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New cosmetic formulations are continuously requested by the market and the ingredients are constantly evolving. Recently the use of antioxidants has gained success and, in this context, analytical methods able to quickly and easily assess the antioxidant activity of cosmetics would make it possible to carry out analyses on new formulations even within the manufacturing process without the need for specialized laboratories and personnel, thus evaluating directly on-site the effectiveness and the shelf life of products. In this work, a chemiluminescent inhibition assay was developed for determining the total antioxidant activity in cosmetic products. The method was based on the luminol/enhancers/hydrogen peroxide/horseradish peroxidase chemiluminescent system, which generates light signals measurable through simple and compact instrumentation. The formation of the chemiluminescent signal is inhibited by the presence of antioxidant substances while it is restored once all the antioxidant molecules have been oxidized. The time of appearance of the light signal is related to the total antioxidant activity. The assay was carried out exploiting an integrated device comprising a microwell plate coupled with an array of amorphous silicon hydrogenated photosensors enclosed in a mini-dark box. The method was optimized in terms of concentrations and volumes of the required reagents and sample pre-treatment. A calibration curve was generated taking as a reference the antioxidant activity of ascorbic acid obtaining a detection limit of 10 µM. The developed method was applied to cosmetic products currently on the market as well as on spiked samples in order to evaluate the performance of the methods in terms of sensitivity, accuracy, and reproducibility. [ABSTRACT FROM AUTHOR]

Analytical chemistry applied to medical and diagnostic analysis has recently focused on the development of cost-effective biosensors able to monitor the health status or to assess the level of specific biomarkers that can be indicative of several diseases. The improvement of technologies relating to the possibility of the non-invasive sampling of biological fluids, as well as sensors for the detection of analytical signals and the computational capabilities of the systems routinely employed in everyday life (e.g., smartphones, computers, etc.), makes the complete integration of self-standing analytical devices more accessible. This review aims to discuss the biosensors that have been proposed in the last five years focusing on two principal detecting approaches, optical and electrochemical, which have been employed for quantifying different kinds of target analytes reaching detection limits below the clinical sample levels required. These detection principles applied to point-of-care (POC) devices have been extensively reported in literature, and even the limited examples found on the market are based on these strategies. This work will show the latest innovations considering the integration of optical and electrochemical detection with the most commonly reported analytical platforms for POC applications such as paper-based or wearable and implantable devices. [ABSTRACT FROM AUTHOR]

Water is indispensable for life, yet many lack access to clean drinking water, resulting in fatalities from waterborne bacterial infections. Precise assessment of microbial abundance and viability in natural aquatic environments is vital. Adenosine triphosphate (ATP) serves as a parameter for viability assessments due to its presence in viable bacterial cells as an energy carrier. Traditional ATP detection methods involve chemical or enzymatic extraction, followed by measurement of light emission via the Luciferin–Luciferase complex. However, these methods are costly, present a low stability, require specialized equipment, and entail complex sample pretreatment. To overcome these limitations, we developed a biosensor based on aptamers, nucleic acid sequences with specific target-molecule-binding capabilities. Aptamers offer advantages such as an enhanced stability, a lower cost, and ease of design compared to antibodies. Recently, ATP has been used for aptamer selection testing. Our proposed biosensor utilizes a structure-switching ATP-binding DNA nanoswitch with two functional domains: a catalytic DNA-zyme domain and an ATP-binding aptamer domain. In the presence of ATP, its binding to the aptamer domain triggers the activation of the DNA-zyme domain, which is exploited for chemiluminescence (CL) detection. Integrating functional DNA biosensors with microfluidic paper-based analytical devices (µPADs) holds promise for point-of-care (POC) applications. However, achieving proper DNA binding on paper remains challenging, often requiring solution-based assay protocols, leaving µPADs for final signal readout. Here, we introduce an origami µPAD with preloaded dried reagents, allowing for on-paper assay execution upon sample addition and proper folding. Paper functionalization strategies and assay protocols were optimized to ensure simple and straightforward detection of ATP, employing a portable charge-coupled device (CCD) camera for CL detection. Calibration curves plotted against the logarithm of ATP concentration in the range of 1 to 500 µM facilitated determination of the assay's limit of detection (LOD), which was found to be 3 µM. [ABSTRACT FROM AUTHOR]

The impurities present in recombinant protein drugs produced by large-scale refolding processes can not only affect the product safety but also interact with the expressed protein. To relate the impurity profile to conformation and functionality of the protein drug, analytical methods able not to degrade the sample components should be preferred. In this work, an urate oxidase (uricase) drug from Aspergillus flavus expressed in Saccharomyces cerevisiae, and a reagent-grade uricase from Candida sphaerica expressed in Escherichia coli, are analyzed by combining hollow-fiber flow field-flow fractionation with matrix-assisted laser desorption-ionization rime-of-flight mass spectrometry (MALDI/TOFMS) and with chemiluminescence enzyme activity assay. Preliminary detection and identification of sample impurities is performed by means of conventional methods such as RP HPLC with electrospray ionization quadrupole-TOF MS and MALDI/TOFMS with SDS PAGE and 2D SDS PAGE. Results show that the recombinant uricase samples obtained from different microorganisms have different impurities and different enzymatic activity and that different uricase oligomers are present in solution.