Electrochemical Biosensors for the Detection of Antibiotics in Milk: Recent Trends and Future Perspectives.

Antibiotics have emerged as ground-breaking medications for the treatment of infectious diseases, but due to the excessive use of antibiotics, some drugs have developed resistance to microorganisms. Because of their structural complexity, most antibiotics are excreted unchanged, polluting the water, soil, and natural resources. Additionally, food items are being polluted through the widespread use of antibiotics in animal feed. The normal concentrations of antibiotics in environmental samples typically vary from ng to g/L. Antibiotic residues in excess of these values can pose major risks the development of illnesses and infections/diseases. According to estimates, 300 million people will die prematurely in the next three decades (by 2050), and the WHO has proclaimed "antibiotic resistance" to be a severe economic and sociological hazard to public health. Several antibiotics have been recognised as possible environmental pollutants (EMA) and their detection in various matrices such as food, milk, and environmental samples is being investigated. Currently, chromatographic techniques coupled with different detectors (e.g., HPLC, LC-MS) are typically used for antibiotic analysis. Other screening methods include optical methods, ELISA, electrophoresis, biosensors, etc. To minimise the problems associated with antibiotics (i.e., the development of AMR) and the currently available analytical methods, electrochemical platforms have been investigated, and can provide a cost-effective, rapid and portable alternative. Despite the significant progress in this field, further developments are necessary to advance electrochemical sensors, e.g., through the use of multi-functional nanomaterials and advanced (bio)materials to ensure efficient detection, sensitivity, portability, and reliability. This review summarises the use of electrochemical biosensors for the detection of antibiotics in milk/milk products and presents a brief introduction to antibiotics and AMR followed by developments in the field of electrochemical biosensors based on (i) immunosensor, (ii) aptamer (iii) MIP, (iv) enzyme, (v) whole-cell and (vi) direct electrochemical approaches. The role of nanomaterials and sensor fabrication is discussed wherever necessary. Finally, the review discusses the challenges encountered and future perspectives. This review can serve as an insightful source of information, enhancing the awareness of the role of electrochemical biosensors in providing information for the preservation of the health of the public, of animals, and of our environment, globally.

Potentiometric sensing of histamine using immobilized enzymes on layered double hydroxides.

Diamine oxydase and peroxidase have been co-immobilized onto layered double hydroxide (LDH) thin films for the development of real-time histamine biosensors. The chosen LDH materials are Mg2AlCO3, Mg4FeCl and Ca2AlCl. Prepared bi-enzymatic hybrid nanomaterials are capable of detecting histamine through the electrochemical oxidation of H2O2 and are used as the sensitive membrane for potentiometric microelectrode. Histamine biosensors developed in this work have fast response of less than 20 s, are sensitive and selective, with a large dynamic range of 10-8-10-3 M and a limit of detection of less than 10-8 M. The detection limit of the developed bi-enzymatic biosensors is relatively higher than those corresponding with gas and liquid chromatography, which are still considered as the reference methods. Finally, the reproducibility, the specificity and the storage stability of the biosensors were studied.

Sensitivity and specificity improvement of an ion sensitive field effect transistors-based biosensor for potato glycoalkaloids detection.

Butyryl cholinesterase of different origin along with variations of the time of enzyme immobilization on the potentiometric transducer surface is offered to control the ion sensitive field effect transistor (ISFET)-based biosensor sensitivity. Because butyryl cholinesterase has been already used to develop the sensors for heavy metals, organophosphorus/carbamate pesticides, and steroidal glycoalkaloids analysis, the present study has been focused on the investigation and adjustment of the ISFET-based biosensor specificity exclusively to the glycoalkaloids. Utilization of ethylendiaminetetracetate (a complexon of heavy metal ions) and phosphotriesterase (a highly efficient catalyst for the hydrolysis of organophosphorus compounds) enabled the highly specific determination of glycoalkaloids at the background of lead and mercury (up to 500 microM of ions concentration) and paraoxon (up to 100 microM of pesticide concentration). The developed biosensor has been validated for glycoalkaloids detection in potato varieties cultivated in Ukraine, and the results obtained are compared to those measured by the methods of HPLC and TLC.

Potato glycoalkaloids: true safety or false sense of security?

As one of the major agricultural crops, the cultivated potato is consumed each day by millions of people from diverse cultural backgrounds. A product of global importance, the potato tuber contains toxic glycoalkaloids (GAs) that cause sporadic outbreaks of poisoning in humans, as well as many livestock deaths. This article will discuss some aspects of the potato GAs, including their toxic effects and risk factors, methods of detection of GAs and biotechnological aspects of potato breeding. An attempt has been made to answer a question of vital importance - are potato GAs dangerous to humans and animals and, if so, to what extent?

Biosensors based on enzyme field-effect transistors for determination of some substrates and inhibitors.

This paper is a review of the authors' publications concerning the development of biosensors based on enzyme field-effect transistors (ENFETs) for direct substrates or inhibitors analysis. Such biosensors were designed by using immobilised enzymes and ion-selective field-effect transistors (ISFETs). Highly specific, sensitive, simple, fast and cheap determination of different substances renders them as promising tools in medicine, biotechnology, environmental control, agriculture and the food industry. The biosensors based on ENFETs and direct enzyme analysis for determination of concentrations of different substrates (glucose, urea, penicillin, formaldehyde, creatinine, etc.) have been developed and their laboratory prototypes were fabricated. Improvement of the analytical characteristics of such biosensors may be achieved by using a differential mode of measurement, working solutions with different buffer concentrations and specific agents, negatively or positively charged additional membranes, or genetically modified enzymes. These approaches allow one to decrease the effect of the buffer capacity influence on the sensor response in an aim to increase the sensitivity of the biosensors and to extend their dynamic ranges. Biosensors for the determination of concentrations of different toxic substances (organophosphorous pesticides, heavy metal ions, hypochlorite, glycoalkaloids, etc.) were designed on the basis of reversible and/or irreversible enzyme inhibition effect(s). The conception of an enzymatic multibiosensor for the determination of different toxic substances based on the enzyme inhibition effect is also described. We will discuss the respective advantages and disadvantages of biosensors based on the ENFETs developed and also demonstrate their practical application.

Metabolically engineered methylotrophic yeast cells and enzymes as sensor biorecognition elements.

An extended definition of the term metabolic engineering is given and its successful use in the construction of biorecognition elements of sensors is demonstrated. It is shown that genetic and chemical modifications of methylotrophic yeast cells provide directed changes in their physiological responses towards methanol, ethanol and formaldehyde resulting in enhanced selectivity and shorter time response of the corresponding potentiometric and amperometric biosensors.