5 figures.-- Supporting information available., The question of whether exosome lipids can be considered as potential cancer biomarkers faces our current limited knowledge of their composition. This is due to the difficulty in isolating pure exosomes, the variability of the biological sources from which they are extracted, and the uncertainty of the methods for lipid characterization. Here, we present a procedure to isolate exosomes and obtain a deep, repeatable, and rapid phospholipid (PL) composition of their lipid extracts, from embryonic murine fibroblasts (NIH-3T3 cell line) and none (B16-F1) and high (B16-F10) metastatic murine skin melanoma cells. The analytical method is based on High Performance Thin-Layer Chromatography with Ultraviolet and fluorescence densitometry and coupled to Electrospray (ESI)-tandem Mass Spectrometry (MS). Under the conditions described in this work, separation and determination of PL classes, (sphingomyelins, SM; phosphatidylcholines, PC; phosphatidylserines, PS; and phosphatidylethanolamines, PE) were achieved, expressed as µg PL/100 µg exosome protein, obtained by bicinchoninic acid assay (BCA). A detailed structural characterization of molecular species of each PL class was performed by simultaneous positive and negative ESI-MS and MS/MS directly from the chromatographic plate, thanks to an elution-based interface., Financial support for this project was provided by DGA-FEDER (E25_20R, N&SB), AES
PI21/00036, CSIC 202180E076, ISCIII PI19/01007, DGA E25-20R, and ERC Advanced Grant CADENCE (grant No. ERC-2016-ADG-742684)., Peer reviewed

5 figures.-- Supplementary information available., Heterostructures of two-dimensional (2D) materials using graphene and MoS2 have enabled both pivotal fundamental studies and unprecedented sensing properties. These heterosystems are intriguing when graphene and MoS2 are interfaced with 2D sheets that emulate biomolecules, such as amino-terminated oligoglycine self-assemblies (known as tectomers). The adsorption of tectomer sheets over graphene and MoS2 modulates the physicochemical properties through electronic charge migration and mechanical stress transfer. Here, we present a systematic study by Raman spectroscopy and tectomer-functionalised scanning probe microscopy to understand mechanical strain, charge transfer and binding affinity in tectomer/graphene and tectomer/MoS2 hybrid structures. Raman mapping reveals distinctive thickness dependence of tectomer-induced charge transfer to MoS2, showing p-doping on monolayer MoS2 and n-doping on multilayer MoS2. By contrast, graphene is n-doped by tectomer independently of layer number, as confirmed by x-ray photoelectron spectroscopy. The interfacial adhesion between the amino groups and 2D materials are further explored using tectomer-functionalised probe microscopy. It is demonstrated here that these probes have potential for chemically sensitive imaging of 2D materials, which will be useful for mapping chemically distinct domains of surfaces and the number of layers. The facile tectomer-coating approach described here is an attractive soft-chemistry strategy for high-density amine-functionalisation of atomic force microscopy probes, therefore opening promising avenues for sensor applications., We would like to acknowledge strategic development funding from the University of Sussex. Additional funding from the Aragon Government is also acknowledged (Grupo de Nanosensores y Sistemas Bioanalíticos (N&SB), ref. E25_20R). [...] This work was also partially supported by a UKRI Future Leaders Fellowship (Grant No. MR/T042664/1)., Peer reviewed

8 figures.-- Supplementary information available., A variety of two-dimensional (2D) nanomaterials, including graphene oxide and clays, are known to stabilize Pickering emulsions to fabricate structures for functions in sensors, catalysts, and encapsulation. We introduce here a novel Pickering emulsion using self-assembled amphiphilic triblock oligoglycine as the emulsifier. Peptide amphiphiles are more responsive to environmental changes (e.g., pH, temperature, and ionic strength) than inorganic 2D materials, which have a chemically rigid, in-plane structure. Noncovalent forces between the peptide molecules change with the environment, thereby imparting responsiveness. We provide new evidence that the biantennary oligoglycine, Gly4–NH–C10H20–NH–Gly4, self-assembles into 2D platelet structures, denoted as tectomers, in solution at a neutral buffered pH using small-angle X-ray scattering and molecular dynamics simulations. The molecules are stacked in the platelets with a linear conformation, rather than in a U-shape. We discovered that the lamellar oligoglycine platelets adsorbed at an oil/water interface and stabilized oil-in-water emulsions. This is the first report of 2D oligoglycine platelets being used as a Pickering stabilizer. The emulsions showed a strong pH response in an acidic environment. Thus, upon reducing the pH, the protonation of the terminal amino groups of the oligoglycine induced disassembly of the lamellar structure due to repulsive electrostatic forces, leading to emulsion destabilization. To demonstrate the application of the material, we show that a model active ingredient, β-carotene, in the oil is released upon decreasing the pH. Interestingly, in pH 9 buffer, the morphology of the oil droplets evolved over time, as the oligoglycine stabilizer created progressively a thicker interfacial layer. This demonstration opens a new route to use self-assembled synthetic peptide amphiphiles to stabilize Pickering emulsions, which can be significant for biomedical and pharmaceutical applications., Z.H. acknowledges the Chinese Scholarship Council (CSC 201906950049) and the Vice-Chancellor’s Scholarship Fund for providing a PhD studentship. E.C. acknowledges the Faculty of Science and Engineering at the University of Groningen for PhD funding. [...] This research was supported by the Aragón Regional Government (project E25_20R) [...]., Peer reviewed

Analysis of the GISAXS data:
The GISAXS pattern was further analyzed with the aim to estimate the thickness of the regions composing the lamellar structure.
We recall that the SAXS intensity can be well described by the general equation:
𝐼(𝑞)=Δρ2𝑃(𝑞)𝑆(𝑞) (S1)
where Δρ2 is the contrast term defined by the electron density difference within the system, 𝑃(𝑞) is the so-called form factor, describing the scattered intensity from the objects of a certain shape and size, and 𝑆(𝑞) is the structure factor, describing the spatial correlation between adjacent scattering objects.
The many reflections observed in the GISAXS pattern (Figure 4c) are due to the presence of a multilayer structure, generated by the alternating repetition of the peptide and alkyl layers assembled in a stacked fashion (Figure 4d). The absence of the 5th diffraction reflection is directly related to the thickness of the layers. Since the system is molecularly well-defined, destructive interference between the minimum of the lamellar form factor and the 5th reflection maxima occurs. To confirm this, in Figure S6 we present the simulated intensity for the form factor (𝑃(𝑞)) of a layer with thickness h = 0.83 nm and the structure factor (𝑆(𝑞)) for a multilayered structure with periodicity d = 4.19 nm. For completeness, the simulated background and the total fitted curve are also included in the figure. In these simulations, the scattering from a homogeneous layer with thickness, h, and a lateral size larger than 1000 nm
was used, while the structure factor using paracrystalline disorder was used.1 The simulations and fitting were performed using MATLAB.
It can be clearly seen that the minimum of the form factor cancels the maximum of the
structure factor. Assuming that the system is two phase, the thickness of the second phase must be 4.19 nm - 0.83 nm = 3.36 nm.
We note that the mismatch in the relative peak height is most probably due to the texture
of the sample, as the fitted data have been extracted from the vertical cut along the qz direction of the GISAXS patterns.
Pendant Drop Interfacial Tensiometry:
A drop shape analyser (Krüss, FTA DSA1000B) was used for pendant drop tensiometry to measure the interfacial tension at the interface between the oil and different aqueous solutions. In these experiments, the aqueous phase was either DI-water, pH 7.0 buffer, 0.5 mg/mL 2T solutions in DI-water, or 0.5 mg/mL 2T in pH 7.0 buffer solution. A drop of the aqueous phase with a volume of approximately 7 μL was injected and hung from a 0.5 mm diameter needle in the less dense oil phase. Images of the drop shape were recorded using the video camera system on the apparatus and analyzed (using FTA32 software) to obtain the interfacial tension value over time. The experiments were conducted in a temperature-controlled room with a temperature of 22 +/-1 °C.
The forces that determine the shape of the pendant drop are mainly the balance of surface tension, buoyancy force and gravitation. The surface tension seeks to minimize the surface area and make the drop take a spherical shape. The difference of gravitation and buoyancy force, on the other hand, stretches the drop from this spherical shape and a typical pear-like shape results. The interfacial tension for a drop with a diameter of 𝐷𝑒𝑞 at its equator was calculated as:
𝛾=𝛥𝜌·𝑔𝐷𝑒𝑞2𝐻 (S2)
where 𝛥𝜌 is the density difference between water and oil, 𝑔 is the acceleration due to gravity (taken as 9.8 m/s2), and 𝐻 is a shape factor that is decided by the shape of the drop. Here, the water density was taken to be ρw = 0.998 g/cm3 (obtained from the FTA32 software), and the density of sunflower oil3 was taken as ρo = 0.918 g/cm3. Six replicate measurements were performed on the sunflower oil/DI water interface. The standard deviation was 0.4 mN/m.
In pH 7.0 buffer solutions, there is a self-assembly of 2T platelets. The platelets are large enough to be subject to gravitational effects, which made the interfacial tension measurements unreliable.-- Licensed under CC-BY 4.0., Size, SAXS, and microscope images of 2T in pH 7.0 buffer, water, and pH 4.0 buffer; description of coarse-grained simulations; molecular model; photograph of the 2T films from the oil/water interface; Raman spectrum of pure sunflower oil; interfacial tension measurements; aging of the emulsion; destabilization of a β-carotene emulsion; SEM image of the spin-coated emulsion; and EDS measurements of the elements in a dried 2T-stabilized oil drop (PDF)
Coarse-grained simulation of the peptide in water, demonstrating the formation process of the 2D peptide assembly (MP4)
All-atom simulation of 2T attaching to the oil/water interface (MP4), Z.H. acknowledges the Chinese Scholarship Council (CSC 201906950049) and the Vice-Chancellor’s Scholarship Fund for providing a PhD studentship. E.C. acknowledges the Faculty of Science and Engineering at the University of Groningen for PhD funding. [...] This research was supported by the Aragón Regional Government (project E25_20R) [...]., Peer reviewed

Carbon nanomaterials have attracted increasing attention in biomedicine recently to be used as drug nanocarriers suitable for medical treatments, due to their large surface area, high cellular internalization and preferential tumor accumulation, that enable these nanomaterials to transport chemotherapeutic agents preferentially to tumor sites, thereby reducing drug toxic side effects. However, there are widespread concerns on the inherent cytotoxicity of carbon nanomaterials, which remains controversial to this day, with studies demonstrating conflicting results. We investigated here in vitro toxicity of various carbon nanomaterials in human epithelial colorectal adenocarcinoma (Caco-2) cells and human breast adenocarcinoma (MCF-7) cells. Carbon nanohorns (CNH), carbon nanotubes (CNT), carbon nanoplatelets (CNP), graphene oxide (GO), reduced graphene oxide (GO) and nanodiamonds (ND) were systematically compared, using Pluronic F-127 dispersant. Cell viability after carbon nanomaterial treatment followed the order CNP < CNH < RGO < CNT < GO < ND, being the effect more pronounced on the more rapidly dividing Caco-2 cells. CNP produced remarkably high reactive oxygen species (ROS) levels. Furthermore, the potential of these materials as nanocarriers in the field of drug delivery of doxorubicin and camptothecin anticancer drugs was also compared. In all cases the carbon nanomaterial/drug complexes resulted in improved anticancer activity compared to that of the free drug, being the efficiency largely dependent of the carbon nanomaterial hydrophobicity and surface chemistry. These fundamental studies are of paramount importance as screening and risk-to-benefit assessment towards the development of smart carbon nanomaterial-based nanocarriers.

The question of whether exosome lipids can be considered as potential cancer biomarkers faces our current limited knowledge of their composition. This is due to the difficulty in isolating pure exosomes, the variability of the biological sources from which they are extracted, and the uncertainty of the methods for lipid characterization. Here, we present a procedure to isolate exosomes and obtain a deep, repeatable, and rapid phospholipid (PL) composition of their lipid extracts, from embryonic murine fibroblasts (NIH-3T3 cell line) and none (B16-F1) and high (B16-F10) metastatic murine skin melanoma cells. The analytical method is based on High Performance Thin-Layer Chromatography with Ultraviolet and fluorescence densitometry and coupled to Electrospray (ESI)-tandem Mass Spectrometry (MS). Under the conditions described in this work, separation and determination of PL classes, (sphingomyelins, SM; phosphatidylcholines, PC; phosphatidylserines, PS; and phosphatidylethanolamines, PE) were achieved, expressed as mu g PL/100 mu g exosome protein, obtained by bicinchoninic acid assay (BCA). A detailed structural characterization of molecular species of each PL class was performed by simultaneous positive and negative ESI-MS and MS/MS directly from the chromatographic plate, thanks to an elution-based interface.

An enzymatic-colorimetric method has been developed based on the reaction between l-phenylalanine (l-Phe) and the l-amino acid oxidase (LAAO) in the presence of Au(III), which has led to the formation of gold nanoparticles. The intensity of the localized surface plasmon resonance (LSPR) band of the generated nanoparticles (550 nm) can be related to the concentration of l-Phe in the sample. The mechanism of the LAAO-l-Phe enzyme reaction in the presence of Au(III) has been studied through the evaluation and optimization of experimental conditions. These studies have reinforced the hypothesis that the catalytic center of the enzyme helps the Au(III) reduction and, thanks to the protein, the Au0 form is stabilized as gold nanoparticles (AuNPs). In the calibration study, a sigmoidal relationship between the concentration of the substrate and the LSPR of the nanoparticles was observed. The linearization of the signal has allowed the determination of l-Phe in the range from 17 to 500 µM with an RSD% (150 μM) of 4.8% (n = 3). The method is free of other amino acid interference normally found in blood plasma. These highly competitive results open the possibility of further development of a rapid method for l-Phe determination based on colorimetry.

In the past twelve years, digital image colorimetry (DIC) on smartphones has acquired great importance as an alternative to the most common analytical techniques. This analysis method is based on fast, low-cost, and easily-accessible technology, which can provide quantitative information about an analyte through the color changes of a digital image. Despite the fact that DIC is very widespread, it is not exempt from a series of problems that are not fully resolved yet, such as variability of the measurements between smartphones, image format in which color information is stored, power distribution of the illuminant used for the measurements, among others. This article proposes a methodology for the standardization and correction of these problems using self-developed software, together with the use of a 3D printed light box. This methodology is applied to three different colorimetric analyses using different types and brands of smartphones, proving that comparable measurements between devices can be achieved. As color can be related to many target analytes, establishing this measurement methodology can lead to new control analysis applicable to diverse sectors such as alimentary, industrial, agrarian, or sanitary.

Bacteroides fragilis is an abundant commensal component of the healthy human colon. However, under dysbiotic conditions, enterotoxigenic B. fragilis (ETBF) may arise and elicit diarrhea, anaerobic bacteremia, inflammatory bowel disease, and colorectal cancer. Most worrisome, ETBF is resistant to many disparate antibiotics. ETBF's only recognized specific virulence factor is a zinc-dependent metallopeptidase (MP) called B. fragilis toxin (BFT) or fragilysin, which damages the intestinal mucosa and triggers disease-related signaling mechanisms. Thus, therapeutic targeting of BFT is expected to limit ETBF pathogenicity and improve the prognosis for patients. We focused on one of the naturally occurring BFT isoforms, BFT-3, and managed to repurpose several approved drugs as BFT-3 inhibitors through a combination of biophysical, biochemical, structural, and cellular techniques. In contrast to canonical MP inhibitors, which target the active site of mature enzymes, these effectors bind to a distal allosteric site in the proBFT-3 zymogen structure, which stabilizes a partially unstructured, zinc-free enzyme conformation by shifting a zinc-dependent disorder-to-order equilibrium. This yields proBTF-3 incompetent for autoactivation, thus ablating hydrolytic activity of the mature toxin. Additionally, a similar destabilizing effect is observed for the activated protease according to biophysical and biochemical data. Our strategy paves a novel way for the development of highly specific inhibitors of ETBF-mediated enteropathogenic conditions.

A variety of two-dimensional (2D) nanomaterials, including graphene oxide and clays, are known to stabilize Pickering emulsions to fabricate structures for functions in sensors, catalysts, and encapsulation. We introduce here a novel Pickering emulsion using self-assembled amphiphilic triblock oligoglycine as the emulsifier. Peptide amphiphiles are more responsive to environmental changes (e.g., pH, temperature, and ionic strength) than inorganic 2D materials, which have a chemically rigid, in-plane structure. Noncovalent forces between the peptide molecules change with the environment, thereby imparting responsiveness. We provide new evidence that the biantennary oligoglycine, Gly4–NH–C10H20–NH–Gly4, self-assembles into 2D platelet structures, denoted as tectomers, in solution at a neutral buffered pH using small-angle X-ray scattering and molecular dynamics simulations. The molecules are stacked in the platelets with a linear conformation, rather than in a U-shape. We discovered that the lamellar oligoglycine platelets adsorbed at an oil/water interface and stabilized oil-in-water emulsions. This is the first report of 2D oligoglycine platelets being used as a Pickering stabilizer. The emulsions showed a strong pH response in an acidic environment. Thus, upon reducing the pH, the protonation of the terminal amino groups of the oligoglycine induced disassembly of the lamellar structure due to repulsive electrostatic forces, leading to emulsion destabilization. To demonstrate the application of the material, we show that a model active ingredient, β-carotene, in the oil is released upon decreasing the pH. Interestingly, in pH 9 buffer, the morphology of the oil droplets evolved over time, as the oligoglycine stabilizer created progressively a thicker interfacial layer. This demonstration opens a new route to use self-assembled synthetic peptide amphiphiles to stabilize Pickering emulsions, which can be significant for biomedical and pharmaceutical applications.

Heterostructures of two-dimensional (2D) materials using graphene and MoS2 have enabled both pivotal fundamental studies and unprecedented sensing properties. These heterosystems are intriguing when graphene and MoS2 are interfaced with 2D sheets that emulate biomolecules, such as amino-terminated oligoglycine self-assemblies (known as tectomers). The adsorption of tectomer sheets over graphene and MoS2 modulates the physicochemical properties through electronic charge migration and mechanical stress transfer. Here, we present a systematic study by Raman spectroscopy and tectomer-functionalised scanning probe microscopy to understand mechanical strain, charge transfer and binding affinity in tectomer/graphene and tectomer/MoS2 hybrid structures. Raman mapping reveals distinctive thickness dependence of tectomer-induced charge transfer to MoS2, showing p-doping on monolayer MoS2 and n-doping on multilayer MoS2. By contrast, graphene is n-doped by tectomer independently of layer number, as confirmed by x-ray photoelectron spectroscopy. The interfacial adhesion between the amino groups and 2D materials are further explored using tectomer-functionalised probe microscopy. It is demonstrated here that these probes have potential for chemically sensitive imaging of 2D materials, which will be useful for mapping chemically distinct domains of surfaces and the number of layers. The facile tectomer-coating approach described here is an attractive soft-chemistry strategy for high-density amine-functionalisation of atomic force microscopy probes, therefore opening promising avenues for sensor applications.

Hypothesis: Surfactants in emulsions sometimes do not provide adequate stability against coalescence, whereas Pickering emulsions often offer greater stability. In a search for stabilizers offering biocompatibility, we hypothesized that carboxylated nanodiamonds (ND) would impart stability to Pickering emulsions. Experiments: We successfully prepared Pickering emulsions of sunflower oil in water via two different methods: membrane emulsification and probe sonication. The first method was only possible when the pH of the aqueous ND suspension was ≤ 4. Findings: Pendant-drop tensiometry confirmed that carboxylated ND is adsorbed at the oil/water interface, with a greater decrease in interfacial tension found with increasing ND concentrations in the aqueous phase. The carboxylated ND become more hydrophilic with increasing pH, according to three-phase contact angle analysis, because of deprotonation of the carboxylic acid groups. Membrane emulsification yielded larger (about 30 µm) oil droplets, probe sonication produced smaller (sub-μm) oil droplets. The Pickering emulsions show high stability against mechanical vibration and long-term storage for one year. They remain stable against coalescence across a wide range of pH values. Sonicated emulsions show stability against creaming. In this first-ever systematic study of carboxylated ND-stabilized Pickering emulsions, we demonstrate a promising application in the delivery of β-carotene, as a model active ingredient.

Tyramine oxidase (TAO), peroxidase (HRP), and Amplex Red (AR) have been immobilized on cellulose to obtain disposable biosensors for the determination of histamine. During the enzymatic reaction, AR is oxidized and a pink spot is obtained. Using a smartphone and measuring the G (green) color coordinate, histamine can be determined in the presence of other biogenic amines (putrescine and cadaverine) in concentrations ranging from 2·10−5 M to 5·10−4 M with a 7.5·10−6 M limit of detection (LoD). Despite tyramine interference, experimental conditions are provided which allow rapid and simple histamine and simultaneous histamine/tyramine (semi)quantitative determination in mixtures. Finally, tyramine and histamine were determined in a tuna extract with good results (compared to the reference HPLC–MS method). The methodology can also be applied in solution allowing histamine (and simultaneous histamine/tyramine) determination with a lower LoD (1.8·10−7 M) and a similar selectivity.

This work details the enzymatic generation of fluorescence nanomaterials and the use of this optical signal as the analytical parameter for the quantification of the substrate. More specifically, fluorescent copper nanoclusters have been obtained during the enzymatic reaction of tyramine oxidase and tyramine in the presence of Cu(II); the fluorescence intensity being proportional to the concentration of tyramine. The nanoclusters obtained show fluorescence at 445 nm by being excited at 320 nm and have been characterized by TEM, EDX, and XPS. The formation mechanism has also been studied, suggesting that under the optimal conditions (0.1 M MES buffer and pH = 6), the formation of the nanoclusters is due to the reducing properties of the product of the enzymatic reaction (p-hydroxybenzaldehyde) in MES buffer. The method shows a linear relationship with the concentration of tyramine in the range from 1.0·10−5 to 2.5·10−4 M, a RSD of 3% (n = 5) and a LOD of 6.3·10−6 M. The method has been applied to the determination of tyramine in sausage with good results.

The development of optical sensors for in situ testing has become of great interest in the rapid diagnostics industry. We report here the development of simple, low-cost optical nanosensors for the semi-quantitative detection or naked-eye detection of tyramine (a biogenic amine whose production is commonly associated with food spoilage) when coupled to Au(III)/tectomer films deposited on polylactic acid (PLA) supports. Tectomers are two-dimensional oligoglycine self-assemblies, whose terminal amino groups enable both the immobilization of Au(III) and its adhesion to PLA. Upon exposure to tyramine, a non-enzymatic redox reaction takes place in which Au(III) in the tectomer matrix is reduced by tyramine to gold nanoparticles, whose reddish-purple color depends on the tyramine concentration and can be identified by measuring the RGB coordinates (Red–Green–Blue coordinates) using a smartphone color recognition app. Moreover, a more accurate quantification of tyramine in the range from 0.048 to 10 μM could be performed by measuring the reflectance of the sensing layers and the absorbance of the characteristic 550 nm plasmon band of the gold nanoparticles. The relative standard deviation (RSD) of the method was 4.2% (n = 5) with a limit of detection (LOD) of 0.014 μM. A remarkable selectivity was achieved for tyramine detection in the presence of other biogenic amines, especially histamine. This methodology, based on the optical properties of Au(III)/tectomer hybrid coatings, is promising for its application in food quality control and smart food packaging.

In situ enzymatic generation of bimetallic nanoparticles, mainly Au/Pt, overcomes the drawbacks (continuous absorbance drift, modest LOQ, and long-time reaction) observed when AuNP alone are produced. In this study, Au/Pt nanoparticles have been characterized by EDS, XPS, and HRTEM images using the enzymatic determination of tyramine with tyramine oxidase (TAO) as a model. Under experimental conditions, the Au/Pt NPs show an absorption maximum at 580 nm which can be related to the concentration of tyramine in the range 1.0 × 10-6M to 2.5 × 10-4M with a RSD of 3.4% (n = 5, using 5 × 10-6M tyramine). The Au/Pt system enables low LOQ (1.0 × 10−6 M), high reduction of the absorbance drift, and a significant shortening of the reaction time (i.e., from 30 to 2 min for a [tyramine] = 1 × 10−4M); additionally, a better selectivity is also obtained. The method has been applied to tyramine determination in cured cheese and no significant differences were obtained compared to a reference method (HRP:TMB). The effect of Pt(II) seems to involve the previous reduction of Au(III) to Au(I) and NP generation from this oxidation state. Finally, a three-step (nucleation-growth-aggregation) kinetic model for the generation of NPs is proposed; this has enabled us to obtain a mathematical equation which explains the experimentally observed variation of the absorbance with time.

New β-diketonate platinum (II) complexes, containing a cyclometalated N-heterocyclic carbene [Pt(Naph^C*iPr)(acac)] (3A) (HNaph^C*-κC* = 3-isopropyl-1-(naphthalen-2-yl)-1H-imidazole-2-ylidene), or a cyclometalated pyrazole, [Pt(Naph^Npz)(acac)] (3B) (HC^Npz = 1-(naphthalen-2-yl)-1H-pyrazole) and [Pt(Naph^Ndmpz)(acac)] (3B') (HC^Ndmpz = 1-(naphthalen-2-yl)-1H-3,5-dimethylpyrazole) have been prepared and characterized. Their absorption and emission properties in films of ethyl cellulose (EC) were determined along with those of the already reported for complex [Pt(Naph^C*Me)(acac)] (3A'). They showed that all four β-diketonate complexes display a bright phosphorescent emission with maxima in the blue region (λmax ∼ 480 nm for 3A and 3A'; 490 nm 3B and 3B'). The higher quantum yield (QY), longer decay times and greater oxygen sensitivity were exhibited by the Naph^C* derivatives, compared to the Naph^N ones. Polyacrylamide membranes with entrapped 3A' as dye, and glucose oxidase (GOx) enzyme were used for monitoring glucose level. The RSD is about 5% and the detection limit is at ∼5·10−4 M, with a response time usually of 10–15 min working in stop-flow mode. These platinum-based membranes respond reversibly to glucose for, at least, 20 measures. 3A’ is the first Pt(II) complex bearing a cyclometalated N-heterocyclic carbene ever used as dye for sensing glucose.

The last triennium has shown a significant contribution of high-performance thin-layer chromatography (HPTLC) to lipidomics. HPTLC separation in combination with radio- and/or ultraviolet-fluorescence (UV-FL) densitometry was the technique of choice for tracking the transport of phospholipids among different cellular compartments in a number of biological systems, e.g., Gram-negative bacteria, yeast membranes, endoplasmic reticulum/mitochondrial membrane interface, and also to monitor the lipid transfer activity of a protein. Likewise, a significant number of HPTLC methods were developed to determine variations in the content of different lipid classes and subclasses after using genetic knockouts of cells. Radiolabeled control cells and cells whose genes were deleted using genomic editing allowed to study by HPTLC the effect of the disruption of the related proteins on the corresponding biosynthetic pathways. As well, direct interface-based coupling of HPTLC to mass spectrometry (MS) using mostly electrospray ionization(ESI) and a variety of mass analyzers, has gained new momentum in this period. Obtaining a large amount of information online in a very short time from complex biological samples, and the structural identification of target and non-target, unknown lipids, make this technique a useful tool for lipidomics. Quantitative issues related to HPTLC-MS are also discussed in this work.

The characterization and counting of foreign bodies and impurities in flour and semolina from wheat are decisive analytical parameters of safety and quality which should be evaluated in the context of efficiency and low cost. Moreover, their recognition and counting are required by the regulatory and quality norms of the sector. International standards such as UNI 10941:2001 could be applied but this has subjective interpretation. In this paper, a new analytical semi-automatic method based on digital images is assessed and proposed. The main features of the method are based on the use of representative sample images that could be taken under controlled illumination conditions with a smartphone. The images were then analyzed using a macro-script adapted to the free software Fiji-ImageJ for image processing. By changing the image color format to grey and its contrast, a threshold intensity, or cutoff, was selected to distinguish foreign bodies from the rest of the product. And the number of particles was counted. Finally, four different fractions of components in the product were recognized which characterized the type of product. The estimated processing time was less than 60 s. The method has been validated against 14 reference samples that were previously studied using the standard UNI 10941:2001. These samples presented low, medium and high particle content, as well as different background colors of the matrix product, from white to yellow. The results were obtained with an average of 6 ROIs taken in different locations of the same digital image. Figures of merit of the procedure such as the biases presented relative differences of less than +/- 20%, against reference values in the worst case. The reproducibility of the measurements, examining different locations, is better than 30-40% RSD. Likewise, the reproducibility in the same ROI but with short scans in the threshold of selection produced response ranges of less than 30% RSD. These parameters are consistent with those prevalent in the sector and this type of product.

In this manuscript a method for the fluorometric determination of tyramine is described. It is based on the direct reaction between Au(III) and tyramine in a phosphate buffer which produces fluorescent gold nanoclusters (AuNC) (λexc = 320 nm, λem = 410 nm) with a diameter of 1.50 ± 0.06 nm. The Au(III) and buffer solutions are mixed and after 140 s, tyramine solution is added; which produces a fast and stable fluorescence signal. The formation of AuNC is demonstrated by STEM and, more importantly, this reaction could be followed by Atomic Fluorescence Microscopy (AFM). The method allows the determination of tyramine in the range from 6.0x10−7 M (limit of quantification) up to 1.2x10−4 M; with a relative standard deviation (RSD) ranges from 1.8% to 4.4% depending on the tyramine concentration. The mechanism of AuNC formation involves the Au(III) reduction via the phenol group and the complexation with the amine group. Putrescine and cadaverine do not produce interference, meanwhile histamine causes a proportional decrease in the signal which can be overcome by the standard addition method. The method was applied to the determination of tyramine in a tuna and cheese samples and the results obtained are in statistical agreement with these obtained using a validated or standard method.

The development of membranes for nanofiltration applications requires not only selective layers but also suitable supports to control their synthesis as well as to enable efficient and competitive membrane performances. Single-walled carbon nanotube free-standing films (denoted as buckypaper) have been used as supports for the preparation of polyamide (PA) layers by interfacial polymerization (IP) and tested for dead-end nanofiltration for dyes removal (265–1017 Da) from water and organic solvents. The arrangement of the phases during the IP is essential, thus impregnation on the buckypaper support, first with the organic phase and then with the aqueous phase (denoted as inverse IP, iIP) leads to permeances (of up to 28.0 and 31.4 L m-2 h-1 bar-1 for water and methanol, respectively) and rejection values (>96%) that exceed those of the membranes prepared by reversing the impregnation order. Secondly, a double layer PA/zeolitic imidazolate framework (ZIF-8 or ZIF-93) was prepared on the buckypaper (bp). The design of this double layer led to superior membrane performance, in particular for the hydrophilic ZIF-93, as it changes the PA layer properties (increasing both hydrophilicity and surface roughness) providing higher permeances (up to 59.3 and 76.0 L m-2 h-1 bar-1 for water and methanol, respectively) and dye rejections (>98.5%) than the bare PA layer prepared by iIP. This attractive performance of the PA/ZIF-93/bp membrane has been corroborated with experiments in cross-flow nanofiltration and dead-end nanofiltration of aqueous salt solutions, long-term stability nanofiltration and the study the membrane separation performance after a chlorine treatment. The results reported here therefore show the enormous potential of these membrane architectures for a variety of selective separation processes. © 2022 The Authors