Search results for: analytes

Authors: Sze Shin Low, Michelle T. T. Tan, Poi Sim Khiew, Hwei-San Loh

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An efficient graphene/zinc oxide (PSE-G/ZnO) platform based on pi-pi stacking, non-covalent interactions for the development of aptamer-based biosensor was presented in this study. As a proof of concept, the DNA recognition capability of the as-developed PSE-G/ZnO enhanced aptamer-based biosensor was evaluated using Coconut Cadang-cadang viroid disease (CCCVd). The G/ZnO nanocomposite was synthesised via a simple, green and efficient approach. The pristine graphene was produced through a single step exfoliation of graphite in sonochemical alcohol-water treatment while the zinc nitrate hexahydrate was mixed with the graphene and subjected to low temperature hydrothermal growth. The developed facile, environmental friendly method provided safer synthesis procedure by eliminating the need of harsh reducing chemicals and high temperature. The as-prepared nanocomposite was characterised by X-ray diffractometry (XRD), scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) to evaluate its crystallinity, morphology and purity. Electrochemical impedance spectroscopy (EIS) was employed for the detection of CCCVd sequence with the use of potassium ferricyanide (K3[Fe(CN)6]). Recognition of the RNA analytes was achieved via the significant increase in resistivity for the double stranded DNA, as compared to single-stranded DNA. The PSE-G/ZnO enhanced aptamer-based biosensor exhibited higher sensitivity than the bare biosensor, attributing to the synergistic effect of high electrical conductivity of graphene and good electroactive property of ZnO.

Keywords: aptamer-based biosensor, graphene/zinc oxide nanocomposite, green synthesis, screen printed carbon electrode

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Authors: Shalini Singh, Sanjay K. Srivastava, Govind, Mukhtar. A. Khan, P. K. Singh

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Nanotechnology is revolutionizing the development of biosensors. Nanomaterials and nanofabrication technologies are increasingly being used to design novel biosensors. Sensitivity and other attributes of biosensors can be improved by using nanomaterials with unique chemical, physical, and mechanical properties in their construction. Silicon is a promising biomaterial that is non-toxic and biodegradable and can be exploited in chemical and biological sensing. Present study demonstrated the streptavidin–biotin interaction on silicon surfaces with different topographies such as flat and nanostructured silicon (nanowires) surfaces. Silicon nanowires with wide range of surface to volume ratio were prepared by electrochemical etching of silicon wafer. The large specific surface of silicon nanowires can be chemically modified to link different molecular probes (DNA strands, enzymes, proteins and so on), which recognize the target analytes, in order to enhance the selectivity and specificity of the sensor device. The interaction of streptavidin with biotin was carried out on 3-aminopropyltriethoxysilane (APTS) functionalized silicon surfaces. Fourier Transform Infrared Spectroscopy (FTIR) and X-ray Photoelectron Spectroscopy (XPS) studies have been performed to characterize the surface characteristics to ensure the protein attachment. Silicon nanowires showed the enhance protein attachment, as compared to flat silicon surface due to its large surface area and good molecular penetration to its surface. The methodology developed herein could be generalized to a wide range of protein-ligand interactions, since it is relatively easy to conjugate biotin with diverse biomolecules such as antibodies, enzymes, peptides, and nucleotides.

Keywords: FTIR, silicon nanowires, streptavidin-biotin, XPS

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Authors: Diana A. Dobreva, Veselina Panayotova, Albena Merdzhanova, Lubomir Makedonski, Mona Stancheva

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Many studies suggest that marine mollusks are healthy food, characterized by low fat and high digestible proteins content. They are one of the most important dietary sources of fat soluble vitamins. The most common species of mollusks in the Bulgarian Black Sea waters are the black mussel (Mytilus galloprovincialis) and the sea snail Rapana (Rapana venosa). One of the main problems of the region is the lack of information about chemical composition of these important marine species. Due to these facts, the aim of the present work was to determine the fat soluble vitamins A, D2, D3, and E, carotenoids–β-carotene and astaxanthin, and total cholesterol contents of mollusk samples and compare them to sample of green algae (Ulva rigida). Samples were collected during autumn from north region of the Black Sea coast, and their wet tissues were used for evaluation of vitamins A, D2, D3, and E, astaxanthin, β-carotene and cholesterol compositions. All fat soluble analytes were simultaneously analyzed by RP- HPLC/UV/FL system. The results were calculated as milligrams per gram total lipid (mg.g-1TL). Alpha-tocopherol and b-carotene were most abundant in algae samples, while mussel samples presented the highest amounts of vitamin D3 (several times higher than the recommended daily intake in Bulgaria (Ordinance № 23 / 19.07.2005)). In all samples, cholesterol content was significantly low, which falls within recommendation of the same ordinance (upper daily consumption should not exceed 300 mg per day). From data, it can be concluded that all samples were characterized as beneficial sources of biologically active compounds.

Keywords: fat soluble vitamins, carotenoids, mussel, rapana, algae

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Authors: Andile Mkhohlakali, Tien-Chien Jen, James Tshilongo, Happy Mabowa

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Among different parameters, oxygen is one of the most important analytes of interest, dissolved oxygen (DO) concentration is very crucial and significant for various areas of physical, chemical, and environmental monitoring. Herein we report oxygen-sensitive luminophores -based lanthanum(III) trifluoromethanesulfonate), [La]³⁺ was encapsulated into SiO₂-based xerogel matrix. The nanosensor is composed of organically modified silica nanoparticles, doped with the luminescent oxygen–sensitive lanthanum(III) trifluoromethanesulfonate complex. The precursor materials used for sensing film were triethyl ethoxy silane (TEOS) and (3-Mercaptopropyltriethoxysilane) (MPTMS- TEOS) used for SiO2-baed matrices. Brunauer–Emmett–Teller (BET), and BJH indicate that the SiO₂ transformed from microporous to mesoporous upon the addition of La³⁺ luminophore with increased surface area (SBET). The typical amorphous SiO₂ based xerogels were revealed with X-Ray diffraction (XRD) and Selected Area Electron Diffraction (SAED) analysis. Scanning electron microscope- (SEM) and transmission electron microscope (TEM) showed the porous morphology and reduced particle for SiO₂ and La-SiO₂ xerogels respectively. The existence of elements, siloxane networks, and thermal stability of xerogel was confirmed by energy dispersive spectroscopy (EDS), Fourier-transform infrared spectroscopy (FTIR), and Thermographic analysis (TGA). UV-Vis spectroscopy and photoluminescence (PL) have been used to characterize the optical properties of xerogels. La-SiO₂ demonstrates promising characteristic features of an active sensing film for dissolved oxygen in the water. Keywords: Sol-gel, ORMOSILs, encapsulation, Luminophores quenching, O₂-sensing

Keywords: sol-gel, ORMOSILs, luminophores quenching, O₂-sensing

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Authors: F. Z. Tighilt, N. Belhaneche-Bensemra, S. Belhousse, S. Sam, K. Lasmi, N. Gabouze

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Recently, the gold nanoparticles (Au NPs) became an active multidisciplinary research topic. First, Au thin films fabricated by alkylthiol-functionalized Au NPs were found to have vapor sensitive conductivities, they were hence widely investigated as electrical chemiresistors for sensing different vapor analytes and even organic molecules in aqueous solutions. Second, Au thin films were demonstrated to have speciallocalized surface plasmon resonances (LSPR), so that highly ordered 2D Au superlattices showed strong collective LSPR bands due to the near-field coupling of adjacent nanoparticles and were employed to detect biomolecular binding. Particularly when alkylthiol ligands were replaced by thiol-terminated polymers, the resulting polymer-modified Au NPs could be readily assembled into 2D nanostructures on solid substrates. Monolayers of polystyrene-coated Au NPs showed typical dipolar near-field interparticle plasmon coupling of LSPR. Such polymer-modified Au nanoparticle films have an advantage that the polymer thickness can be feasibly controlled by changing the polymer molecular weight. In this article, the effect of tin-doped indium oxide (ITO) coatings on the plasmonic properties of ITO interfaces modified with gold nanostructures (Au NSs) is investigated. The interest in developing ITO overlayers is multiple. The presence of a con-ducting ITO overlayer creates a LSPR-active interface, which can serve simultaneously as a working electrode in an electro-chemical setup. The surface of ITO/ Au NPs contains hydroxyl groups that can be used to link functional groups to the interface. Here the covalent linking of nickel /Au NSs/ITO hybrid LSPR platforms will be presented.

Keywords: conducting polymer, metal nanoparticles (NPs), LSPR, poly (3-(pyrrolyl)–carboxylic acid), polypyrrole

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Authors: Vaibhav Budhiraja, Chandra Mouli Pandey

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The ultrasonic synthesis of nanostructured conducting copolymer is an effective technique to synthesize polymer with desired chemical properties. This tailored nanostructure, shows tremendous improvement in sensitivity and stability to detect a variety of analytes. The present work reports ultrasonically synthesized nanostructured conducting poly(o-phenylenediamine)-co-poly(1-naphthylamine) (POPD-co-PNA). The synthesized material has been characterized using Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy, transmission electron microscopy, X-ray diffraction and cyclic voltammetry. FTIR spectroscopy confirmed random copolymerization, while UV-visible studies reveal the variation in polaronic states upon copolymerization. High crystallinity was achieved via ultrasonic synthesis which was confirmed by X-ray diffraction, and the controlled morphology of the nanostructures was confirmed by transmission electron microscopy analysis. Cyclic voltammetry shows that POPD-co-PNA has rather high electrochemical activity. This behavior was explained on the basis of variable orientations adopted by the conducting polymer chains. The synthesized material was electrophoretically deposited at onto indium tin oxide coated glass substrate which is used as cathode and parallel platinum plate as the counter electrode. The fabricated bioelectrode was further used for detection of glucose by crosslinking of glucose oxidase in the PODP-co-PNA film. The bioelectrode shows a surface-controlled electrode reaction with the electron transfer coefficient (α) of 0.72, charge transfer rate constant (ks) of 21.77 s⁻¹ and diffusion coefficient 7.354 × 10⁻¹⁵ cm²s⁻¹.

Keywords: conducting, electrophoretic, glucose, poly (o-phenylenediamine), poly (1-naphthylamine), ultrasonic

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Authors: Gaurav Bhanjana, Ganga Ram Chaudhary, Sandeep Kumar, Neeraj Dilbaghi

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Main sources of endocrine disrupting compounds in the ecosystem are hormones, pesticides, phthalates, flame retardants, dioxins, personal-care products, coplanar polychlorinated biphenyls (PCBs), bisphenol A, and parabens. These endocrine disrupting compounds are responsible for learning disabilities, brain development problems, deformations of the body, cancer, reproductive abnormalities in females and decreased sperm count in human males. Although discharge of these chemical compounds into the environment cannot be stopped, yet their amount can be retarded through proper evaluation and detection techniques. The available techniques for determination of these endocrine disrupting compounds mainly include high performance liquid chromatography (HPLC), mass spectroscopy (MS) and gas chromatography-mass spectrometry (GC–MS). These techniques are accurate and reliable but have certain limitations like need of skilled personnel, time consuming, interference and requirement of pretreatment steps. Moreover, these techniques are laboratory bound and sample is required in large amount for analysis. In view of above facts, new methods for detection of endocrine disrupting compounds should be devised that promise high specificity, ultra sensitivity, cost effective, efficient and easy-to-operate procedure. Nowadays, electrochemical sensors/biosensors modified with nanomaterials are gaining high attention among researchers. Bioelement present in this system makes the developed sensors selective towards analyte of interest. Nanomaterials provide large surface area, high electron communication feature, enhanced catalytic activity and possibilities of chemical modifications. In most of the cases, nanomaterials also serve as an electron mediator or electrocatalyst for some analytes.

Keywords: electrochemical, endocrine disruptors, microscopy, nanoparticles, sensors

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Authors: Valeriy M. Kondratev, Ekaterina A. Vyacheslavova, Talgat Shugabaev, Alexander S. Gudovskikh, Alexey D. Bolshakov

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Modern sensorics imposes strict requirements on the biosensors characteristics, especially technological feasibility, and selectivity. There is a growing interest in the analysis of human health biological markers, which indirectly testifying the pathological processes in the body. Such markers are acids and alkalis produced by the human, in particular - ammonia and hydrochloric acid, which are found in human sweat, blood, and urine, as well as in gastric juice. Biosensors based on modern nanomaterials, especially low dimensional, can be used for this markers detection. Most classical adsorption sensors based on metal and silicon oxides are considered non-selective, because they identically change their electrical resistance (or impedance) under the action of adsorption of different target analytes. This work demonstrates a feasible frequency-resistive method of electrical impedance spectroscopy data analysis. The approach allows to obtain of selectivity in adsorption sensors of a resistive type. The method potential is demonstrated with analyzis of impedance spectra of silicon nanowires in the presence of NH3 and HCl vapors with concentrations of about 125 mmol/L (2 ppm) and water vapor. We demonstrate the possibility of unambiguous distinction of the sensory signal from NH3 and HCl adsorption. Moreover, the method is found applicable for analysis of the composition of ammonia and hydrochloric acid vapors mixture without water cross-sensitivity. Presented silicon sensor can be used to find diseases of the gastrointestinal tract by the qualitative and quantitative detection of ammonia and hydrochloric acid content in biological samples. The method of data analysis can be directly translated to other nanomaterials to analyze their applicability in the field of biosensory.

Keywords: electrical impedance spectroscopy, spectroscopy data analysis, selective adsorption sensor, nanotechnology

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Authors: Narayana Murthy Akurathi, Vijaya Lakshmi Marella

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The literature is reviewed with regard to online Super critical fluid extraction (SFE) coupled directly with supercritical fluid chromatography (SFC) -mass spectrometry that have typically more sensitive than conventional LC-MS/MS and GC-MS/MS. It is becoming increasingly interesting to use on-line techniques that combine sample preparation, separation and detection in one analytical set up. This provides less human intervention, uses small amount of sample and organic solvent and yields enhanced analyte enrichment in a shorter time. The sample extraction is performed under light shielding and anaerobic conditions, preventing the degradation of thermo labile analytes. It may be able to analyze compounds over a wide polarity range as SFC generally uses carbon dioxide which was collected as a by-product of other chemical reactions or is collected from the atmosphere as it contributes no new chemicals to the environment. The diffusion of solutes in supercritical fluids is about ten times greater than that in liquids and about three times less than in gases which results in a decrease in resistance to mass transfer in the column and allows for fast high resolution separations. The drawback of SFC when using carbon dioxide as mobile phase is that the direct introduction of water samples poses a series of problems, water must therefore be eliminated before it reaches the analytical column. Hundreds of compounds analysed simultaneously by simple enclosing in an extraction vessel. This is mainly applicable for pharmaceutical industry where it can analyse fatty acids and phospholipids that have many analogues as their UV spectrum is very similar, trace additives in polymers, cleaning validation can be conducted by putting swab sample in an extraction vessel, analysing hundreds of pesticides with good resolution.

Keywords: super critical fluid extraction (SFE), super critical fluid chromatography (SFC), LCMS/MS, GCMS/MS

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Authors: Zahrasadat Hosseini, Jie Yuan

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Point-of-care (POC) diagnostic devices based on lab-on-a-chip (LOC) technology have the potential to revolutionize medical diagnostics. However, the development of an ideal microfluidic system based on LOC technology for diagnostics purposes requires overcoming several obstacles, such as improving sensitivity, selectivity, portability, cost-effectiveness, and prototyping methods. While numerous studies have introduced technologies and systems that advance these criteria, existing systems still have limitations. Electrochemical enzyme-linked immunosorbent assay (e-ELISA) in a LOC device offers numerous advantages, including enhanced sensitivity, decreased turnaround time, minimized sample and analyte consumption, reduced cost, disposability, and suitability for miniaturization, integration, and multiplexing. In this study, we present a novel design and fabrication method for a microfluidic diagnostic platform that integrates screen-printed electrochemical carbon/silver chloride electrodes on flexible printed circuit boards with flexible, multilayer, polydimethylsiloxane (PDMS) microfluidic networks to accurately manipulate and pre-immobilize analytes for performing electrochemical enzyme-linked immunosorbent assay (e-ELISA) for multiplexed quantification of blood serum biomarkers. We further demonstrate fast, cost-effective prototyping, as well as accurate and reliable detection performance of this device for quantification of interleukin-6-spiked samples through electrochemical analytics methods. We anticipate that our invention represents a significant step towards the development of user-friendly, portable, medical-grade, POC diagnostic devices.

Keywords: lab-on-a-chip, point-of-care diagnostics, electrochemical ELISA, biomarker quantification, fast prototyping

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Authors: Zahrasadat Hosseini, Jie Yuan

Abstract:

Point-of-care (POC) diagnostic devices based on lab-on-a-chip (LOC) technology have the potential to revolutionize medical diagnostics. However, the development of an ideal microfluidic system based on LOC technology for diagnostics purposes requires overcoming several obstacles, such as improving sensitivity, selectivity, portability, cost-effectiveness, and prototyping methods. While numerous studies have introduced technologies and systems that advance these criteria, existing systems still have limitations. Electrochemical enzyme-linked immunosorbent assay (e-ELISA) in a LOC device offers numerous advantages, including enhanced sensitivity, decreased turnaround time, minimized sample and analyte consumption, reduced cost, disposability, and suitability for miniaturization, integration, and multiplexing. In this study, we present a novel design and fabrication method for a microfluidic diagnostic platform that integrates screen-printed electrochemical carbon/silver chloride electrodes on flexible printed circuit boards with flexible, multilayer, polydimethylsiloxane (PDMS) microfluidic networks to accurately manipulate and pre-immobilize analytes for performing electrochemical enzyme-linked immunosorbent assay (e-ELISA) for multiplexed quantification of blood serum biomarkers. We further demonstrate fast, cost-effective prototyping, as well as accurate and reliable detection performance of this device for quantification of interleukin-6-spiked samples through electrochemical analytics methods. We anticipate that our invention represents a significant step towards the development of user-friendly, portable, medical-grade POC diagnostic devices.

Keywords: lab-on-a-chip, point-of-care diagnostics, electrochemical ELISA, biomarker quantification, fast prototyping

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Authors: Yi-Shuan ShaoShao, Yen-An Tsai, Chia-Huang Chang, Kai-Wei Liao, Ming-Song Tsai, Mei-Lien Chen

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Organophosphate pesticides (OPs) is an environmental hormone with proven endocrine-disrupting effects that may affect the growth and development in human. A large amount of organophosphate pesticides (OPs) is used throughout Taiwan, and human may be exposed through dietary intake or residential use. During pregnancy, OPs can be transferred to the blood stream reaching the fetus through the placenta. The aim of this study was to explore the association between maternal OPs exposure levels and fetal developments and birth outcomes. A birth cohort was follow-up. Maternal urine sample were collected at the first, second, and third gestational trimester. Fetal growth characteristics were measured by ultrasonic scan and birth outcomes were assessed by pediatrician. Urinary metabolite of organophosphate pesticides were assessed using gas chromatography-mass spectrometry. The analytes included dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphates (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). We found that all of urine samples in each trimester were detected at least one metabolite for dialkyl phosphate (DAP). The detection rate range of OP urinary metabolites were from the lowest 22% DEDTP to the highest 100% DMP and DMTP. And to compared geometric means (GM) of urinary metabolites with three trimesters, that third trimester had the highest concentration for DMPs, DEPs, and DAPs in pregnant women were 368.01, 169.85 and 543.75 nmol/g creatinine, respectively. We observed that DAPs concentration in first and second trimester were significantly negative association with head circumference. DMPs in first trimester was significantly negative association with thoracic circumference (p=0.05) by spearman correlation. Our results support associations with prenatal OPs exposure with fetal head circumference and thoracic circumference. It provided that maternal OPs exposure might affect birth outcomes. Thus, prenatal exposure to OPs and health risk worthy of attention and concern.

Keywords: DAPs, birth outcomes, organophosphate pesticides, prenatal

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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Authors: Eiman A. Al-Enezi, Gin Jose, Sikha Saha, Paul Millner

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Rare earth nanotechnology has gained a considerable amount of interest in the field of biosensing due to the unique luminescence properties of lanthanides. Chelating rare earth ions plays a significant role in biological labelling applications including medical diagnostics, due to their different excitation and emission wavelengths, variety of their spectral properties, sharp emission peaks and long fluorescence lifetimes. We aimed to develop a platform for biosensors based on Europium (Eu³⁺) chelates against biomarkers of cardiac injury (heart-type fatty acid binding protein; H-FABP3) and stroke (glial fibrillary acidic protein; GFAP). Additional novelty in this project is the use of synthetic binding proteins (Affimers), which could offer an excellent alternative targeting strategy to the existing antibodies. Anti-GFAP and anti-HFABP3 Affimer binders were modified to increase the number of carboxy functionalities. Europium nitrate then incubated with the modified Affimer. The luminescence characteristics of the Eu³⁺ complex with modified Affimers and antibodies against anti-GFAP and anti-HFABP3 were measured against different concentrations of the respective analytes on excitation wavelength of 395nm. Bovine serum albumin (BSA) was used as a control against the IgG/Affimer Eu³⁺ complexes. The emission spectrum of Eu³⁺ complex resulted in 5 emission peaks ranging between 550-750 nm with the highest intensity peaks were at 592 and 698 nm. The fluorescence intensity of Eu³⁺ chelates with the modified Affimer or antibodies increased significantly by 4-7 folder compared to the emission spectrum of Eu³⁺ complex. The fluorescence intensity of the Affimer complex was quenched proportionally with increased analyte concentration, but this did not occur with antibody complex. In contrast, the fluorescence intensity for Eu³⁺ complex increased slightly against increased concentration of BSA. These data demonstrate that modified Affimers Eu³⁺ complexes can function as nanobiosensors with potential diagnostic and analytical applications.

Keywords: lanthanides, europium, chelates, biosensors

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Authors: Kathryn Rooney, Kaitlyn Eddy, Evan Landers, Weihui Li

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The international mortality rate for neonates and infants has been declining at a disproportionally low rate when compared to the overall decline in child mortality in recent decades. A significant portion of infant deaths could be prevented with the implementation of low-cost and easy to use physiological monitoring devices, by enabling early identification of symptoms before they progress into life-threatening illnesses. The oral diagnostic device discussed in this paper serves to continuously monitor the key vital signs of body temperature, respiratory rate, heart rate, and oxygen saturation. The device mimics an infant pacifier, designed to be easily tolerated by infants as well as orthodontically inert. The fundamental measurements are gathered via thermistors and a pulse oximeter, each encapsulated in medical-grade silicone and wired internally to a microcontroller chip. The chip then translates the raw measurements into physiological values via an internal algorithm, before outputting the data to a liquid crystal display screen and an Android application. Additionally, a biological sample collection chamber is incorporated into the internal portion of the device. The movement within the oral chamber created by sucking on the pacifier-like device pushes saliva through a small check valve in the distal end, where it is accumulated and stored. The collection chamber can be easily removed, making the sample readily available to be tested for various diseases and analytes. With the vital sign monitoring and sample collection offered by this device, abnormal fluctuations in physiological parameters can be identified and appropriate medical care can be sought. This device enables preventative diagnosis for infants who may otherwise have gone undiagnosed, due to the inaccessibility of healthcare that plagues vast numbers of underprivileged populations.

Keywords: neonate mortality, infant mortality, low-cost diagnostics, vital signs, saliva testing, preventative care

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Authors: Fariba Tadayon, Elmira Hassanlou, Hasan Bagheri, Mostafa Jafarian

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Heavy metals are released into water, plants, soil, and food by natural and human activities. Lead has toxic roles in the human body and may cause serious problems even in low concentrations, since it may have several adverse effects on human. Therefore, determination of lead in different samples is an important procedure in the studies of environmental pollution. In this work, an ultrasonic assisted-ionic liquid based-liquid-liquid microextraction (UA-IL-DLLME) procedure for the determination of lead in zucchini, pomegranate, and lettuce has been established and developed by using flame atomic absorption spectrometer (FAAS). For UA-IL-DLLME procedure, 10 mL of the sample solution containing Pb2+ was adjusted to pH=5 in a glass test tube with a conical bottom; then, 120 μL of 1-Hexyl-3-methylimidazolium hexafluoro phosphate (CMIM)(PF6) was rapidly injected into the sample solution with a microsyringe. After that, the resulting cloudy mixture was treated by ultrasonic for 5 min, then the separation of two phases was obtained by centrifugation for 5 min at 3000 rpm and IL-phase diluted with 1 cc ethanol, and the analytes were determined by FAAS. The effect of different experimental parameters in the extraction step including: ionic liquid volume, sonication time and pH was studied and optimized simultaneously by using Response Surface Methodology (RSM) employing a central composite design (CCD). The optimal conditions were determined to be an ionic liquid volume of 120 μL, sonication time of 5 min, and pH=5. The linear ranges of the calibration curve for the determination by FAAS of lead were 0.1-4 ppm with R2=0.992. Under optimized conditions, the limit of detection (LOD) for lead was 0.062 μg.mL-1, the enrichment factor (EF) was 93, and the relative standard deviation (RSD) for lead was calculated as 2.29%. The levels of lead for pomegranate, zucchini, and lettuce were calculated as 2.88 μg.g-1, 1.54 μg.g-1, 2.18 μg.g-1, respectively. Therefore, this method has been successfully applied for the analysis of the content of lead in different food samples by FAAS.

Keywords: Dispersive liquid-liquid microextraction, Central composite design, Food samples, Flame atomic absorption spectrometry.

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Authors: Jaeyoung Kim, Heeju Lee, Huijin Jung, Heesoo Pyo, Seungki Kim, Joonseok Lee

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Avian influenza viruses (AIV) are the primary cause of highly contagious respiratory diseases caused by type A influenza viruses of the Orthomyxoviridae family. AIV are categorized on the basis of types of surface glycoproteins such as hemagglutinin and neuraminidase. Certain H5 and H7 subtypes of AIV have evolved to the high pathogenic avian influenza (HPAI) virus, which has caused considerable economic loss to the poultry industry and led to severe public health crisis. Several commercial kits have been developed for on-site detection of AIV. However, the sensitivity of these methods is too low to detect low virus concentrations in clinical samples and opaque stool samples. Here, we introduced a background-free near-infrared (NIR)-to-NIR upconversion nanoparticle-based lateral flow immunoassay (NNLFA) platform to yield a sensor that detects AIV within 20 minutes. Ca²⁺ ion in the shell was used to enhance the NIR-to-NIR upconversion photoluminescence (PL) emission as a heterogeneous dopant without inducing significant changes in the morphology and size of the UCNPs. In a mixture of opaque stool samples and gold nanoparticles (GNPs), which are components of commercial AIV LFA, the background signal of the stool samples mask the absorption peak of GNPs. However, UCNPs dispersed in the stool samples still show strong emission centered at 800 nm when excited at 980 nm, which enables the NNLFA platform to detect 10-times lower viral load than a commercial GNP-based AIV LFA. The detection limit of NNLFA for low pathogenic avian influenza (LPAI) H5N2 and HPAI H5N6 viruses was 10² EID₅₀/mL and 10³.⁵ EID₅₀/mL, respectively. Moreover, when opaque brown-colored samples were used as the target analytes, strong NIR emission signal from the test line in NNLFA confirmed the presence of AIV, whereas commercial AIV LFA detected AIV with difficulty. Therefore, we propose that this rapid and background-free NNLFA platform has the potential of detecting AIV in the field, which could effectively prevent the spread of these viruses at an early stage.

Keywords: avian influenza viruses, lateral flow immunoassay on-site detection, upconversion nanoparticles

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Authors: Nidhi Gupta, Omita Nanda, Rakhi Grover, Kanchan Saxena

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Hybrid perovskite is new class of material which has gained much attention due to their different crystal structure and interesting optical and electrical properties. Easy fabrication, high absorption coefficient, and photoluminescence properties make them a strong candidate for various applications such as sensors, photovoltaics, photodetectors, etc. In perovskites, ions arrange themselves in a special type of crystal structure with chemical formula ABX3, where A is organic species like CH3NH3+, B is metal ion (e.g., Pb, Sn, etc.) and X is halide (Cl-, Br-, I-). In crystal structure, A is present at corner position, B at center of the crystal lattice and halide ions at the face centers. High stability and sensitivity of nanostructured perovskite make them suitable for chemical sensors. Researchers have studied sensing properties of perovskites for number of analytes such as 2,4,6-trinitrophenol, ethanol and other hazardous chemical compounds. Ammonia being highly toxic agent makes it a reason of concern for the environment. Thus the detection of ammonia is extremely important. Our present investigation deals with organic inorganic hybrid perovskite based ammonia sensor. Various methods like sol-gel, solid state synthesis, thermal vapor deposition etc can be used to synthesize Different hybrid perovskites. In the present work, a novel hybrid perovskite has been synthesized by a single step method. Ethylenediammnedihalide and lead halide were used as precursor. Formation of hybrid perovskite was confirmed by FT-IR and XRD. Morphological characterization of the synthesized material was performed using scanning electron microscopy (SEM). SEM analysis revealed the formation of one dimensional nanowire perovskite with mean diameter of 200 nm. Measurements for sensing properties of halide perovskite for ammonia vapor were carried out. Perovskite thin films showed a color change from yellow to orange on exposure of ammonia vapor. Electro-optical measurements show that sensor based on lead halide perovskite has high sensitivity towards ammonia with effective selectivity and reversibility. Sensor exhibited rapid response time of less than 20 seconds.

Keywords: hybrid perovskite, ammonia, sensor, nanostructure, thin film

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Authors: Vijaya Madhuri Devraj, Swarnalatha Guditi, Kiran Kumar Bokara, Gangadhar Taduri

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Cell-based strategies may open therapeutic approaches that promote tolerance through manipulation of macrophages to increase long-term transplant survival rates and minimize side effects of the current immune suppressive regimens. The aim of the present study was, therefore, to test and compare the therapeutic potential of MSC and Tregs on macrophage polarization to develop an alternate cell-based treatment option in kidney transplantation. In the current protocol, macrophages from kidney transplant recipients with graft dysfunction were co-cultured with MSCs and Treg cells with and without cell-cell contact on transwell plates, further to quantitatively assess macrophage polarization in response to MSC and Treg treatment over time, M1 and M2 cell surface markers were used. Additionally, multiple soluble analytes were analyzed in cell supernatant by using bead-based immunoassays. Furthermore, to confirm our findings, gene expression analysis was done. MSCs induced the formation of M2 macrophages more than Tregs when macrophages M0 were cultured in transwell without cell contact. From this, we deduced the mechanism that soluble factors present in the MSCs condition media are involved in skewing of macrophages towards type 2 macrophages; similarly, in co-culture with cell-cell contact, MSCs resulted in more M2 type macrophages than Tregs. And an important finding of this study is the combination of both MSC-Treg showed significantly effective and consistent results in both with and without cell contact setups. Hence, it is suggestive to prefer MSCs over Tregs for secretome-based therapy and a combination of both for either therapy for effective transplantation outcomes. Our findings underline a key role of Tregs and MSCs in promoting macrophage polarization towards anti-inflammatory type. The study has great importance in prolongation of allograft and patient survival without any rejection by cell-based therapy, which induce self-tolerance and controlling infection.

Keywords: graft rejection, graft tolerance, macrophage polarization, mesenchymal stem cells, regulatory T cells, transplant immunology

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Authors: Mohamed Korany, Azza Gazy, Essam Khamis, Marwa Adel, Miranda Fawzy

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Background: Two new, simple and specific methods; First, a Zero-crossing first-derivative technique and second, a chemometric-assisted spectrophotometric artificial neural network (ANN) were developed and validated in accordance with ICH guidelines. Both methods were used for the simultaneous estimation of the two closely related antioxidant nutraceuticals ; Coenzyme Q10 (Q) ; also known as Ubidecarenone or Ubiquinone-10, and Vitamin E (E); alpha-tocopherol acetate, in their pharmaceutical binary mixture. Results: For first method: By applying the first derivative, both Q and E were alternatively determined; each at the zero-crossing of the other. The D1 amplitudes of Q and E, at 285 nm and 235 nm respectively, were recorded and correlated to their concentrations. The calibration curve is linear over the concentration range of 10-60 and 5.6-70 μg mL-1 for Q and E, respectively. For second method: ANN (as a multivariate calibration method) was developed and applied for the simultaneous determination of both analytes. A training set (or a concentration set) of 90 different synthetic mixtures containing Q and E, in wide concentration ranges between 0-100 µg/mL and 0-556 µg/mL respectively, were prepared in ethanol. The absorption spectra of the training sets were recorded in the spectral region of 230–300 nm. A Gradient Descend Back Propagation ANN chemometric calibration was computed by relating the concentration sets (x-block) to their corresponding absorption data (y-block). Another set of 45 synthetic mixtures of the two drugs, in defined range, was used to validate the proposed network. Neither chemical separation, preparation stage nor mathematical graphical treatment were required. Conclusions: The proposed methods were successfully applied for the assay of Q and E in laboratory prepared mixtures and combined pharmaceutical tablet with excellent recoveries. The ANN method was superior over the derivative technique as the former determined both drugs in the non-linear experimental conditions. It also offers rapidity, high accuracy, effort and money saving. Moreover, no need for an analyst for its application. Although the ANN technique needed a large training set, it is the method of choice in the routine analysis of Q and E tablet. No interference was observed from common pharmaceutical additives. The results of the two methods were compared together

Keywords: coenzyme Q10, vitamin E, chemometry, quantitative analysis, first derivative spectrophotometry, artificial neural network

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Authors: Monika Ksiezopolska-Gocalska, Pawel Albrycht, Robert Holyst

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Surface Enhanced Raman Spectroscopy (SERS) is a technique used to analyze very low concentrations of substances in solutions, even in aqueous solutions - which is its advantage over IR. This technique can be used in the pharmacy (to check the purity of products); forensics (whether at a crime scene there were any illegal substances); or medicine (serving as a medical test) and lots more. Due to the high potential of this technique, its increasing popularity in analytical laboratories, and simultaneously - the absence of appropriate platforms enhancing the SERS signal (crucial to observe the Raman effect at low analyte concentration in solutions (1 ppm)), we decided to invent our own SERS platforms. As an enhancing layer, we have chosen gold and silver nanoparticles, because these two have the best SERS properties, and each has an affinity for the other kind of particles, which increases the range of research capabilities. The next step was to commercialize them, which resulted in the creation of the company ‘SERSitive.eu’ focusing on production of highly sensitive (Ef = 10⁵ – 10⁶), homogeneous and reproducible (70 - 80%) substrates. SERStive SERS substrates are made using the electrodeposition of silver or silver-gold nanoparticles technique. Thanks to a very detailed analysis of data based on studies optimizing such parameters as deposition time, temperature of the reaction solution, applied potential, used reducer, or reagent concentrations using a standardized compound - p-mercaptobenzoic acid (PMBA) at a concentration of 10⁻⁶ M, we have developed a high-performance process for depositing precious metal nanoparticles on the surface of ITO glass. In order to check a quality of the SERSitive platforms, we examined the wide range of the chemical compounds and the biological substances. Apart from analytes that have great affinity to the metal surfaces (e.g. PMBA) we obtained very good results for those fitting less the SERS measurements. Successfully we received intensive, and what’s more important - very repetitive spectra for; amino acids (phenyloalanine, 10⁻³ M), drugs (amphetamine, 10⁻⁴ M), designer drugs (cathinone derivatives, 10⁻³ M), medicines and ending with bacteria (Listeria, Salmonella, Escherichia coli) and fungi.

Keywords: nanoparticles, Raman spectroscopy, SERS, SERS applications, SERS substrates, SERSitive

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Authors: Bharti Jain, Rajeev Jain, Abuzar Kabir, Torki Zughaibi, Shweta Sharma

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Drug-facilitated crimes (DFCs) entail the use of a single drug or a mixture of drugs to render a victim unable. Traditionally, biological samples have been gathered from victims and conducted analysis to establish evidence of drug administration. Nevertheless, the rapid metabolism of various drugs and delays in analysis can impede the identification of such substances. For this, the present article describes a rapid, sustainable, highly efficient and miniaturized protocol for the identification and quantification of three sedative-hypnotic drugs, namely diazepam, chlordiazepoxide and ketamine in alcoholic beverages and complex food samples (cream of biscuit, flavored milk, juice, cake, tea, sweets and chocolate). The methodology involves utilizing fabric phase sorptive extraction (FPSE) to extract diazepam (DZ), chlordiazepoxide (CDP), and ketamine (KET). Subsequently, the extracted samples are subjected to analysis using gas chromatography-mass spectrometry (GC-MS). Several parameters, including the type of membrane, pH, agitation time and speed, ionic strength, sample volume, elution volume and time, and type of elution solvent, were screened and thoroughly optimized. Sol-gel Carbowax 20M (CW-20M) has demonstrated the most effective extraction efficiency for the target analytes among all evaluated membranes. Under optimal conditions, the method displayed linearity within the range of 0.3–10 µg mL–¹ (or µg g–¹), exhibiting a coefficient of determination (R2) ranging from 0.996–0.999. The limits of detection (LODs) and limits of quantification (LOQs) for liquid samples range between 0.020-0.069 µg mL-¹ and 0.066-0.22 µg mL-¹, respectively. Correspondingly, the LODs for solid samples ranged from 0.056-0.090 µg g-¹, while the LOQs ranged from 0.18-0.29 µg g-¹. Notably, the method showcased better precision, with repeatability and reproducibility both below 5% and 10%, respectively. Furthermore, the FPSE-GC-MS method proved effective in determining diazepam (DZ) in forensic food samples connected to drug-facilitated crimes (DFCs). Additionally, the proposed method underwent evaluation for its whiteness using the RGB12 algorithm.

Keywords: drug facilitated crime, fabric phase sorptive extraction, food forensics, white analytical chemistry

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Authors: Pawel Albrycht, Monika Ksiezopolska-Gocalska, Robert Holyst

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Surface Enhanced Raman Spectroscopy (SERS) is a technique of growing importance not only in purely scientific research related to analytical chemistry. It finds more and more applications in broadly understood testing - medical, forensic, pharmaceutical, food - and everywhere works perfectly, on one condition that SERS substrates used for testing give adequate enhancement, repeatability, and homogeneity of SERS signal. This is a problem that has existed since the invention of this technique. Some laboratories use as SERS amplifiers colloids with silver or gold nanoparticles, others form rough silver or gold surfaces, but results are generally either weak or unrepeatable. Furthermore, these structures are very often highly specific - they amplify the signal only of a small group of compounds. It means that they work with some kinds of analytes but only with those which were used at a developer’s laboratory. When it comes to research on different compounds, completely new SERS 'substrates' are required. That underlay our decision to develop universal substrates for the SERS spectroscopy. Generally, each compound has different affinity for both silver and gold, which have the best SERS properties, and that's what depends on what signal we get in the SERS spectrum. Our task was to create the platform that gives a characteristic 'fingerprint' of the largest number of compounds with very high repeatability - even at the expense of the intensity of the enhancement factor (EF) (possibility to repeat research results is of the uttermost importance). As specified above SERS substrates are offered by SERSitive company. Applied method is based on cyclic potentiodynamic electrodeposition of silver or silver-gold nanoparticles on the conductive surface of ITO-coated glass at controlled temperature of the reaction solution. Silver nanoparticles are supplied in the form of silver nitrate (AgNO₃, 10 mM), gold nanoparticles are derived from tetrachloroauric acid (10 mM) while sodium sulfite (Na₂O₃, 5 mM) is used as a reductor. To limit and standardize the size of the SERS surface on which nanoparticles are deposited, photolithography is used. We secure the desired ITO-coated glass surface, and then etch the unprotected ITO layer which prevents nanoparticles from settling at these sites. On the prepared surface, we carry out the process described above, obtaining SERS surface with nanoparticles of sizes 50-400 nm. The SERSitive platforms present highly sensitivity (EF = 10⁵-10⁶), homogeneity and repeatability (70-80%).

Keywords: electrodeposition, nanoparticles, Raman spectroscopy, SERS, SERSitive, SERS platforms, SERS substrates

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Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

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A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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Authors: Meng-Ying Chiu, Yu-Fang Huang, Pei-Wei Wang, Yi-Ru Wang, Yi-Shuan Shao, Mei-Lien Chen

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Background: Organophosphate pesticides (OPs) are the most heavily used pesticides in agriculture in Taiwan. Therefore, they are commonly detected in general public including pregnant women and children. These compounds are proven endocrine disrupters that may affect the neural development in humans. The aim of this study is to assess the OPs exposure of children in 2 years of age and to examine the association between the exposure concentrations and neurodevelopmental effects in children. Methods: In a prospective cohort of 280 mother-child pairs, urine samples of prenatal and postnatal were collected from each participant and analyzed for metabolites of OPs by using gas chromatography-mass spectrometry. Six analytes were measured including dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). This study created a combined concentration measure for dimethyl compounds (DMs) consisting of the three dimethyl metabolites (DMP, DMTP, and DMDTP), for diethyl compounds (DEs) consisting of the three diethyl metabolites (DEP, DETP, and DEDTP) and six dialkyl phosphate (DAPs). The Bayley Scales of Infant and Toddler Development (Bayley-III) was used to assess children's cognitive behavior at 2 years old. The association between OPs exposure and Bayley-III scale score was determined by using the Mann-Whitney U test. Results: The measurements of urine samples are still on-going. This preliminary data are the report of 56 children aged 2 from the cohort. The detection rates for DMP, DMTP, DMDTP, DEP, DETP, and DEDTP are 80.4%, 69.6%, 64.3%, 64.3%, 62.5%, and 75%, respectively. After adjusting the creatinine concentrations of urine, the median (nmol/g creatinine) of urinary DMP, DMTP, DMDTP, DEP, DETP, DEDTP, DMs, DEs, and DAPs are 153.14, 53.32, 52.13, 19.24, 141.65, 192.17, 308.8, 311.6, and 702.11, respectively. The concentrations of urine are considerably higher than that in other countries. Children’s cognitive behavior was used three scales for Bayley-III, including cognitive, language and motor. In Mann-Whitney U test, the higher levels of DEs had significantly lower motor score (p=0.037), but no significant association was found between the OPs exposure levels and the score of either cognitive or language. Conclusion: The limited sample size suggests that Taipei children are commonly exposed to OPs and OPs exposure might affect the cognitive behavior of young children. This report will present more data to verify the results. The predictors of OPs concentrations, such as dietary pattern will also be included.

Keywords: biomonitoring, children, neurodevelopment, organophosphate pesticides exposure

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Authors: Adam Gushgari

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The SARS-CoV-2 pandemic has catalyzed the rapid adoption of wastewater-based epidemiology (WBE) methodologies both domestically and internationally. To support the rapid scale-up of pandemic-response wastewater surveillance systems, multiple federal agencies (i.e. US CDC), non-government organizations (i.e. Water Environment Federation), and private charities (i.e. Bill and Melinda Gates Foundation) have funded over $220 million USD supporting development and expanding equitable access of surveillance methods. Funds were primarily distributed directly to municipalities under the CARES Act (90.6%), followed by academic projects (7.6%), and initiatives developed by private companies (1.8%). In addition to federal funding for wastewater monitoring primarily conducted at wastewater treatment plants, state/local governments and private companies have leveraged wastewater sampling to obtain health and lifestyle data on student, prison inmate, and employee populations. We explore the viable paths for expansion of the WBE m1ethodology across a variety of analytical methods; the development of WBE-specific samplers and real-time wastewater sensors; and their application to various governments and private sector industries. Considerable investment in, and public acceptance of WBE suggests the methodology will be applied to other future notifiable diseases and health risks. Early research suggests that WBE methods can be applied to a host of additional “biological insults” including communicable diseases and pathogens, such as influenza, Cryptosporidium, Giardia, mycotoxin exposure, hepatitis, dengue, West Nile, Zika, and yellow fever. Interest in chemical insults is also likely, providing community health and lifestyle data on narcotics consumption, use of pharmaceutical and personal care products (PPCP), PFAS and hazardous chemical exposure, and microplastic exposure. Successful application of WBE to monitor analytes correlated with carcinogen exposure, community stress prevalence, and dietary indicators has also been shown. Additionally, technology developments of in situ wastewater sensors, WBE-specific wastewater samplers, and integration of artificial intelligence will drastically change the landscape of WBE through the development of “smart sewer” networks. The rapid expansion of the WBE field is creating significant business opportunities for professionals across the scientific, engineering, and technology industries ultimately focused on community health improvement.

Keywords: wastewater surveillance, wastewater-based epidemiology, smart cities, public health, pandemic management, substance abuse

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Authors: Irina Veselova, Maria Makedonskaya, Olga Eremina, Alexandr Sidorov, Eugene Goodilin, Tatyana Shekhovtsova

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Such catecholamines as dopamine, norepinephrine, and epinephrine are the principal neurotransmitters in the sympathetic nervous system. Catecholamines and their metabolites are considered to be important markers of socially significant diseases such as atherosclerosis, diabetes, coronary heart disease, carcinogenesis, Alzheimer's and Parkinson's diseases. Currently, neurotransmitters can be studied via electrochemical and chromatographic techniques that allow their characterizing and quantification, although these techniques can only provide crude spatial information. Besides, the difficulty of catecholamine determination in biological materials is associated with their low normal concentrations (~ 1 nM) in biomaterials, which may become even one more order lower because of some disorders. In addition, in blood they are rapidly oxidized by monoaminooxidases from thrombocytes and, for this reason, the determination of neurotransmitter metabolism indicators in an organism should be very rapid (15—30 min), especially in critical states. Unfortunately, modern instrumental analysis does not offer a complex solution of this problem: despite its high sensitivity and selectivity, HPLC-MS cannot provide sufficiently rapid analysis, while enzymatic biosensors and immunoassays for the determination of the considered analytes lack sufficient sensitivity and reproducibility. Fluorescent and SERS-sensors remain a compelling technology for approaching the general problem of selective neurotransmitter detection. In recent years, a number of catecholamine sensors have been reported including RNA aptamers, fluorescent ribonucleopeptide (RNP) complexes, and boronic acid based synthetic receptors and the sensor operated in a turn-off mode. In this work we present the fluorescent and SERS turn-on sensor systems based on the bio- or chemorecognizing nanostructured films {chitosan/collagen-Tb/Eu/Cu-nanoparticles-indicator reagents} that provide the selective recognition, visualization, and sensing of the above mentioned catecholamines on the level of nanomolar concentrations in biomaterials (cell cultures, tissue etc.). We have (1) developed optically transparent porous films and gels of chitosan/collagen; (2) ensured functionalization of the surface by molecules-'recognizers' (by impregnation and immobilization of components of the indicator systems: biorecognizing and auxiliary reagents); (3) performed computer simulation for theoretical prediction and interpretation of some properties of the developed materials and obtained analytical signals in biomaterials. We are grateful for the financial support of this research from Russian Foundation for Basic Research (grants no. 15-03-05064 a, and 15-29-01330 ofi_m).

Keywords: biomaterials, fluorescent and SERS-recognition, neurotransmitters, solid-phase turn-on sensor system

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Authors: Feiyu Yang, Chunfang Ni, Rong Wang, Yun Zou, Wenbin Liu, Chenggong Zhang, Fenjin Sun, Chun Wang

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Cocaine is the most frequently used illegal drug globally, with the global annual prevalence of cocaine used ranging from 0.3% to 0.4 % of the adult population aged 15–64 years. Growing consumption trend of abused cocaine and drug crimes are a great concern, therefore urine sample testing has become an important noninvasive sampling whereas cocaine and its metabolites (COCs) are usually present in high concentrations and relatively long detection windows. However, direct analysis of urine samples is not feasible because urine complex medium often causes low sensitivity and selectivity of the determination. On the other hand, presence of low doses of analytes in urine makes an extraction and pretreatment step important before determination. Especially, in gathered taking drug cases, the pretreatment step becomes more tedious and time-consuming. So developing a sensitive, rapid and high-throughput method for detection of COCs in human body is indispensable for law enforcement officers, treatment specialists and health officials. In this work, a new automated magnetic dispersive solid-phase extraction (MDSPE) sampling method followed by high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed for quantitative enrichment of COCs from human urine, using prepared magnetic nanoparticles as absorbants. The nanoparticles were prepared by silanizing magnetic Fe3O4 nanoparticles and modifying them with divinyl benzene and vinyl pyrrolidone, which possesses the ability for specific adsorption of COCs. And this kind of magnetic particle facilitated the pretreatment steps by electromagnetically controlled extraction to achieve full automation. The proposed device significantly improved the sampling preparation efficiency with 32 samples in one batch within 40mins. Optimization of the preparation procedure for the magnetic nanoparticles was explored and the performances of magnetic nanoparticles were characterized by scanning electron microscopy, vibrating sample magnetometer and infrared spectra measurements. Several analytical experimental parameters were studied, including amount of particles, adsorption time, elution solvent, extraction and desorption kinetics, and the verification of the proposed method was accomplished. The limits of detection for the cocaine and cocaine metabolites were 0.09-1.1 ng·mL-1 with recoveries ranging from 75.1 to 105.7%. Compared to traditional sampling method, this method is time-saving and environmentally friendly. It was confirmed that the proposed automated method was a kind of highly effective way for the trace cocaine and cocaine metabolites analyses in human urine.

Keywords: automatic magnetic dispersive solid-phase extraction, cocaine detection, magnetic nanoparticles, urine sample testing

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Authors: Mindaugas Liaudanskas, Darius Kviklys, Kristina Zymonė, Raimondas Raudonis, Jonas Viškelis, Norbertas Uselis, Pranas Viškelis, Valdimaras Janulis

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Apples are among the most commonly consumed fruits in the world. Based on data from the year 2014, approximately 84.63 million tons of apples are grown per annum. Apples are widely used in food industry to produce various products and drinks (juice, wine, and cider); they are also used unprocessed. Apples in human diet are an important source of different groups of biological active compounds that can positively contribute to the prevention of various diseases. They are a source of various biologically active substances – especially vitamins, organic acids, micro- and macro-elements, pectins, and phenolic, triterpenic, and other compounds. Triterpenic compounds, which are characterized by versatile biological activity, are the biologically active compounds found in apples that are among the most promising and most significant for human health. A specific analytical procedure including sample preparation and High Performance Liquid Chromatography (HPLC) analysis was developed, optimized, and validated for the detection of triterpenic compounds in the samples of different apples, their peels, and flesh from widespread apple cultivars 'Aldas', 'Auksis', 'Connel Red', 'Ligol', 'Lodel', and 'Rajka' grown in Lithuanian climatic conditions. The conditions for triterpenic compound extraction were optimized: the solvent of the extraction was 100% (v/v) acetone, and the extraction was performed in an ultrasound bath for 10 min. Isocratic elution (the eluents ratio being 88% (solvent A) and 12% (solvent B)) for a rapid separation of triterpenic compounds was performed. The validation of the methodology was performed on the basis of the ICH recommendations. The following characteristics of validation were evaluated: the selectivity of the method (specificity), precision, the detection and quantitation limits of the analytes, and linearity. The obtained parameters values confirm suitability of methodology to perform analysis of triterpenic compounds. Using the optimised and validated HPLC technique, four triterpenic compounds were separated and identified, and their specificity was confirmed. These compounds were corosolic acid, betulinic acid, oleanolic acid, and ursolic acid. Ursolic acid was the dominant compound in all the tested apple samples. The detected amount of betulinic acid was the lowest of all the identified triterpenic compounds. The greatest amounts of triterpenic compounds were detected in whole apple and apple peel samples of the 'Lodel' cultivar, and thus apples and apple extracts of this cultivar are potentially valuable for use in medical practice, for the prevention of various diseases, for adjunct therapy, for the isolation of individual compounds with a specific biological effect, and for the development and production of dietary supplements and functional food enriched in biologically active compounds. Acknowledgements. This work was supported by a grant from the Research Council of Lithuania, project No. MIP-17-8.

Keywords: apples, HPLC, triterpenic compounds, validation

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Authors: Pola Goldberg Oppenheimer, Stephan Hofmann, Sumeet Mahajan

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Owing to its fingerprint molecular specificity and high sensitivity, surface-enhanced Raman scattering (SERS) is an established analytical tool for chemical and biological sensing capable of single-molecule detection. A strong Raman signal can be generated from SERS-active platforms given the analyte is within the enhanced plasmon field generated near a noble-metal nanostructured substrate. The key requirement for generating strong plasmon resonances to provide this electromagnetic enhancement is an appropriate metal surface roughness. Controlling nanoscale features for generating these regions of high electromagnetic enhancement, the so-called SERS ‘hot-spots’, is still a challenge. Significant advances have been made in SERS research, with wide-ranging techniques to generate substrates with tunable size and shape of the nanoscale roughness features. Nevertheless, the development and application of SERS has been inhibited by the irreproducibility and complexity of fabrication routes. The ability to generate straightforward, cost-effective, multiplex-able and addressable SERS substrates with high enhancements is of profound interest for miniaturised sensing devices. Carbon nanotubes (CNTs) have been concurrently, a topic of extensive research however, their applications for plasmonics has been only recently beginning to gain interest. CNTs can provide low-cost, large-active-area patternable substrates which, coupled with appropriate functionalization capable to provide advanced SERS-platforms. Herein, advanced methods to generate CNT-based SERS active detection platforms will be discussed. First, a novel electrohydrodynamic (EHD) lithographic technique will be introduced for patterning CNT-polymer composites, providing a straightforward, single-step approach for generating high-fidelity sub-micron-sized nanocomposite structures within which anisotropic CNTs are vertically aligned. The created structures are readily fine-tuned, which is an important requirement for optimizing SERS to obtain the highest enhancements with each of the EHD-CNTs individual structural units functioning as an isolated sensor. Further, gold-functionalized VACNTFs are fabricated as SERS micro-platforms. The dependence on the VACNTs’ diameters and density play an important role in the Raman signal strength, thus highlighting the importance of structural parameters, previously overlooked in designing and fabricating optimized CNTs-based SERS nanoprobes. VACNTs forests patterned into predesigned pillar structures are further utilized for multiplex detection of bio-analytes. Since CNTs exhibit electrical conductivity and unique adsorption properties, these are further harnessed in the development of novel chemical and bio-sensing platforms.

Keywords: carbon nanotubes (CNTs), EHD patterning, SERS, vertically aligned carbon nanotube forests (VACNTF)

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