Separations, Volume 9, Issue 5 (May 2022) – 31 articles

The gas chromatography–mass spectrometry (GC-MS) approach is established for the simultaneous determination of fenchone (FCO) and trans-anethole (TOH) in the essential oils and methanolic extracts of fennel (Foeniculum vulgare Mill.) fruits obtained from India (IND), Pakistan (PAK), and Saudi Arabia (SA). The simultaneous determination of FCO and TOH was performed via Agilent 190914S HP–5MS fused-silica capillary column (30 m × 250 µm ID, 0.25 µm film thickness). The proposed GC-MS approach was linear in the range of 0.10–50 µg/g for FCO and TOH. FCO’s detection (LOD) and quantification (LOQ) limits were calculated to be 0.04 and 0.12 µg/g, respectively. The LOD and LOQ values for TOH, on the other hand, were calculated to be 0.05 and 0.15 µg/g, respectively. In addition, the proposed GC-MS approach was accurate and precise for the simultaneous determination of FCO and TOH. The amount of FCO in essential oils of F. vulgare was computed as 0.021, 0.034, and 0.029 mg/g in the samples obtained from IND, PAK, and SA, respectively. The amount of TOH in the essential oils of F. vulgare was computed as 7.40, 14.8, and 10.2 mg/g in the samples obtained from IND, PAK, and SA, respectively. However, the amount of FCO in the methanolic extract of F. vulgare was estimated as 0.031, 0.021, and 0.057 mg/g in the samples obtained from IND, PAK, and SA, respectively. On the other hand, the amount of TOH in the methanolic extract of F. vulgare was estimated as 0.440, 0.498, and 1.74 mg/g in the samples obtained from IND, PAK, and SA, respectively. These findings suggested that the proposed GC-MS approach might be used to simultaneously determine the FCO and TOH in a variety of essential oils and plant extracts. Full article

The material composition of a group of Rodovetri, hand-painted animation cels, made in the 1950s and 1960s for Italian television, has been determined by Flash Py-GC-MS, Thermally assisted Hydrolysis and Methylation-(Py-)GC-MS and GC-MS investigations, where, for the pyrolysis applications, the pyrolyzer is connected septum-less to the GC inlet. The condition of the selected animation cels was generally poor (yellowing, exudation, warping, cracking). The cels are made of plasticized cellulose acetate (CA) and decorated with alkyd paint. Exudating plasticizers from the CA support migrated into the paint; this has softened the paint layers and made the cels stick together. CA is known to be very unstable and easily subjected to degradation, which cannot be reversed, only slowed down by preventive conservation. Most of the cels are plasticized with bis(2-methoxyethyl) phthalate (DMEP) and triphenyl phosphate (TPP). The research shows a relationship between the degradation of CA and the presence of TPP and DMEP. Full article

The effective purification of corrosive gases at the cathode air stream side is essential for proton exchange membrane fuel cells’ performance in real-world applications. Gas molecular diffusion depth along the pore channel is a sufficient parameter that determines the effectiveness of the porous purification media. The collision between gas molecules and pore surfaces is the crucial determinant of the diffusion depth. An analytical model was developed to predict the gas molecular diffusion depth in the pore channels. Two different crystal sizes of UiO-66 were synthesized to validate against the model result and empirically determine the diffusion depths. The parametric effects of the mean free path, molecular kinetic energy, and molecular polarity on molecular diffusivity were assessed. A smaller molecular mean free path and greater molecular kinetic energy were favorable for larger diffusion depth, owing to the fewer collisions and enhanced bounces after collisions. Greater molecular polarity led to shorter diffusion depth due to the enhanced van der Waals force between molecules and pore surfaces. Full article

In the proposed centrifugal filtration device, a filter is mounted in the center of the centrifugal chamber. The particles move towards the centrifuge wall away from the filter under centrifugal force, so a filtration cake is unlikely to accumulate here. The working fluid is injected continuously, so the feed pressure is higher than the discharge pressure, which compels the light-phase fluid to leave through the filter as filtrate. The filtrate flux rate and the movement of particles in the centrifugal chamber of the proposed filtration device were investigated using different powder particle sizes, concentrations of working fluid, centrifugal chamber rotation speeds, and filters. With a higher centrifugal chamber rotation speed, the centrifugal force acting on the particles in the centrifugal chamber was stronger, and the particles were less likely to adhere to the central filter; hence, a larger amount of filtrate was produced. Full article

This study aims to screen and characterize the protective effect of polysaccharides from Portulaca oleracea L. (POP) against H₂O₂-stimulated osteoblast apoptosis in vivo and in vitro. The enzymes viscozyme, celluclast, α-amylase, and β-glucanase were used to extract POPs. Among all enzyme-assisted POPs, the first participating fraction of viscozyme extract POP (VPOP1) exhibited the highest antioxidant activity. Hoechst 33342 and acridine orange/ethidium bromide staining and flow cytometry of MC3T3 cells revealed that VPOP1 inhibited apoptosis in a dose-dependent manner. Moreover, VPOP1 increased the expression levels of heme oxygenase-1 (HO-1) and NADPH quinine oxidoreductase 1 (NQO1) and decreased the expression levels of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) in H₂O₂-induced cells compared with their controls. The results of an in vivo experiment show that VPOP1 significantly reduced reactive oxygen species generation and lipid peroxidation in zebrafish at 72 h post-fertilization and promoted bone growth at 9 days post-fertilization. Furthermore, VPOP1 was identified via 1-phenyl-3-methyl-5-pyrazolone derivatization as an acidic heteropolysaccharide comprising mannose and possessing a molecular weight of approximately 7.6 kDa. Collectively, VPOP1 was selected as a potential anti-osteoporotic functional food because of its protective activity against H₂O₂-induced damage in vitro and in vivo. Full article

The methanolic extract from the flowers of Mesua ferrea Linn. (Calophyllaceae) showed significant hyaluronidase inhibitory activity. Following a bioassay-guided separation of the extract, two biflavonoids, viz., mesuaferrone-A (1) and mesuaferrone-B (2), were isolated, along with ten flavonoids (3–12), two xanthones (13 and 14), three triterpenes (15–17), a phenylpropanoid (18), and five aromatics (19–24). Among the isolates, 1 and 2 (IC₅₀ = 51.1 µM and 54.7 µM, respectively) exhibited hyaluronidase inhibitory activity equivalent to that of the commercially available antiallergic agents disodium cromoglycate (64.8 μM) and ketotifen fumarate (76.5 μM). These biflavonoids (1 and 2) are 8-8″ linked dimers that are composed of naringenin (1a) or apigenin (3), with their corresponding monomers lacking inhibitory activity (IC₅₀ > 300 μM). In addition, 1 and 2 (IC₅₀ = 49.4 µM and 49.2 µM, respectively) inhibited the release of β-hexosaminidase, which is a marker of antigen-IgE-mediated degranulation, in rat basophilic leukemia (RBL-2H3) cells. These inhibitory activities were more potent than those of the antiallergic agents tranilast and ketotifen fumarate (IC₅₀ = 282 μM and 158 μM, respectively), as well as one of the corresponding monomers (1a; IC₅₀ > 100 μM). Nonetheless, these effects were weaker than those of the other monomer (3; IC₅₀ = 6.1 μM). Full article

The aim of the project was to investigate Peat-derived Fulvic acid for its propensity to form co-crystals with quercetin and curcumin and characterize it by using different analytical techniques. The formation of co-crystals generally enhances water solubility and the overall bioavailability of molecules. Co-crystals were synthesized using a 1:1 stoichiometric ratio of fulvic acid with quercetin and curcumin, respectively, using solvent crystallization techniques taking tetrahydrofuran and water in a 1:1 v/v ratio. The co-crystals were characterized by spectroscopic methods, FTIR and Differential scanning calorimetry. Further confirmation was made by morphological studies using SEM. A structural analysis was also carried out, using ¹³C solid-state NMR analysis. The studies confirmed the formation of semi crystalline forms. Furthermore, the saturation solubility displayed the enhancement in solubility of up to 10, 5-folds for Quercetin and Curcumin, respectively. The in vitro dissolution results showed that T50% was achieved within 30 min for both the drugs. The literature supports that the nutraceutical co-crystals offer advantages, particularly in the improvement of biopharmaceutical properties and addressing the challenges of the lab and manufacturing scale process. Both the semi crystalline powders exhibited enhanced solubility and a better dissolution profile. Full article

Peptides are a class of biomolecules with a great potential from the therapeutic point of view, because of their unique biological properties. Industrially, the production stategies adopted produce both the target peptide and a series of impurities that must be removed. Preparative chromatography is the technique of choice for the large-scale purification of biomolecules, generally performed in reversed-phase mode, using hydrophobic adsorbents (e.g., C8 stationary phases). A promising and innovative alternative is represented by mixed-mode columns, which bear two different ligands on the particle surface, exploiting two different retention mechanisms to improve the separation. This work represents a proof-of-concept study focused on the comparison of a hydrophobic adsorbent and a mixed-mode one (bearing both hydrophobic groups and charged ones) for the purification of a crude peptide mixture. Thanks to more-favourable thermodynamics, it was found that, when collecting the whole peak excluding fractions of the peak tail, the mixed-mode column led to an increase in the recovery of roughly +15%, together with a slight improvement in purity at the same time, with respect to the traditional hydrophobic column. In addition, if the whole peak, including the tail, is collected, the performance of the two columns are similar in terms of purity and recovery, but the pepetide elutes as a narrower peak with the mixed mode. This leads to a collection pool showing a much-higher peptide concentration and to lower solvent volumes needed, which is a beneficial achievement when targeting more sustainable processes. These results are very advantageous from the industrial viewpoint, because they also involve a decrease in the peptide amount contained in the peak tail, which must be reprocessed again to satisfy purity requirements. Full article

Short-chain fatty acids (SCFAs), produced by microbes when dietary fiber ferments in the colon, are one of the most studied microbial products despite their volatility and complex matrices, which make analysis challenging. In the current study, we sought to address research gaps by exploring the commonalities and differences between the retention time changes for SCFAs in polar solvents. In one such solvent, dimethyl sulfoxide (DMSO), the retention time of the SCFA acetic acid shows a linear positive correlation with the equal volume increase in the DMSO solvent. We used gas chromatography–mass spectrometry to analyze the retention times of mixed solutions of formic acid, acetic acid, butyric acid, valeric acid, and toluene in the solvents DMSO and water and found that only the retention times of formic acid and acetic acid changed. We further compared the effect of three solvents with similar polarities, DMSO, N-methylpyrrolidone (NMP), and dimethylformamide (DMF), on the retention time of acetic acid and found that it increased in the DMSO–water solution more than in the NMP–water solution and remained unchanged in the DMF–water solution. This finding is consistent with quantum chemical calculations showing that the strength of the hydrogen bond between DMSO and acetic acid is greater than between NMP and acetic acid. Taken together, the chromatographic results and quantum chemical calculations indicate that, in all three solvents, the portion of the molecule with the smallest negative electrostatic potential (red) has high electron density and can easily donate electrons, forming a hydrogen bond with acetic acid. However, the portion with the largest positive electrostatic potential (blue) forms a bond with polyethylene glycol, a column stationary solution with a strong dipole moment, and is adsorbed on the stationary solution in the direction of the dipole moment. Therefore, the retention times of formic acid and acetic acid change under the combined influence of a series of complex intermolecular forces. In the chromatographic column, the outflow rate of DMF is higher than that of acetic acid, and the force of the hydrogen bond between DMF and acetic acid cannot overcome the outflow resistance of acetic acid, so the retention time of the acetic acid in the DMF–water solution does not change. The retention times of butyric acid and valeric acid are unchanged in aprotic polar solvents for the same reason. Full article

This paper illustrates the origin, mission, organization and main activities of the Interdivisional Group of Separation Science (Gruppo Interdivisionale di Scienza delle Separazioni, GISS) of the Italian Chemical Society in occasion of the twentieth anniversary of this organization, whose mission is to promote and disseminate knowledge on all aspects of separation science and to facilitate strong peer-to-peer relationships. GISS originates from a discussion group, also belonging to the Italian Chemical Society (SCI), having similar finality and the same promoter (the Italian pioneer of Chromatography Prof. Arnaldo Liberti), but less extended scientific interests than the GISS, since it was mainly focused on liquid and gas chromatography. The interests of the GISS span theoretical and practical aspects of separation science and cover all key topics requiring high-performance separation techniques to carry out advanced investigations and/or solve challenging analytical problems. The panoramic view of the GISS includes a synthetic review of the main activities carried out since its foundation, which comprises fast-track discussion meetings, participation, promotion and/or organization of national and international congresses, sponsorship of participation grants for young trainees and Ph.D. students, recognition of scientists who have made and continue to make an outstanding contribution to separation science, as well as young scientists who have evidenced their attitude to carry out significant studies in the field of separation science. Full article

Computational fluid dynamics (CFDs) were adopted in order to investigate the solid suspending process in a dense solid–liquid system (with a solid volume fraction of 30%), agitated by a traditional dual axial impeller and a modified dual axial impeller, otherwise known as a dual triple blade impeller (DTBI) and a dual rigid-flexible triple blade impeller (DRFTBI), respectively. The effects of rotational speed, connection strap length/width, and off-bottom clearance on the solid distribution were investigated. The results show that the proportion of solid concentration larger than 0.4 in the DTBI system was 26.56 times of that in the DRFTBI system. This indicates that the DRFTBI system can strengthen the solid suspension and decrease the solid accumulation in the bottom of the tank. Furthermore, the velocity and turbulent kinetic energy in the DRFTBI system were promoted. In addition, for an optimal selection, the optimum length of connection strap was 1.2 H₁, the optimum range of connection strap width was D/7–D/8, and the off-bottom clearance selected as T/4 was better. Full article

A rapid and simplified sample preparation method was developed for the simultaneous determination of 26 beta-agonists in swine muscle using a multi-functional filter (MFF) based on quick, easy, cheap, effective, rugged, and safe methods (QuEChERS). MFF integrated the cleanup and filter procedures, thereby significantly improving the efficiency of sample preparation compared with traditional solid-phase extraction. The sample was processed via enzymatic hydrolysis, purified with the optimized MFF containing 150 mg magnesium sulfate, 50 mg PSA, and 50 mg C18, and then analyzed using ultra-high performance liquid chromatography-tandem mass spectrometry. All procedures can be completed in 6.5 h. Good linearity (R² > 0.99) was detected in all analytes. The recoveries ranged from 71.2% to 118.6%, with relative standard deviations (RSDs) of less than 18.37% in all spiked concentrations. The limits of detection (LOD) and the limits of quantitation (LOQ) were 0.01–0.10 and 0.10–0.50 μg/kg, respectively. The decision limit (CCα) and detection capacity (CCβ) values fluctuated in the range of 3.44–25.71 and 6.38–51.21 μg/kg, respectively. This method is a good alternative for detecting beta-agonist residues in swine muscle and can be successfully applied to the national risk monitoring of agro-product quality and safety in China. Full article

Microalgae are now known as potential microorganisms in the production of chemicals, fuel, and food. Since microalgae live in the sea and the river, they need to be harvested and separated and cultured for further usage. In this study, to separate microalgae, a bed of magnetite-containing gel (Mag gel, 190 µm) was packed in the column by the application of a magnetic field for the separative elution of injected microalgae (including mainly four species), cultured at Saga University in Japan. The applied magnetic field was set at a constant and dynamic-convex manner. At a constant magnetic field of 0.4–1.1 T, the elution percentage of the microalgae at less than 5 µm was 30–50%. At 1.1 T, the larger-sized microalgae were eluted at a percentage of 20%, resulting in the structural change of the bed by the applied magnetic field. In a convex-like change of the magnetic field at 1.1 T ⇄ 0.4 T, the smaller-sized microalgae were selectively eluted, whereas at 1.1 T ⇄ 0.8 T, the larger-sized microalgae were eluted. Dynamic convex-like changes by the magnetic field selectively eluted the microalgae, leading to the separation and the extraction of potential microalgae. Full article

In this work, the stability of antioxidant compounds in malting barley seeds before and after the production of the final products is reported. In this reflection, the findings revealed that the process of fermentation had a significant impact on antioxidant activity. In vitro, antioxidant capacities were evaluated using DPPH free radical scavenging assay. The results obtained from the spectrophotometric analysis showed that the lowest inhibition value was observed in the samples that were obtained by the classical fermentation process (ABC) and the samples of non-alcoholic beer obtained by the thermal process (NABT), with free radical inhibition capacity values of 8.50% and 5.50%, respectively. The samples of hopped wort (HW) and malted barley seeds extract (BSE) showed very high antioxidant activity with free radical inhibition capacity of 14% and 12.60%, respectively. The obtained extracts were analyzed by gas chromatography and high-performance liquid chromatography, both combined with mass spectrometry detection (GC–MS, HPLC–MS). GC–MS analysis of the SPME extraction showed the presence of 29 compounds with isopentyl alcohol in major concentration (18.19%) in the alcoholic beer; on the other hand, the HPLC–DAD–ESI/MS analysis of the ethyl acetate extract showed the presence of 13 phenolic compounds. Interestingly, the degradation of 3-Hydroxyphloretin 2′-O-glucoside in the final products of the non-alcoholic beers was found. Finally, the FTIR analysis was also employed in order to detect the type of efficient groups present in the extracts. Full article

A simple analytical approach based on capillary zone electrophoresis with ultraviolet detection and repeated sample injection strategy (applied in a hydrodynamically closed separation system for the first time) was developed for the determination of ibuprofen (IBU) in commercially available pharmaceutical preparations. The proposed method was characterized by significantly increased sample throughput and favorable validation parameters, highly demanded in routine quality control laboratories. The limit of detection was predicted at the concentration level of 0.31 µg/mL. Intra-day precision expressed as the relative standard deviation of IBU concentration ranged from 1.9 to 5.6%, and corresponding intra-day accuracy expressed as the relative error was in the interval of 87.1–106.5%. Inter-day precision was in the range of 2.6–15.0%, and inter-day accuracy was 94.9–102.7%. The developed method was able to quantify IBU in complex pharmaceutical matrices represented by commercially available tablets and oral suspension. The determined contents of IBU in the tested dosage forms were in good agreement with the manufacturer’s declaration. The analytical performance of the developed method was evaluated according to the innovative RGB Additive Color Model strategy. It was demonstrated that the proposed method is characterized by very good analytical performance parameters, safety and eco-friendliness, and practical effectiveness. Full article

The main objective discussed in this research was to determine the structural characteristics of carob kibble water-soluble polysaccharide (KWSP), extracted from Ceratonia siliqua L., and to assess its in vitro antioxidant activities, as well as its effect on whipped cream. The results obtained through ¹³C, ¹H, and the hetero-nuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FT-IR) analysis indicated that KWSP had the characteristic bands of polysaccharides. Thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) suggested that KWSP is a hetero-polysaccharide composed of glucose and fructose, with an average molecular weight (Mw) amounting to 65 KDa. In addition, KWSP showed a good water holding capacity (WHC), a good oil holding capacity (OHC), and an emulsification stability, rated as 3.14 ± 0.05 g/g, 0.87 ± 0.02 g/g, and 71 ± 0.01%, respectively. The antioxidant activity of KWSP was investigated in vitro, demonstrating important DPPH and ABTS⋅+ radical scavenging activities and a good total antioxidant capacity. KWSP exhibited antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Salmonella enterica. On the other hand, the incorporation of KWSP in whipped cream was investigated, to enhance its antioxidant capacity and consequently to extend its expiration date. Moreover, KWSP reduces the formation of conjugated dienes and trienes in cream fat. Full article

Volatile organic compounds (VOCs) are important sources of atmospheric pollutants on account of their high recycling value. The membrane of dense silicone rubber polydimethylsiloxane (PDMS) has wide-ranging prospects for the separation and recovery of VOCs. In this study, PDMS membrane body models were established in BIOVIA Materials Studio (MS) to simulate VOCs with C₃/N₂ gases, and to study the structure of PDMS membranes and the dissolution and diffusion process of gas in the membranes. The free volume fraction (FFV), cohesive energy density (CED), radial distribution function (RDF), diffusion coefficient and solubility coefficient of C₃H₈, C₃H₆ and N₂ in PDMS membranes were calculated, and the permeability coefficients were calculated according to these values. At the same time, the effects of temperature and mixed gas on the dissolution and diffusion of C₃/N₂ in PDMS membranes were investigated. The results show that the mass transfer process of C₃ in PDMS membranes is mainly controlled by the dissolution process, while that of N₂ is mainly controlled by the diffusion process. In a C₃/N₂ mixed gas system, there is a synergistic relationship between gases in the diffusion process, while there is competitive adsorption in the dissolution process. With an increase in temperature, the diffusion coefficients of the three gases in PDMS gradually increase, the solubility coefficients gradually decrease, and the overall permeability selectivity coefficients of the gases gradually decrease. Therefore, low-temperature conditions are more conducive to the separation of C₃/N₂ in PDMS membranes. The simulation results of the permeability selectivity coefficients of pure C₃ and N₂ in PDMS are similar to the experimental results, and the relationship between the micro- and macro-transport properties of PDMS membranes can be better understood through molecular simulation. Full article

Calycosin is a dietary flavonoid with favorable activities, which seems to be inconsistent with its low circulating levels in vivo. To address this issue, we developed a strategy to understand calycosin distribution by integrating qualitative and quantitative analyses of calycosin and its metabolites in portal vein plasma, the liver, and systemic plasma after oral administration to rats. Consequently, 21 metabolites were characterized in total, including the first report of a reductive biotransformation and 14 new metabolites. Compared with the low levels of calycosin, calycosin glucuronides were predominant in circulation, and both the hepatic and intestinal regions contributed to the high exposure of these calycosin glucuronides. However, intestinal glucuronidation, prior to hepatic glucuronidation, plays a key role in the low circulating levels of calycosin. Full article

The development of chromatographic science in Ukraine is due to the widespread use of chromatography to solve practical problems. The centers of chromatographic research in Ukraine are in large cities: Kyiv, Kharkiv, Lviv, Odessa, and others. Along with the development of chromatographic research methods, analysis methods for control of food and agricultural raw materials, medicinal products, petroleum products, and determination of the environmental pollution state are created. Chromatography is used in medicine, pharmacology, biology, and other sciences. Full article

Ribavirin, a nucleoside analog, is used to treat chronic hepatitis C (HCV) infections. Therapeutic drug monitoring for ribavirin is useful for predicting the effect of treatment. In this study, the selective extraction of ribavirin from serum samples and the HPLC-UV detection method were investigated using a monolithic silica disk-packed spin column with phenylboronate moieties. In this study, 0.6% ammonia and 1% formic acid solutions were used as the conditioning and elution solutions, respectively, and recoveries of >90% were obtained. Ribavirin was separated on an InertSustain AQ-C18 column by isocratic elution. The mobile phase consisted of a mixture of 7 mM Na₂SO₄ and 60 mM H₃PO₄ in H₂O. Linear regression curves were observed for calibrations over a concentration range of 0.25–25 µg/mL. The lower limit of detection was 0.05 µg/mL, and the lower limit of quantification was 0.1 µg/mL. The intra- and inter-day precisions were below 3.2 and 3.1%, respectively. This method can be applied to quantify ribavirin levels in human serum and may be useful for pharmacokinetic studies. Full article

Formaldehyde (FA) is a toxic compound that is considered to have a carcinogenic effect due to its damage to biological macromolecules. However, the influence of FA at the protein level remains to be explored. Here, we used LC-MS/MS to identify the differentially expressed proteins and modifications to proteins between FA-treated and untreated HeLa cells. Among 2021 proteins identified, 196 proteins were significantly down-regulated and 152 up-regulated. The differentially expressed proteins were further analyzed using bioinformatics tools for annotating the characterization of their localizations and functions. To evaluate the interaction of FA with proteins, we performed proteomic analysis for a mass shift of 12 Da on the side chains of lysine, cysteine and tryptophan, which are induced by FA as noticeable signals. We identified the modified proteins and sites, suggesting direct interaction between FA and proteins. Motif analysis further showed the characterization of amino acid sequences that react with FA. Cluster analysis of the modified proteins indicated that the FA-interacting networks are mostly enriched in the nuclei, ribosomes and metabolism. Our study presents the influence of FA on proteomes and modifications, offering a new insight into the mechanisms underlying FA-induced biological effects. Full article

The health benefits of Dracocephalum heterophyllum are widely reported in traditional Tibetan medicines, but the reported chemical composition is limited, probably due to difficulties in separating and purifying compounds. In this study, antioxidative phenylethanoids were isolated from an extract of Dracocephalum heterophyllum using medium- and high-pressure liquid chromatography, coupled with on-line HPLC–1,1-diphenyl-2-picrylhydrazyl recognition. Firstly, crude samples (1.3 kg) of Dracocephalum heterophyllum were pretreated via silica gel medium-pressure liquid chromatography to yield 994.0 g of Fr2, of which 10.8 g was then pretreated via MCI GEL^®CHP20P medium-pressure liquid chromatography. The resulting Fr23 and Fr25 were further separated and purified using high-pressure liquid chromatography, and yielded 8.08 mg of Fr2391, 9.76 mg of Fr2551, 16.09 mg of Fr2581, and 8.75 mg of Fr2582. Furthermore, analysis of the purity and structures of the phenylethanoids suggested that Fr2391, Fr2551, Fr2581, and Fr2582 corresponded to decaffeoylverbascoside, rosmarinic acid, acteoside, and 2′-O-acetylplantamajoside, respectively, with all being over 95% pure. Finally, the antioxidant potential of the compounds was explored based on their ability to scavenge 1,1-diphenyl-2-picrylhydrazine, as well as through molecular docking of proteins related to antioxidant pathways. Altogether, our findings revealed that the proposed method is promising for separating pure antioxidative phenylethanoids from other natural compounds. Full article

The most common beneficiation method for feldspar is flotation with a cationic (amine) collector under acidic conditions. However, there are several disadvantages to this, such as environmental pollution and equipment corrosion. In order to resolve such problems, it is important to study the flotation of feldspar using anionic collectors under natural pH conditions. The purpose of this paper is to study the effects and mechanism of Fe³⁺ on flotation separation of feldspar and epidote using sodium oleate (NaOL) at a natural pH. Through flotation experiments, adsorption measurements, zeta potential testing, FTIR analysis and X-ray photoelectron spectroscopy (XPS), the mechanism of Fe³⁺ on the surface of feldspar and epidote is revealed, and the reason behind the difference in flotation of the two minerals is discussed. The flotation test results show that Fe³⁺ can significantly improve the flotation behavior of minerals when NaOL is used as a collector under natural pH, and the highest recovery rates of feldspar and epidote are 90% and 43%, respectively. Analysis of the solution and adsorption measurement results show that Fe³⁺ is adsorbed on the minerals′ surface in the form of Fe(OH)₃, which promotes the adsorption of NaOL on the minerals’ surface through Fe(OH)₃, activating the flotation of feldspar and epidote. The difference in adsorption of Fe³⁺ between feldspar and epidote is the reason for this difference in flotation behavior. The results of the zeta potentials show that after being treated with Fe³⁺, the electrostatic adsorption of NaOL displays a significant negative shift on the surface of feldspar, while there is almost no electrostatic adsorption of NaOL on the surface of Fe³⁺-treated epidote. FTIR analysis confirmed that the difference in the adsorption of Fe³⁺ and NaOL on the surface of feldspar and epidote is due to the fact that there are more active particles (metal bonds) on the surface of feldspar than on the surface of epidote, and the properties of these metal bonds can be changed by Fe³⁺, which allows NaOL to be more easily adsorbed on the mineral surface through –COO⁻. The possible adsorption form is “mineral-Fe³⁺–COO⁻“. Compared with the infrared spectrum of epidote, there is a new absorption peak at 1713.68 cm⁻¹, which can be attributed to the C=O characteristic peak of NaOL in the infrared spectrum of Fe³⁺–NaOL-treated feldspar, which is why the floatability of feldspar is better than epidote. XPS confirmed that the Fe on the surface of feldspar is Fe³⁺ in the form of Fe(OH)3, while Fe on the surface of epidote is mainly Fe₂O₃ (Fe–O) contained in mineral crystals. Furthermore, there is less adsorption of Fe³⁺ on the surface of epidote, and this discrepancy leads to the difference in the adsorption of NaOL on the minerals’ surface, which itself leads to the difference in flotation behavior between feldspar and epidote. These findings indicate that the flotation separation of feldspar and epidote can be achieved using Fe³⁺ and NaOL under natural pH. This study may provide a reference for the flotation mechanism of feldspar and epidote under natural pH. Full article

In this study, in order to protect the characteristic Dendrobium devonianum Paxt industry in the Longling area, and promote the healthy development of its characteristic Chinese herbal medicines in Yunnan Province, China, the identification of Dendrobium devonianum Paxt and Dendrobium officinale Kimura et Migo from Longling county was discussed using time of flight mass spectrometry. The data of 13 Dendrobium devonianum and 7 Dendrobium officinale in the Longling area were collected by TOF MS-IDA-15 MS/MS mode, and the collected data were analyzed by PCA and T-test using MarkerView software, and the difference markers were searched using the database to confirm their compound structures. In positive and negative ion modes, 3645 and 2344 peaks were detected, respectively; 64 positive ion compounds and 60 negative ion compounds, for a total of 124 compounds were identified, mainly including organic acids, polyphenols, alkaloids, amino acids and their derivatives, benzene and its derivatives, and other compounds. The score plot and loading plot analyzed by PCA show that Dendrobium devonianum and Dendrobium officinale collected in the Longling area can be effectively identified and differentiated by high-resolution mass spectrometry with the 15 different markers in positive ion mode and 17 markers in negative ion mode, respectively. The successful identification of Dendrobium devonianum and Dendrobium officinale fully demonstrates that TOF MS can be effectively used in the identification of Dendrobium and related Chinese herbal medicines with broadly application foreground. Full article

The demand for skin-whitening agents is high across the world, including Asian countries. An extensive screening using a tyrosinase inhibition assay was performed in order to discover novel plant materials. In our research program investigating a safe and effective agent, 50% ethanolic extracts prepared from discarded parts of Prunus persica were screened for in vitro tyrosinase inhibitory activity. Among the extracts tested, twig extract showed the most potent inhibitory activity: 38% inhibition at 500 µg/mL. The investigation of active compounds in twig extract found four flavanones that acted as moderate inhibitors, including (−)-prunin, persiconin, (+)-dihydrokaempferol, and (−)-naringenin. These compounds were only observed in the twig extract following preliminary quantification by HPLC, with the following concentrations: (−)-prunin, 1.8 mg/g sample; persiconin, 0.8 mg/g sample; (+)-dihydrokaempferol, 0.8 mg/g sample; (−)-naringenin, 1.7 mg/g sample. These results suggest that twig extracts can be more useful for skin-whitening compared with other parts of the plant. In addition, a new constituent of twig extract was identified, namely isoquercitrin, which suggests that twig extract can be a potent source of flavones and flavanones. Further studies on the identification of novel compounds from twig extract are now underway in our laboratory. Full article

The separation and discrete detection of isomeric sequence peptides with similar properties are important tasks for analytical science. Three different peptide isomers of 12 amino-acid residues long, containing direct and reverse regions of the alanine-valine-proline-isoleucine (AVPI) motif, were partially separated and discretely detected from their mixture using two approaches. Capillary electrophoresis enabled the separation and optical detection of the peptide sequence isomers close to the baseline. The ability to separate these sequence isomers from the mixture and discretely identify them from mass spectra has also been demonstrated by ion-mobility tandem mass spectrometry. Moreover, for the first time, capillary electrophoresis and ion-mobility mass spectrometry connected online have shown their ability for a discrete detection of the multidirectional sequence isomers. Full article

It is well established that arsenic (As) has many toxic compounds, and in particular, inorganic As (iAs) has been classified as a type-1 carcinogen. The measuring of As species in rice flour is of great importance since rice is a staple of the diet in many countries and a major contributor to As intake in the Asian diet. In this study, several solvents and techniques for the extraction of As species from rice flour samples prior to their analysis by HPLC-ICP-MS were investigated. The extraction methods were examined for their efficiency in extracting various arsenicals from a rice flour certified reference material, NMIJ-7532a, produced by the National Metrology Institute of Japan. Results show that ultrasound-assisted extraction at 60 °C for 1 h and then heating at 100 °C for 2.5 h in the oven using a thermostable α-amylase aqueous solution was highly effective in liberating the arsenic species. The recoveries of iAs and dimethylarsinic acid (DMA) in NMIJ-7532a were 99.7% ± 1.6% (n = 3) and 98.1% ± 2.3% (n = 3), respectively, in comparison with the certificated values. Thus, the proposed extraction method is a green procedure that does not use any acidic, basic, or organic solvents. Moreover, this extraction method could effectively maintain the integrity of the native arsenic species of As(III), As(V), monomethylarsonate (MMA), DMA, arsenobetaine (AsB) and arsenocholine (AsC). Under the optimum extraction, chromatography and ICP-MS conditions, the limits of detection (LOD) obtained were 0.47 ng g⁻¹, 1.67 ng g⁻¹ and 0.80 ng g⁻¹ for As(III), As(V) and DMA, respectively, while the limits of quantification (LOQ) achieved were 1.51 ng g⁻¹, 5.34 ng g⁻¹ and 2.57 ng g⁻¹ for As(III), As(V) and DMA, respectively. Subsequently, the proposed method was successfully applied to As speciation analysis for several rice flour samples collected from contaminated areas in China. Full article

A screening method using gas chromatography quadrupole time-of-flight mass spectrometry (GC-QTOF/MS) combined with mini solid-phase extraction (mini-SPE) was established for the quantification and validation of multiclass pesticide residues in tobacco. The method was quicker and easier, with sample purity higher than that obtained by traditional SPE and dispersed-SPE. Box-Behnken design, an experimental design for response-surface methodology, was used to optimize the variables affecting the target pesticide recovery. Under the optimized conditions, 92% of the pesticides showed satisfactory recoveries of 70%–120% with precision <20% at spiking levels of 50, 250, and 500 ng/g. The limits of detection and quantification for all the analyses were 0.05–29.9 ng/g and 0.20–98.8 ng/g, respectively. In addition, a screening method based on the retention time and a homebuilt high-resolution mass spectrometry database were established. Under the proposed screening parameters and at spiking levels of 50, 100, and 500 ng/g, 76.6%, 94.7%, and 99.0% multiclass pesticide residues were detected, respectively, using the workflow software. The validated method was successfully applied to the analysis of real tobacco samples. Thus, the combination of mini-SPE and GC-QTOF/MS serves as a suitable method for the quantitative analysis and rapid screening of multiclass pesticide residues in tobacco. Full article