future research should focus on developing portable and user-friendly devices that can be employed in field applications, ensuring broad accessibility and ease of use. In conclusion, electrochemical sensors are poised to play a critical role in monitoring and controlling antibiotic residues and resistance, contributing to both public health and environmental safety. Continued research and interdisciplinary collaboration will be key to overcoming current limitations and fully realizing the potential of these sensors in real-world applications.

References

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Rafia Masood, Abdur Rahim, Abdul Wajid, Sana Sabahat, Zia Ul Haq Khan, Abdul Qadeer and Hafiz Irshadullah

Submitted: 13 September 2024 Reviewed: 21 September 2024 Published: 19 December 2024

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