In this work, phosphorus-doped carbon dots (P-DESCDs) were successfully prepared using choline chloride/lactic acid type deep eutectic solvent and phosphoric acid as ingredients, and (3-aminopropyl) trimethoxysilane was used as a bridge to graft P-DESCDs onto the silica surface to obtain a new mixed-mode stationary phase (Sil-P-DESCDs) for reversed-phase and hydrophilic interaction liquid chromatography. The successful preparation of the stationary phase was confirmed by laser scanning confocal microscopy, elemental analysis, and Fourier transform infrared spectrometry. Interestingly, the doping of phosphorus greatly improved the separation performance and hydrophilicity of the Sil-P-DESCDs column. The Sil-P-DESCDs column was found to have certain hydrophobicity, hydrogen bonding ability and shape selectivity by Tanaka and Engelhardt standard test mixtures, and a series of hydrophilic and hydrophobic compounds such as alkylbenzenes, polycyclic aromatic hydrocarbons, sulfonamides, aromatic amines, phenols, flavonoids, nucleoside bases, and alkaloids. In addition, the effects of mobile phase ratio, column temperature, flow rate, salt concentration, and pH on the retention of analytes on Sil-P-DESCDs columns were investigated. Finally, the Sil-P-DESCDs column was applied to the qualitative and quantitative analysis of calcein-7-glucoside in the real sample of medicinal Astragalus pellets, and it was found at a concentration of 0.02 mg/mL.

The glycosylation status of proteins is increasingly used as biomarker to improve the reliability in the diagnosis and prognosis of diseases as relevant as cancer. This feeds the need for tools that allow its simple and reliable analysis and are compatible with applicability in the clinic. With this objective in mind, this work reports the first bioelectronic immunoplatforms described to date for the determination of glycosylated haptoglobin (Hp) and the simultaneous determination of total and glycosylated Hp. The bioelectronic immunoplatform is based on the implementation of non-competitive bioassays using two different antibodies or an antibody and a lectin on the surface of commercial magnetic microcarriers. The resulting bioconjugates are labeled with the horseradish peroxidase (HRP) enzyme, and after their magnetic capture on disposable electroplatforms, the amperometric transduction using the H2O2/hydroquinone (HQ) system allows the single or multiple detection. The developed immunoplatform achieves limits of detection (LODs) of 0.07 and 0.46 ng mL-1 for total and glycosylated Hp in buffer solution, respectively. The immunoplatform allows accurate determination using simple and relatively short protocols (approx. 75 min) of total and glycosylated Hp in the secretomes of in vitro-cultured colorectal cancer (CRC) cells with different metastatic potentials, which is not feasible, due to lack of sensitivity, by means of some commercial ELISA kits and Western blot methodology.

The short analysis time and constant temperature environment in the second dimension of two-dimensional comprehensive chromatography frequently causes wraparound problems, especially for complex high boilers. This problem can be solved by temperature programming on the second column, but since this requires heating and cooling the column in a matter of seconds, it is difficult to implement. In this study, we describe a method of accomplishing rapid heating and cooling with a resistively heated column cooled by compressed air. Critical to this method is minimizing the lag time between the actual temperature and the reported temperature by using the column heating element as the temperature sensor, virtually eliminating the danger of overshooting the temperature setpoint. This technique facilitates a ramp rate of up to 100 °C/s with minimal overshooting-well beyond the requirements of gas chromatography. A single-layer column bundle design allows a compressed-air cooling device to cool the column from 200 to 50 °C at an average rate of -21 °C/s. The secondary dimension temperature programming is facilitated by the longer secondary dimension time made possible by the direct flow modulation method. We evaluated the performance of the single-layer column bundle and demonstrated this method by applying it to a gasoline sample. We also compared this method with the traditional isothermal approach and found that use of the secondary temperature program reduced the naphthalene retention time from 12.1 to 6.3 s and its peak width at half height from 846 to 126 ms.

The objective of this work was to provide an unbiased comparison of one-dimensional reversed-phase liquid chromatography (1D-RPLC) and comprehensive two-dimensional RPLC (RPLC × RPLC), through calculations and experimental verifications. For this purpose, various quality descriptors were evaluated, including peak capacity, analysis time, dilution factor, number of runs in the second dimension, and injection volume. The same strategy was applied to small pharmaceuticals and peptides. Whatever the analysis time between 30 and 200 min, short columns of only 30 × 2.1 mm packed with sub-2-µm particles should be selected in both dimensions of the 2D-LC setup to obtain the best compromise in terms of peak capacity and sensitivity. The peak capacity in RPLC × RPLC vs. RPLC was significantly improved for analysis times beyond 5 min. However, extra-column volume located after the second-dimension column was found to be particularly critical for peptides, and up to 50% lower peak capacity was observed with MS vs. UV detection. Contrary to common belief, higher dilution is not always observed in RPLC × RPLC. With adequate analytical conditions, better sensitivity (in theory fivefold and in practice three- to fivefold) could be achieved in RPLC × RPLC compared to 1D-RPLC, regardless of the analysis time.

As fluorescence imaging in the second near-infrared window (NIR-II, 1000-1700 nm) has gained increasing attention, it is inevitable that NIR-II fluorophores, the cornerstone of NIR-II imaging, have come to the middle of the stage. NIR-II xanthenoid fluorophores with good stability, high brightness, and fluorescence adjustability are becoming popular. We here reviewed the recent progress of xanthenoid fluorophores with NIR-II emission for in vivo applications. Especially, we focus on the strategies used for longer wavelength and fluorescence regulation to construct OFF-ON or ratiometric NIR-II fluorescent probes.

Antibacterial polymer materials have gained interest due to their capability to inhibit or eradicate biofilms with greater efficiency in comparison with their monomeric counterparts. Among the antimicrobial and anti-biofouling polymers, catecholamine-based polymers - and in particular polydopamine - have been studied due to their favorable adhesion properties, which can be tuned by controlling the pH value. In this study, we used atomic force microscopy (AFM)-based spectroscopy to investigate the relation between the adhesion properties and surface charge density and the pH of electrochemically deposited polydopamine films presenting a dissociation constant of polydopamine of 6.3 ± 0.2 and a point of zero charge of 5.37 ± 0.06. Furthermore, using AFM and attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), the influence of the surface charge density of polydopamine on bacterial adhesion and biofilm formation was investigated. It was shown that the adhesion of Escherichia coli at positively charged polydopamine is three times higher compared to a negatively charged polymer, and that the formation of biofilms is favored at positively charged polymers.

A novel type of PEG-modified halloysite was prepared and used as a hydrophilic interaction and cation exchange mixed-mode sorbent for solid-phase extraction of biogenic amines in fish samples. The eluates were analyzed by high-performance liquid chromatography-ultraviolet detection after the derivatization with benzoyl chloride. The developed sorbent was characterized by scanning electron microscopy, infrared spectroscopy, X-ray diffraction, zeta potential analyzer, and thermo-gravimetric analysis. After the optimization of various parameters influencing the extraction efficiency, the PEG-modified halloysite-based SPE method was evaluated. The adsorption capacities of putrescine, spermine, phenethylamine, and histamine were as high as 9.3, 8.5, 5.7, and 5.6 mg g-1, respectively. Satisfactory reproducibility of sorbent preparation was obtained with within-batch and batch-to-batch relative standard deviations (RSDs) lower than 3.9% and 8.6%, respectively. The biogenic amine spiking recoveries in fish samples ranged from 84.3 to 105.5% with good RSDs lower than 7.8%. Intra-day and inter-day precision, expressed as RSDs, were better than 8.8%. The limits of detection of histamine, putrescine, phenethylamine, and spermine were 9.4, 1.9, 0.5, and 0.9 μg L-1, respectively. This work provides a new hydrophilic interaction and cation exchange mixed-mode sorbent and is successfully applied to the extraction of trace biogenic amines from fish samples.

The European pharmacopeia provides analytical methods for the chemical characterization of active pharmaceutical ingredients (APIs). However, the complexity of some APIs exceeds the limitations of the currently prevailing physicochemical methods. Sodium bituminosulfonate (SBS) is described by the collection of key parameters of generalizing criteria such as dry matter, sulfur and sodium content, and neutrality, but techniques to unravel the complexity on a molecular level are lacking. We present a study based on online derivatization with tetramethylammonium hydroxide in combination with comprehensive two-dimensional gas chromatography coupled to an electron ionization high-resolution time-of-flight mass spectrometer (GC × GC-HR-ToF-MS) for the chemical description of SBS as well as its process intermediates. The application of GC × GC allowed the comprehensive description of the chemical components in the API and the process intermediates for the first time. Furthermore, it was possible to classify peaks regarding their elemental and structural composition based on accurate mass information, elution behavior, and mass fragmentation pattern. This work demonstrates not only the general applicability, advantages but also limitations of GC × GC for the characterization of APIs for complex drugs.

An approach using pyrolysis with comprehensive two-dimensional gas chromatography with flame ionization detection is introduced for identifying common isolated plastic polymers. A quadrupole mass spectrometer is employed as a parallel detector to aid method development and improve polymer identification in complex matrices. Common plastic polymers including polyethylene, polypropylene, polystyrene, polyvinyl chloride, polyamide, poly(methyl methacrylate), styrene-butadiene rubber, and polyethylene terephthalate are accurately identified within a total analysis time of 45 min. A strategy to enhance compatibility of high-resolution capillary gas chromatography using a 150-µm internal diameter column technology and a larger internal volume microfurnace-based pyrolyzer is discussed. This strategy resulted in minimizing the band broadening effect caused by the pyrolyzer's internal volume and overcoming the slow pressure buildup when the sample is inserted into the furnace. Prolonged pressure buildup to reach a final pressure setting can cause a safety shutdown to the pneumatic control system. The developed approach is complementary to spectroscopic techniques by offering mass based, chemical composition analysis of plastics.

Food supplement authentication is an important concern worldwide due to the ascending consumption related to health benefits and its lack of effective regulation in underdeveloped countries, making it a target of fraudulent activities. In this context, this study evaluated fish oil supplements by comprehensive two-dimensional gas chromatography coupled to mass spectrometry (GC×GC-MS) to obtain fingerprints, which were used to build predictive models for automated authentication of the most popular products sold in Brazil. The authentication process relied on a one-class classifier model using data-driven soft independent modeling of class analogy (DD-SIMCA). The output of the model was a binary classifier: certified IFOS fish oils and non-certified ones - regardless of the source of adulteration. The compositional analysis showed a significant variation in the samples, which validated the need for reliable statistical models. The DD-SIMCA algorithm is still incipient in GC×GC studies, but it proved to be an excellent tool for authenticity purposes, achieving a chemometric model with a sensitivity of 100%, specificity of 98.6%, and accuracy of 99.0% for fish oil authentication. Finally, orthogonalized partial least square discriminant analysis (OPLS-DA) was used to identify the features that distinguished the groups, which ascertained the results of the DD-SIMCA model that IFOS-certified oils are positively correlated to omega-3 fatty acids, including eicosapentaenoic acid (EPA, C20:5 n-3) and docosahexaenoic acid (DHA, C22:6 n-3).

This work describes a novel automated and rapid method for bottom-up proteomics combining protein isolation with a micro-immobilised enzyme reactor (IMER). Crosslinking chemistry based on 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling was exploited to immobilise trypsin and antibodies onto customisable silica particles coated with carboxymethylated dextran (CMD). This novel silica-CMD solid-phase extraction material was characterised using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), conductometric titrations and enzymatic colorimetric assays. Micro-solid-phase extraction (μSPE) cartridges equipped with the modified CMD material were employed and integrated into an automated and repeatable workflow using a sample preparation workstation to achieve rapid and repeatable protein isolation and pre-concentration, followed by tryptic digestion producing peptide fragments that were identified by liquid chromatography mass spectrometry (LC-MS).