Xylenes are ubiquitous in the environment due to their widespread use in a variety of products and industries, including cleaning agents, paint thinners, printing, and plastic production, demonstrating the potential for exposure in humans. Because of limited data on xylenes, we are investigating the toxicity of xylenes, mixed in a specific isomeric ratio, in rodent models. The goal of this study was to develop and validate a method to quantitate individual xylene isomers (p-, m-, and o-xylene) in rodent blood in support of toxicity studies. The method used internal standard calibration with isotopically labeled analytes for an analysis by headspace gas chromatography-mass spectrometry following removal of moisture with anhydrous calcium chloride to allow separation of individual isomers. In male Sprague Dawley rat blood, the method was linear ( ≥ 0.99) over the range of 5 - 1500 ng/mL, accurate (mean relative error (RE) ≤ 15.3%), and precise (relative standard deviation (RSD) ≤ 10.8%). Low limits of detection were achieved, at 1.0, 2.3, and 1.0 ng/mL for p-, m-, and o-xylene, respectively. The method was evaluated in male and female Hsd:Sprague Dawley SD rat blood and male and female B6C3F1/N mouse blood, the strains used in toxicology studies, with RE ≤ ±13.5% and RSD ≤ 10.6%. All isomers were stable for at least 62 days in blood when stored frozen. The results presented here demonstrate this method can accurately and precisely quantitate p-, m-, and o-xylene in a single analytical run in rat and mice blood samples following exposure to individual xylene isomers. The method could be easily adapted to other biological matrices, including human matrices.

Ondansetron is an anti-emetic drug and has been recently identified as a therapeutic for neuropathic pain. Permeability of ondansetron into the nervous system may be impacted by the presence of P-glycoprotein (Pgp) at the blood brain barrier, since ondansetron is a substrate of Pgp, which means it may not easily cross into the brain. In order to determine a drug concentration within the brain, microdialysis can be used, which measures unbound analyte concentrations in tissues or body fluids.

While many chemicals are regulated and routinely monitored in drinking water, they represent just a portion of all contaminants that may be present. Typical drinking water analyses involve sampling one liter or less of water, which could lead to trace level contaminants going undetected. In this study, a method was developed for using point-of-use activated carbon block drinking water filters as sampling devices. The filters were extracted to remove chemicals that were collected, and then analyzed by non-targeted analysis via liquid chromatography and gas chromatography high-resolution mass spectrometry. Extraction efficiencies were assessed by spiking and recovery experiments to better understand the chemical space coverage. To test the method's applicability to real-world samples, filters from a small-scale pilot study were collected from individuals in New York, NY and Atlanta, GA and analyzed. Twenty tentatively identified chemical candidates were confirmed by comparison to chemical standards. Principal components analysis was performed on the full set of filtered chemical features to explore how geographic and temporal differences in samples impact drinking water composition. Product use categories for confirmed chemicals were explored to determine potential sources of contaminants.

, a versatile human pathogen, significantly impacts global health causing a broad spectrum of medical conditions that range from minor skin infections to life-threatening diseases. The clinical importance of is underscored by its resistance to multiple antibiotics and formation of biofilms, providing protection against antimicrobials and immune responses. To date, the identification of antimicrobial-resistant (AMR) strains, such as methicillin-resistant (MRSA) and vancomycin-intermediate (VISA), requires time-consuming and expensive methodologies, including culture-based, molecular, and phenotypic techniques. Previously, we developed a paper-based ratiometric sensor array composed of fluorescent sensor dyes (3-hydroxyflavone derivatives) pre-adsorbed on paper microzone plates. Combined with machine learning algorithms such as neural networks, this sensor effectively discriminated 16 bacterial species and determined their Gram status. In this study, we evaluate its ability to distinguish antibiotic-resistant strains and their biofilms. Our results demonstrate that the sensor array, in conjunction with LDA and neural networks, successfully differentiated three common laboratory MRSA strains from three methicillin-susceptible (MSSA) strains with 82.5% accuracy. Furthermore, using support vector machines, this sensor was able to distinguish and categorically classify MRSA, MSSA, and VISA clinical isolates with 97.5% accuracy. Remarkably, beyond distinguishing planktonic cultures, this sensor array demonstrated a formidable capability to discriminate AMR biofilms, achieving over 80% accuracy. Combined, the results of this study highlight the paper-based sensor array's significant potential as a robust diagnostic tool to accurately, rapidly, and easily identify drug-resistant strains in clinically relevant settings.

Tibetan strawberry () is a wild medicinal and edible plant in Tibet possessing various health benefits such as neuroprotection and anti-oxidation. However, there has been little study reported on its chemical constituents. To investigate the inhibitors of monoamine oxidase B (MAO-B) in Tibetan strawberry, we immobilized the enzyme onto cellulose filter paper for the first time to develop a new screening method. Two known glycosides (compounds and ) and one new iridoid glucoside (Compound ) were fished out by this method, which was found to effectively inhibit MAO-B with IC values of 16.95 ± 0.93, 24.69 ± 0.20, and 46.77 ± 0.78 μM, respectively. Molecular docking and kinetic analysis were performed to reveal the inhibition mechanism of these compounds. Furthermore, compound exhibited neuroprotective effects against 6-OHDA-induced injury on PC12 cells. The developed method exhibits the advantages of rapidness and effectiveness in screening of MAO-B inhibitors from complex herbal extracts.

A sensitive chemiluminescent enzyme immunoassay (CLEIA) was established for the determination of gentamicin (GEN) residue levels in animal tissue. This assay is based on a fusion protein of single-chain variable fragment (scFv) and alkaline phosphatase (AP). Initially, V and V derived from anti-gentamicin monoclonal antibody were linked by a short peptide to construct a scFv. Subsequently, the constructed scFv sequence was accessed into the pLIP6/GN vector, and a soluble scFv-AP fusion protein was generated. The scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) for the determination of gentamicin. In the dcCLEIA, the half inhibitory concentration (IC) and limit of detection (LOD) were 1.073 ng/mL and 0.380 ng/mL, respectively. The average recoveries of gentamicin spiked in animal tissue samples ranged from 78% to 96%. These results showed a strong correlation with ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The above results suggest that the anti-GEN scFv-AP fusion protein is suitable for detecting gentamicin residues in edible animal tissues.

The SARS-CoV-2 pandemic has resulted in a devastating effect on human health in the last three years. While tremendous effort has been devoted to the development of effective treatment and vaccines against SARS-CoV-2 and controlling the spread of it, collective health challenges have been encountered along with the concurrent serious economic impacts. Since the beginning of the pandemic, various detection methods like PCR-based methods, isothermal nucleic acid amplification-based (INAA) methods, serological methods or antibody tests, and evaluation of X-ray chest results have been exploited to diagnose SARS-CoV-2. PCR-based detection methods in these are considered gold standards in the current stage despite their drawbacks, including being high-cost and time-consuming procedures. Furthermore, the results obtained from the PCR tests are susceptible to sample collection methods and time. When the sample is not collected properly, obtaining a false result may be likely. The use of specialized lab equipment and the need for trained people for the experiments pose additional challenges in PCR-based testing methods. Also, similar problems are observed in other molecular and serological methods. Therefore, biosensor technologies are becoming advantageous with their quick response, high specificity and precision, and low-cost characteristics for SARS-CoV-2 detection. In this paper, we critically review the advances in the development of sensors for the detection of SARS-CoV-2 using two-dimensional (2D) materials. Since 2D materials including graphene and graphene-related materials, transition metal carbides, carbonitrides, and nitrides (MXenes), and transition metal dichalcogenides (TMDs) play key roles in the development of novel and high-performance electrochemical (bio)sensors, this review pushes the sensor technologies against SARS-CoV-2 detection forward and highlights the current trends. First, the basics of SARS-CoV-2 detection are described. Then the structure and the physicochemical properties of the 2D materials are explained, which is followed by the development of SARS-CoV-2 sensors by exploiting the exceptional properties of the 2D materials. This critical review covers most of the published papers in detail from the beginning of the outbreak.

A portable, field deployable whole-cell biosensor was developed that can withstand the complex matrices of soil and requires minimal to no sample preparation to monitor bioavailable concentrations of the essential micronutrient copper (II). Conventional measurement of micronutrients is often complex, laboratory-based, and not suitable for monitoring their bioavailable concentration. To address this need, we developed a fluorescence based microbial whole-cell biosensing (MWCB) system encoding for a Cu-responsive protein capable of generating a signal upon binding to Cu. The sensing-reporting protein was designed by performing circular permutation on the green fluorescent protein (GFP) followed by insertion of a Cu binding motif into the structure of GFP. The design included insertion of several binding motifs and creating plasmids that encoded the corresponding sensing proteins. The signal generated by the sensing-reporting protein is directly proportional to the concentration of Cu in the sample. Evaluation of the resulting biosensing systems carrying these plasmids was performed prior to selection of the optimal fluorescence emitting Cu-binding protein. The resulting optimized biosensing system was encapsulated in polyacrylate-alginate beads and embedded in soil for detection of the analyte. Once exposed to the soil, the beads were interrogated to measure the fluorescence signal emitted by the sensing-reporting protein using a portable imaging device. The biosensor was optimized for detection of Cu in terms of selectivity, sensitivity, matrix effects, detection limits, and reproducibility in both liquid and soil matrices. The limit of detection (LoD) of the optimized encapsulated biosensor was calculated as 0.27 mg/L and 1.26 mg/kg of Cu for Cu in solution and soil, respectively. Validation of the portable imaging tools as a potential biosensing device in the field was performed.

Traditional quantum dot-based lateral flow immunoassay (QD-LFIA) is limited to signal loss in part by the blinking, photobleaching and oxidative quenching of QD probes. Inspired by the good application of silver deposition on QD surfaces in tissue imaging, and in the context of improving the assay performance without compromising the simplicity and practicality, we report that introducing the QD-silver combination to the LFIA system, has the advantages of accuracy improvement, signal enhancement and user friendliness promotion, but maintains the cost-effective property and commercial accessibility of QD-LFIA. The effect was shown by using CdSe/ZnS QD-LFIA coupled with anti-sodium pentachlorophenate antibody, which provided a 4-fold improvement in the signal, a 2.5-fold improvement in the detection limit and a zero false-negative rate for sodium pentachlorophenate analysis in chicken samples. The proposed LFIA integrates the possibilities of colorimetric and fluorometric detection with different detection limits (fluorometric at 10 ng/mL and colorimetric at 4 ng/mL) and with acceptable detection times (fluorometric at 12 min and colorimetric at 27 min). The current results indicate that this QD-silver combined LFIA is complementary to conventional fluorescence LFIA and could be an inexpensive, versatile, and sensitive alternative.

Favipiravir (FVP) is introduced as a promising newly developed antiviral drug against the coronavirus disease 2019 (COVID-19). Therefore, the accurate determination of FVP is of great significance for quality assessment and clinical diagnosis. Herein, a novel electrochemical sensing platform for FVP based on gold nanoparticles anchored conductive carbon black (Au@CCB) modified graphite nanopowder flakes paste electrode (GNFPE) was constructed. Morphological and nanostructure properties of Au@CCB have been investigated by TEM, HRTEM, and EDX methods. The morphology and electrochemical properties of Au@CCB/GNFPE were characterized by SEM, cyclic voltammetry (CV), and EIS. The Au@CCB nanostructured modified GNFPE exhibited strong electro-catalytic ability towards the oxidation of FVP. The performance of the fabricated Au@CCB/GNFPE was examined by monitoring FVP concentrations in the absence and presence of co-administered drug paracetamol (PCT) by AdS-SWV. It was demonstrated that the proposed sensor exhibited superior sensitivity, stability, and anti-interference capability for the detection of FVP. The simultaneous determination of a binary mixture containing FVP and the co-administered drug PCT using Au@CCB/GNFPE sensor is reported for the first time. Under optimized conditions, the developed sensor exhibited sensitive voltammetric responses to FVP and PCT with low detection limits of 7.5 nM and 4.3 nM, respectively. The sensing electrode was successfully used to determine FVP and PCT simultaneously in spiked human plasma and pharmaceutical preparations, and the findings were satisfactory. Finally, the fabricated sensor exhibited high sensitivity for simultaneous detection of FVP and PCT in the presence of ascorbic acid in a real sample.

Establishing sensitive and targeted analytical methodologies for drug identification in biological fluids as well as screening of treatments that can counteract the most severe COVID-19 infection-related side effects are of utmost importance. Here, first attempts have been made for determination of the anti-COVID drug Remdesivir (RDS) in human plasma using four potentiometric sensors. Calixarene-8 (CX8) was used as an ionophore applied to the first electrode (Sensor I). The second had a layer of dispersed graphene nanocomposite coating (Sensor II). (Sensor III) was fabricated using nanoparticles of polyaniline (PANI) as ion-to-electron transducer. A reverse-phase polymerization using polyvinylpyrrolidone (PVP) was employed to create a graphene-polyaniline (G/PANI) nanocomposite electrode (Sensor IV). Surface morphology was confirmed by Scanning Electron Microscope (SEM). UV absorption spectra and Fourier Transform Ion Spectrophotometry (FTIR) also supported their structural characterization. The impact of graphene and polyaniline integration on the functionality and durability of the manufactured sensors was examined using the water layer test and signal drift. In the ranges of concentration of 10 to 10 mol/L and 10 to 10, sensors II & IV exhibited linear responses; respectively while sensors I & III displayed linearity within 10 to 10 mol/L. The target drug was easily detectable using LOD down to 100 nmol/L. The developed sensors satisfactorily offered sensitive, stable, selective and accurate estimate of Remdesivir (RDS) in its pharmaceutical formulation as well as spiked human plasma with recoveries ranging from 91.02 to 95.76 % with average standard deviations less than 1.85. The suggested procedure was approved in accordance with ICH recommendations.

While monitoring expression of recombinant proteins is essential for obtaining high-quality biopharmaceutical and biotechnological products, existing assays for recombinant protein detection are laborious, time-consuming and expensive. This paper presents a microfluidic approach to rapid and cost-effective detection of tag-fused recombinant proteins via a dual-aptamer sandwich assay. Our approach addresses limitations in current methods for both dual-aptamer assays and generation of aptamers for such assays by first using microfluidic technology to isolate the aptamers rapidly and then employing these aptamers to implement a microfluidic dual-aptamer assay for tag-fused recombinant protein detection. The use of microfluidic technology enables the fast generation of aptamers and rapid detection of recombinant proteins with minimized consumption of reagents. In addition, compared with antibodies, aptamers as low-cost affinity reagents with an ability of reversible denaturation further decreases the cost of recombinant protein detection. For demonstration, an aptamer pair is isolated rapidly toward His-tagged IgE within two days, and then used in the microfluidic dual-aptamer assay for detecting His-tagged IgE in cell culture media within 10 min and with a limit of detection of 7.1 nM.

A novel fluorometric strategy for the simultaneous identification of SARS-CoV-2 and SARS-CoV was successfully established based on a hybridization-induced signal on-off-on mechanism. Here, one part of the probe (P1) of SARS-CoV-2 (P = P1/P2) is partially related to SARS-CoV, while the other part (P2) is completely irrelevant to SARS-CoV. They as smart gatekeepers were anchored on NH-MIL-88(Fe) (MOF@P1/P2) to turn off its catalytic performance. Only the specific SARS-CoV-2 genetic target can strongly restore the peroxidase-like activity of MOF@P1/P2. In the presence of o-phenylenediamine, SARS-CoV-2 can be efficiently detected with high sensitivity, accuracy, and reliability. This strategy demonstrated excellent analytical characteristics with a linear range (10 M ∼ 10 M) under the limit of detection of 0.11 nM not only in buffer but also in 10 % serum, which partly shows its practicability. Most importantly, with the help of the auxiliary test of MOF@P1 and MOF@P2, SARS-CoV-2 and SARS-CoV can be efficiently quantified and distinguished. This novel strategy has provided a breakthrough in the development of such identification. In the whole process, only a simple one-step experiment was involved. This circumvents the trouble of pretreatment experiments in traditional methods, including complex enzymatic mixtures, specialized experimental equipment, many primers optimization as well as reverse transcriptase. Additionally, this novel strategy is rapid, low-cost, and easy-to-use tools.

Platinum nanoparticles (PtNPs) have been attracted worldwide attention due to their versatile application potentials, especially in the catalyst and sensing fields. Herein, a facile synthetic method of triethanolamine (TEOA)-capped PtNPs (TEOA@PtNP) for electrochemiluminescent (ECL) and colorimetric immunoassay of SARS-CoV spike proteins (SARS-CoV S-protein, a target detection model) is developed. Monodisperse PtNPs with an average diameter of 2.2 nm are prepared by a one-step hydrothermal synthesis method using TEOA as a green reductant and stabilizer. TEOA@PtNPs can be used as a nanocarrier to combine with antigen by the high-affinity antibody, which leads to a remarkable inhibition of electron transfer efficiency and mass transfer processes. On the basis of its peroxidase-like activity and easy-biolabeling property, the TEOA@PtNP can be used to establish a colorimetric immunosensor of SARS-CoV S-protein thought catalyzing the reaction of HO and 3,3',5,5'-tetramethylbenzidine (TMB). Especially, the Ru(bpy) ECL reaction is well-achieved with the TEOA@PtNPs due to their great conductivity and loading abundant TEOA co-reactants, resulting in an enhancing ECL signal in immunoassay of SARS-CoV S-protein. As a consequence, two proposed methods could achieve sensitive detection of SARS-CoV S-protein in wide ranges, the colorimetric and ECL detection limits were as low as 8.9 fg /mL and 4.2 fg /mL (S/N = 3), respectively. We believe that the proposed colorimetric and ECL immunosesors with high sensitivity, good reproducibility, and good stability will be a promising candidate for a broad spectrum of applications.

Coronavirus disease 2019 (COVID-19) and influenza A are two respiratory infectious diseases with similar clinical manifestations. Because of the complex global epidemic situation of COVID-19, the distinction and diagnosis of COVID-19 and influenza A infected persons is crucial for epidemic prevention and control. In this study, tetrahedral DNA framework (TDF) was combined with a rotational paper-based analytical device, and the color change generated by the reaction between horseradish peroxidase (HRP) and 3,3'5,5'-tetramethylbenzidine (TMB)-HO was used for grayscale signal analysis by ImageJ software. The quantitative detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A H1N1 virus were realized simultaneously. Under the optimal conditions, the paper-based analytical device showed a good linear relationship between the two viruses in the range of 10-10g/mL, and the two viruses were not affected by cross reaction. This sensor provides a convenient and reliable method for clinical rapid differentiation and diagnosis of COVID-19 and influenza A.

Hydroxychloroquine is a drug that has been widely used during the early stages of COVID-19 pandemic. Different liquid chromatographic methods have been reported for the analysis of hydroxychloroquine in various biological matrices such as human plasma, serum, whole blood, oral fluid, rat plasma and tissues. In this comparative study, the most popular tools used for assessing the greenness profile: National Environmental Methods Index (NEMI), Eco-Scale Assessment (ESA), Green Analytical Procedure Index (GAPI) and Analytical Greenness metric (AGREE) were utilized to evaluate the ecological impact of eighteen liquid chromatographic methods developed for the bioanalysis of COVID-19 drug; hydroxychloroquine. NEMI is the simplest tool for evaluating the greenness profile of developed methods, but it is the least informative approach as all the reported methods had the same NEMI pictograms. On the other hand, GAPI is a dependable tool providing a complete picture about the method greenness starting from sampling until the final determination. ESA and AGREE tools are digitally presented and more easily applied. Therefore, their utilization for greenness assessment is highly recommended. Selection of the highest eco-friendly analytical procedure is of a paramount importance for protecting human health and the environment. Considering the greenness of the analytical procedures is highly recommended before proceeding to routine use in order to minimize the chemical hazards to the environment. The most eco-friendly analytical procedures for the analysis of hydroxychloroquine in biological samples according to ESA, GAPI and AGREE tools will be highlighted and discussed.

To reduce the progression of the viral process in patients infected with COVID-19, new treatments and drug active substances are needed. One of these drugs is Molnupiravir (MNP) which has a direct antiviral effect and has also proven to be highly effective in reducing the azopharyngeal SARS-CoV-2 infectious virus and viral RNA. Due to the importance and frequent use of this drug in the treatment of COVID-19, its accurate, quick, and cheap detection in pharmaceutical or biological samples is crucial. In this work, electrochemical behavior and sensitive voltammetric determination of MNP are described using a magnetite nanoparticle modified carbon paste electrode (FeO@CPE) for the first time. FeO nanoparticles (NPs) were characterized by recording their transmission electron microscopy (TEM) images, energy dispersive X-ray (EDX), and X-ray diffraction (XRD) spectra. Cyclic voltammetric measurements showed that MNP was irreversibly oxidized at FeO@CPE at 760 mV in pH 2.0 Britton Robinson buffer solution (BRBS). The peak current of MNP was increased approximately threefold at FeO@CPE compared to bare CPE due to a good electrocatalytic efficiency of FeO NPs. According to differential pulse voltammetric studies, the fabricated electrode exhibited a linear range (LR) between 0.25 and 750 µM with sensitivity and limit of detection (LOD) of 4591.0 µA mM cm and 0.05 µM, respectively. On the other hand, although lower sensitivity (327.3 µA mM cm) was obtained from CV compared to DPV, a wider linear calibration curve between 0.25 and 1500 µM was obtained in CV. Studies performed in tablet samples confirmed that the FeO@CPE exhibits high applicability for selective and accurate voltammetric determination of MNP in real samples.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-COV-2) is a cluster of β coronaviruses. The 2019 coronavirus disease (COVID-19) caused by SARS-COV-2 is emerging as a global pandemic. Thus, early diagnosis of SARS-COV-2 is essential to prevent severe outbreaks of the disease. In this experiment, a novel label-free photoelectrochemical (PEC) immunosensor was obtained based on silver sulfide (AgS) sensitized titanium dioxide@bismuth tungstate (TiO@BiWO) nanocomposite for quantitative detection of SARS-COV-2 nucleocapsid protein. The constructed TiO@BiWO hollow microspheres had large specific surface area and could produce high photocurrent intensity under visible light illumination. AgS was in-situ grown on the surface of thioglycolic acid (TGA) modified TiO@BiWO. In particular, TiO@BiWO and AgS formed a good energy level match, which could effectively enhance the photocurrent conversion efficiency and strength the photocurrent response. Ascorbic acid (AA) acted as an effective electron donor to effectively eliminate photogenerated holes. Under optimal experimental conditions, the constructed immunosensor presented a supersensitive response to SARS-COV-2 nucleocapsid protein, with a desirable linear relationship ranged from 0.001 to 50 ng/mL for nucleocapsid protein and a lower detection limit of 0.38 pg/mL. The fabricated sensor exhibited a wide linear range, excellent selectivity, specificity and stability, which provided a valuable referential idea for the detection of SARS-COV-2.

A sensitive and selective UPLC-MS/MS method was developed for the synchronized determination of four drugs used in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), namely, azithromycin, apixaban, dexamethasone, and favipiravir in rat plasma. using a Poroshell 120 EC-C18 column (50 mm × 4.6 mm, 2.7 m) with a high-resolution ESI tandem mass spectrometer detection with multiple reaction monitoring. We used an Agilent Poroshell column, which is characterized by a stationary phase based on non-porous core particles. With a remarkable improvement in the number of theoretical plates and low column backpressure. In addition, the developed method was employed in studying the potential food-drug interaction of grapefruit juice (GFJ) with the selected drugs which affects their pharmacokinetics in rats. The LC-MS/MS operated in positive and negative ionization mode using two internal standards: moxifloxacin and chlorthalidone, respectively. Liquid- liquid extraction of the cited drugs from rat plasma was accomplished using diethyl ether: dichloromethane (70:30, ). The analytes were separated using methanol: 0.1 % formic acid in water (95: 5, ) as a mobile phase in isocratic mode of elution pumped at a flow rate of 0.3 mL/min. A detailed validation of the bio-analytical method was performed in accordance with US-FDA and EMA guidelines. Concerning the pharmacokinetic study, the statistical significance between the results of the test groups receiving GFJ along with the cited drugs and the control group was assessed demonstrating that GFJ increased the plasma concentration of azithromycin, apixaban, and dexamethasone. Accordingly, this food-drug interaction requires cautious ingestion of GFJ in patients using (SARS-CoV-2) medications as it can produce negative effects in the safety of the drug therapy. A potential drug-drug interaction is also suggested between those medications requiring a suitable dose adjustment.

This work describes the development of a Point-of-Care (POC) Lab-on-a-Chip (LOC) instrument for diagnosis of SARS-CoV-2 by Reverse-Transcription Loop-mediated isothermal amplification (RT-LAMP). The hardware is based on a Raspberry Pi computer ($35), a video camera, an Arduino Nano microcontroller, a printed circuit board as a heater and a 3D printed housing. The chips were manufactured in polymethyl methacrylate (PMMA) using a CO2 laser cutting machine and sealed with a PCR optic plastic film. The chip temperature is precisely controlled by a proportional-integral-derivative (PID) algorithm. During the RT-LAMP amplifications the chip was maintained at ∼ (65.0 ± 0.1) °C for 25 minutes and 5 minutes cooling down, totaling a 30 minutes of reaction .The software interpretation occurs in less than a second. The chip design has four 25 µL chambers, two for clinical samples and two for positive and negative control-samples. The RT-LAMP master mix solution added in the chip chambers contains the pH indicator Phenol Red, that is pink (for pH ∼ 8.0) before amplification and becomes yellow (pH ∼ 6.0) if the genetic material is amplified. The RT-LAMP SARS-CoV-2 diagnostic was made by color image recognition using the OpenCV machine vision software library. The software was programmed to automatically distinguish the HSV color parameter distribution in each one of the four chip chambers. The instrument was successfully tested for SARS-CoV-2 diagnosis, in 22 clinic samples, 11 positives and 11 negatives, achieving an assertiveness of 86% when compared to the results obtained by RT-LAMP standard reactions performed in conventional PCR equipment.

Innovative therapeutic protocols to the rapidly spreading coronavirus disease (COVID19) epidemic is highly required all across the world. As demonstrated by clinical studies, Favipiravir (FVP) and Remdesivir (REM) are new antiviral medicines that are effective against COVID-19. REM is the first FDA approved antiviral medicine against COVID-19. In addition to antivirals, corticosteroids such as dexamethasone (DEX), and anticoagulants such as apixaban (PX) are used in multidrug combinations protocols. This work develops and validates simple and selective screening of the four medicines of COVID -19 therapeutic protocol. FVP, REM, DEX, and PX as internal standard in human plasma using UPLC method by C18 column and methanol, acetonitrile, and water acidified by orthophosphate (pH = 4) in a ratio of (15: 35: 50, by volume) as an eluate flowing at 0.3 mL/min. The eluent was detected at 240 nm. The method was linear over (0.1-10 μg/mL) for each of FVP, REM, and DEX. The validation of the UPLC method was assessed in accordance with FDA guidelines. The method can detect as low as down to 0.1 μg/mL for all. The recoveries of the drugs in spiked human plasma ranged from 97.67 to 102.98 percent. Method accuracy and precision were assessed and the drugs showed good stability. The method was proven to be green to the environment after greenness checking by greenness profile and Eco-Scale tool.