Crassula species are traditionally used and possess anti-inflammatory properties, but Crassula tetragona L. remains largely unexplored. This study intended to characterize C. tetragona aerial parts' phytoconstituents and assess its anti-ulcerative potential via the PPARγ/SIRT1 pathway. Aerial parts of C. tetragona were extracted using n-hexane (CT1) and 70% aqueous methanol (CT2). Phytoconstituents were profiled by HPLC-ESI-MS/MS (negative ion mode), and phenolics were quantified by MRM-LC-ESI-MS/MS. Column chromatography and NMR were used to separate and identify the compounds. Ulcerative colitis (UC) was induced in rats by intrarectal acetic acid (AA). Animals were assigned into six groups: control group: orally received vehicle for 7days, UC control group: orally received vehicle for 7days, and a rectal infusion of 2mL AA (4% v/v in saline) on the 8th day, 4 treated groups: received CT1 (200 and 400mg/kg/day), or received CT2 (200 and 400mg/kg/day), once daily for 7days by oral gavage and 2mL AA (4% v/v in saline) on the 8th day. HPLC-ESI-MS/MS identified 66 constituents, including 37 novel compounds, with CT2 exhibiting higher phenolic content. Naringenin, gallic acid, and quercetin were predominant. Five phenolic compounds were isolated from the bioactive extract CT2. Both CT1 and CT2 reduced AA-induced tissue damage, lowered inflammatory markers (calprotectin, CRP, TNF-α, IL-6), improved oxidative stress (reduced MDA, increased GSH, SOD), and upregulated SIRT1 and PPARγ. These results suggest C. tetragona attenuates UC via the SIRT1/PPARγ pathway, indicating its therapeutic potential.

Bioactivity-guided isolation of pancreatic lipase inhibitors from Lycium ruthenicum using a novel HPLC-FLD/pancreatic lipase online recognition system.

Eutectogel-mediated metal organic framework@silica composites: Enabling multi-mode separation in high-performance liquid chromatography.

Pyrrolizidine alkaloids （PAs） are natural toxins widely distributed in plants， which naturally occur in about 3% of the world's flowering plants. To date， more than 660 PAs and their nitrogen oxides have been identified in over 6 000 plants. Some PAs are hepatotoxic， genotoxic and tumorigenic， posing significant health risks to humans. These alkaloids are commonly detected as contaminants in various food products， including tea， grains， milk， honey， as well as plant-derived pharmaceuticals and dietary supplements. Currently， most studies on the quantitative methods for PAs focus on a limited number of PAs and employ an additive quantification strategy， largely due to the challenges associated with chromatographic separation of isomers. These approaches limit the ability to assess exposure and health risk accurately. Herein， a method was established to quantify 35 PAs individually in dried tea samples using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry （UHPLC-MS/MS）. The 35 target compounds were divided into two groups. The first group included 30 PAs， while the second group consisted of 5 PAs. These compounds were separated on Waters Acquity BEH C18 （150 mm×2.1 mm， 1.7 μm） and Thermo AcclaimTM C30 （150 mm×2.1 mm， 3.0 μm） chromatographic columns， respectively. Mobile phases were H2O containning 5 mmol/L ammonium formate and 0.13% formic acid （pH=3） and methanol-acetonitrile （4∶6， volume raio） containing 0.1% formic acid. The 33 target compounds were separated and 2 isomers co-eluted. Under positive-ion electrospray ionization （ESI） and multiple reaction monitoring （MRM） mode， target compounds were quantified using the external standard method with the matrix-matched standard curve. The results demonstrated that all target compounds showed good linearity （r2>0.99） in their respective mass concentration ranges. The limits of detection （LODs） and quantification （LOQs） of the method were in the range of 0.2-8.0 µg/kg and 0.5-25.0 µg/kg， respectively. The average recoveries of more than 89% compounds were in the range of 70%-130% at spiked levels of 1， 2， and 5 fold-LOQs and the relative standard deviations （RSDs） were less than 20% （RSDs of lasiocarpine and echimidine were less than 30%）. Moreover， the quantitative method of the 35 PAs was applied to 21 dark tea and 30 black tea samples from Yunnan and Fujian Province， where PAs were detected in 4 dark samples with total contents ranging from 5.07 to 15.48 µg/kg and were not detected in black tea samples. The detection rate of PAs in dark tea samples was higher than that of black tea samples. That might be because most dark tea was processed from fresh leaves picked by machines， and this attribution could be associated with the mixing of weeds containing PAs during tea harvesting. And the detected concentrations of all samples were lower than the maximum levels of tea in European Union regulations， indicating that the tea samples involved in this research are basically safe. In brief， the quantitative method of the 35 PAs facilitates the analysis of the occurrence， composition and potential risks in tea samples.

As chemical agents that selectively inhibit weed growth， herbicides play a crucial role in enhancing crop yields. With increasing weed resistance， the environmental residue problems caused by excessive application have become increasingly prominent. Studies indicate that only 20%-30% of field-applied herbicides are effectively utilized， with the remainder entering environmental media such as the atmosphere， soil， sediment， and surface water through runoff and leaching. Recent frequent occurrences of vegetable phytotoxicity incidents in Shanghai have been traced to potential associations with herbicide residues in surface water， further highlighting the urgent need to establish multi-residue analytical methods for environmental media. An analytical method was established based on ultra-high performance liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS） for determining 26 herbicide residues in soil， sediment， and surface water. Instrumental detection parameters were optimized， including electrospray ionization mode， mobile phase， and chromatographic column. The mobile phase consisted of 0.1% formic acid aqueous solution （A） and acetonitrile （B） with the following gradient elution program： 0-0.5 min， 2%B； 0.5-1 min， 2%B-50%B； 1-4 min， 50%B-65%B； 4-6 min， 65%B-75%B； 6-8 min，75%B-85%B； 8-10 min， 85%B-95% B； 10-11 min， 95%B. Soil and sediment samples were extracted via acetonitrile oscillation followed by salting-out and purified using the QuEChERS method. Surface water samples were directly analyzed after acetonitrile extraction without purification. Different amounts of purification agents were investigated during sample pretreatment. Calibration curves were established by plotting the relationship between analyte concentration and measured peak area using pure solvent and matrix-matched standards. All 26 herbicides showed good linearity in the range of 0.1-50 μg/L with correlation coefficients exceeding 0.999 0. Matrix effects ranged from -35.2% to 14.6% across different matrices. Limits of quantification （LOQs） were 0.5 μg/kg for soil and sediment， 0.1 μg/L for water samples. The herbicides were spiked into soil and sediment at spiked levels of 0.5， 1， and 10 μg/kg， and into surface water at 0.1， 1， and 10 μg/L， respectively. The average recoveries for the 26 herbicides in soil， sediment， and surface water were in the ranges of 73%-108%， 73%-102%， and 74%-110%， respectively. The RSDs for the 26 herbicides were in the ranges of 4.5%-16.2%， 3.8%-19.7%， and 4.0%-15.0%， respectively. The developed method was applied to analyze the contamination status of the 26 herbicides in environmental samples collected from six vegetable cultivation zones in Shanghai. Results revealed distinct pollution patterns： In soil matrices， prometryn （PMT）， metolachlor （MTA）， and sulfometuron-methyl （SMTM） showed higher detection rates of 52.9%， 52.9%， and 29.4%， respectively， with content ranges of 0.8-490.4 μg/kg， 0.5-219.8 μg/kg， and 1.0-562.6 μg/kg. Sediment samples exhibited an 83.3% detection rate for PMT （1.5-6.7 μg/kg）. In surface water， SMTM， PMT， and simetryne （STN） were detected with maximum contents of 12， 2.5 and 1.1 μg/L， respectively， indicating differential migration behaviors across environmental compartments. The proposed method is simple， rapid， accurate， stable， and highly practical. It can be used to detect the 26 herbicide residues in soil， sediment， and surface water and provides a reference for monitoring the residual pollution and environmental behavior of herbicides.

Cadinane sesquiterpenes from Alangium platanifolium: structural elucidation and anti-colorectal cancer biological evaluation.

Candida species are the dominant inhabitants in the oral cavity and can potentially develop into opportunistic pathogens in immunocompromised individuals, leading to oral infections. The emergence of Candida resistance against antifungal drugs has called for an alternative. This study aimed to evaluate the antifungal efficacy of n-hexane extract and its bioactive compound, 1'-S-1'-acetoxychavicol acetate (ACA), isolated from the rhizome of Alpinia conchigera against oral Candida. A crude n-hexane extract (HE) was obtained from the dried, ground A. conchigera rhizome using the maceration technique. The HE was subjected to column chromatography to isolate ACA, followed by structural elucidation and purity confirmation using nuclear magnetic resonance (NMR) and high-performance liquid chromatography (HPLC). Oral Candida species were retrieved from the oral rinses of the older adults. The antifungal effects of HE and ACA against oral Candida isolates were screened using the disc diffusion assay, followed by an evaluation of minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC). A total of 21.391 g of HE was obtained, yielding 3.7573 mg of ACA with a purity of 93.67%. The largest inhibition zones were observed for HE and ACA, both at a concentration of 100 mg/mL against C. albicans (12.0 ± 0.8 mm) and C. glabrata (10.0 ± 0.8 mm), respectively. The MIC and MFC values of HE ranged from 0.7 to 12.5 mg/mL and from 0.7 to 25 mg/mL, respectively. Meanwhile, for ACA, both MIC and MFC values are within the range of 0.7 to 25 mg/mL. This in vitro study showed that both HE and ACA possess antifungal properties against Candida isolates, which highlights their potential as natural antifungal agents for incorporation into oral care products.

In the last few decades, many diseases have been caused by Radical Oxidative Stress (ROS) or free radicals. Compounds that act as natural antioxidants have been widely developed to counteract these free radicals. Rhizpora mucronata was a mangrove species reported to possess strong antioxidant activity. However, the research is limited to extracts; fractions with more specific compound components have not been explored. The n-hexane fraction of R. mucronata leaves was reported to have high total flavonoid content (TFC). The TFC has a positive correlation with antioxidant activity. Therefore, this study aimed to simplify the compound components in the n-hexane fraction of R. mucronata leaves using gravity column chromatography (GCG) and determine their antioxidant activity. The n-hexane fraction was fractionated using GCG with increasing solvent polarity (n-hexane, chloroform, ethyl acetate, and methanol). The fractions were tested for antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods. The fractionation using GCG resulted in 80 fractions. These fractions were grouped based on their thin-layer chromatography (TLC) profiles, resulting in 12 groups of fractions (A-L). Fractions D, F, G, H, J, K, and L, the fractions with enough yield, determined their antioxidant activity using the DPPH and ABTS methods. Fractions D and J tested by the DPPH method had IC50 values of 28.68 ± 1.58 ppm and 9.59 ± 0.39 ppm, respectively. Meanwhile, fractions D and J tested by the ABTS method had IC50 values of 15.10 ± 1.00 ppm and 3.16 ± 0.55 ppm, respectively. Meanwhile, fractions F, G, H, K, and L exhibit antioxidant activity with IC50 values greater than 100 ppm. Fractions D and J have potent antioxidant activity, and both were tested using DPPH and ABTS methods. Therefore, fractions D and J can be further developed as natural antioxidants.

Background: The global rise in visual impairment, driven by population aging, the increasing prevalence of lifestyle-related chronic diseases, and environmental factors, has made it a critical public health concern, highlighting the urgent need for effective preventive strategies and eye health maintenance. Cuscutae Semen (CS), a traditional Chinese herbal medicine long regarded for its vision-enhancing properties, has been widely used to support ocular health. However, its underlying molecular mechanisms and bioactive constituents remain poorly understood, limiting its modernization and broader clinical application. Objective: This study aims to investigate the restorative effects of CS on visual impairment, elucidate its underlying mechanisms, and identify potential active components. Methods: A zebrafish model of visual impairment was established using mepanipyrim to simulate retinal structural damage and visual dysfunction. The therapeutic effects of CS were systematically evaluated through behavioral analyses and histomorphological observations. To elucidate the underlying mechanisms, an integrated approach was employed, combining transcriptome sequencing (RNA-seq), reverse transcription quantitative polymerase chain reaction validation, and immunofluorescence staining to identify critical genes and pathways involved. Furthermore, macroporous resin column chromatography was employed for the fractionation and screening of potential active components. Results: CS treatment significantly alleviated mepanipyrim-induced ocular abnormalities in zebrafish, restoring approximately 82% of the observed morphological defects. Behavioral assessments revealed that CS-treated zebrafish exhibited markedly increased swimming speed and distance, indicating enhanced visual light sensitivity. Histopathological analysis demonstrated that CS effectively repaired the structure of retinal cell layers. RNA-seq revealed that CS broadly reversed mepanipyrim-induced gene expression disturbances, suggesting a restorative effect on transcriptomic homeostasis. Gene Ontology enrichment analysis identified the phototransduction pathway as a key mediator of CS’s therapeutic effects. This was further supported by reverse transcription quantitative polymerase chain reaction validation of critical genes and immunofluorescence staining, which confirmed the restored expression of Pde6a and Gnat2, key proteins involved in photic signal transmission. Active component screening indicated that high-polar constituents, including chlorogenic acid, may constitute one of the major bioactive fractions responsible for the observed therapeutic effects. Conclusion: This study provides evidence of the vision-protective effects of CS in a zebrafish model, demonstrating that its therapeutic mechanism involves modulation of the phototransduction pathway. Chlorogenic acid was identified as one of the key bioactive constituents contributing to this effect. These findings not only provide scientific validation for the traditional use of CS in ocular protection but also present promising therapeutic prospects for the prevention and treatment of visual impairment.

The mango (Mangifera indica L.) is a commonly cultivated tropical fruit across the globe. It is known to be rich in carotenoids, polyphenols, and vitamins, compounds that largely account for its nutritional and medicinal properties. Although the beneficial effects of mango phytochemicals have been widely documented, virtually no studies have investigated extracellular vesicles (EVs) originating from mango fruit. In the presented work, we developed a workflow combining differential centrifugation, filtration, and size-exclusion chromatography for the isolation of EVs from mango pulp. The isolates were characterized in accordance with the guidelines of the International Society of Extracellular Vesicles recommendations. The optimized size-exclusion chromatography column, packed with Sepharose CL-6B beads, enabled the recovery of a high-quality EV fraction, which was characterized in terms of physicochemical properties. Additionally, proteomic analysis identified 1084 proteins, many of which are associated with antioxidant, antimicrobial, and anti-inflammatory functions. These findings provide the first comprehensive characterization of mango-derived EVs and suggest that they may contribute to the biological activity traditionally attributed to mango consumption.

The lanthanide elements have a wide range of applications including electronics, radiopharmaceuticals, and nuclear energy materials. The isolation and separation of the lanthanides is challenging due to their similar chemistry, caused by their ubiquitous 3+ oxidation state and small differences in ionic radii across the series. Promethium (Pm) is the only lanthanide with no stable isotopes, resulting in the exclusion of Pm from most lanthanide studies, yet Pm is an isotope in demand by the US.1 To establish a national supply of Pm, the separation of neodymium (Nd), Pm, and samarium (Sm) is required. Oak Ridge National Laboratory (ORNL) has recently invested significant efforts in harvesting the fission byproduct Pm-147 from their plutonium-238 campaign.2 ORNL is the sole United States’ producer of Pm, so a tailored separation scheme for Pm applicable to the plutonium-238 campaign is essential. Current lanthanide separation schemes use their slight ionic size differences in the trivalent state; however, this results in low separation factors and extensive separation times.2,3 A novel method to facilitate the separation of neighboring lanthanides includes selectively reducing one lanthanide to allow for a Ln2+ Ln3+ separation. For this separation scheme to be realized, their differing electrochemical properties need to be investigated. The reduction potentials of Nd, Pm, and Sm from the 3+ to 2+ state have been experimentally determined to be -2.62 V, -2.44 V, and -1.50 V, respectively.4 Before applying our study to radioactive Pm, we will optimize with nonradioactive analogs, Nd and Sm. Nd is an acceptable analog for Pm due to its similar reduction potential, ionic radius, and available oxidation states. 18-crown-6 derivatives such as dicyclohexano-18-crown-6 (DCH18C6) have been shown to selectively coordinate and stabilize divalent lanthanides.5 Electrochemical measurements of DCH18C6 coordinated to Nd, Pm, and Sm will elucidate the stabilizing effect of this crown ether molecule. The positive shift in reduction potential for Sm and Nd will be determined through cyclic voltametric experiments. These methods can be further optimized by investigating the effect of different crown ether derivatives, various counter anions, and solvents on the reduction potential of Sm and Nd. For ultimate separation methods, coordination with a bulky nonpolar ligand can facilitate highly effective liquid-liquid extractions or column chromatography separations. In the future, determination of the best parameters to stabilize divalent Sm and Nd will be applied to the separation of Nd from Pm and Sm from Pm.(1) Compelling Research Opportunities Using Isotopes; DOESC (USDOE Office of Science (SC)), 2009.(2) Mayes, R. T.; VanCleve ,Shelley M.; Kehn ,Jay S.; Delashmitt ,Jordan; Langley ,Josh T.; Lester ,Brian P.; Du ,Miting; Felker ,L. Kevin; and Delmau, L. H. Combination of DGA and LN Columns: A Versatile Option for Isotope Production and Purification at Oak Ridge National Laboratory. Solvent Extr. Ion Exch. 2021, 39 (2), 166–183.(3) Ward, J.; Bucher, B.; Carney, K.; Snow, M. Exploring Lanthanide Separations Using Eichrom’s Ln Resin and Low-Pressure Liquid Chromatography. J. Radioanal. Nucl. Chem. 2021, 327 (1), 307–316.(4) Mikheev, N. B. Lower Oxidation States of Lanthanides and Actinides. Inorganica Chim. Acta 1984, 94 (5), 241–248.(5) Poe, T. N.; Molinari, S. E.; Wineinger, H. B.; Albrecht, T. E. Isolation of Inner-Sphere Aquo Complexes of Samarium(II). J. Am. Chem. Soc. 2025, 147 (3), 2323–2334.

Extracellular vesicles (EVs) are considered to be a promising tool in disease diagnosis. However, the clinical translation of EV-based liquid biopsy faces significant challenges due to the lack of inexpensive, rapid, and high-throughput methods of EV analysis. Bead-based platforms, combined with conventional flow cytometry, allow for the simultaneous capture and immunolabeling of EVs. In this study, we present a new approach based on the label-free isolation of EVs by tannic acid-coated superparamagnetic beads (TASPMB) combined with immunofluorescence detection of EV membrane proteins using flow cytometry. First, we tested the molecular profiling capabilities of the approach using EVs derived from human breast and colorectal cancer cell lines and from plasma of colorectal cancer patients to recognize the tetraspanin protein CD63 and the epithelial cell adhesion molecule (EpCAM). Subsequently, the developed approach was validated to identify proteins on EVs enriched with TASPMB from the conditioned media of SKBR3 and HT29 cell cultures without preliminary purification by a size-exclusion chromatography (SEC) column. The developed approach demonstrates a high capacity for isolating EVs and subsequently profiling of their membrane proteins, with a total assay time of approximately 2 h. The approach presented here can be a promising tool for rapid detection of EV membrane proteins using conventional instruments, such as flow cytometry.

Gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) is the method of choice to detect the abuse of synthetic forms of naturally occurring steroids for doping control purposes. GC-C-IRMS relies on a multistep sample cleanup to ensure each target analyte peak is chromatographically pure before combustion. To achieve that, liquid-liquid or solid phase extraction (SPE) is commonly used in combination with preparative liquid chromatography (LC). In this work, a procedure for isolation, purification, and analysis of endogenous steroids by GC-C-IRMS was developed and validated. The key elements were successive application of strong cation and strong anion exchange SPE with enzymatic hydrolysis in between to strip the ionic species from urine and decrease matrix complexity prior to LC cleanup; preparative two-dimensional LC, where only the testosterone fraction required secondary purification (40 min total run time per sample); and derivatization of selected fractions with formic acid to yield formate esters, followed by GC-C-IRMS analysis. Formylation afforded excellent separation between 5α- and 5β-androstanediols and simplified the detection of the endogenous reference compound pregnanetriol by converting it to a volatile artifact, tentatively identified as 3α,20-diformoxy-17-methyl-18-nor-5β,17α-pregn-13-ene. The overall method performance benefited from the customization of the GC-C-IRMS combustion interface, which improved robustness and facilitated the transfer of sample components into the hot zone of the oxidation reactor, minimizing peak tailing. The former was achieved by keeping the oxygen flow through the reactor at all times, obviating the need for periodic oxidation, and the latter-by implementing a direct capillary-in-capillary connection of the chromatographic column to the oxidation reactor.

We have developed a new hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-MS) method with mobile phases optimized for high metabolite coverage in individual polarity modes. This dual mobile phase strategy expands the range of annotated metabolites and improves identification confidence, providing broader and more accurate metabolic profiles. The incorporation of a bioinert chromatographic system further enhances sensitivity. The bridged ethyl hybrid (BEH) amide column yields the best results for metabolomic analysis among the three chromatographic columns in this comparison. The method development involves investigating the effects of mobile phase composition, pH, and a medronic acid additive on the MS response under bioinert chromatographic conditions. The results highlight the important role of alkaline pH for the sensitive detection of polyphosphorylated metabolites, while demonstrating the redundancy of chelating additives in a fully bioinert system. Finally, the optimized method is applied to mouse plasma, pancreas, and liver samples to demonstrate its versatility and reliable performance in complex biological matrices, establishing it as a powerful tool for confident and reproducible metabolomics studies.

Abstract Oxime reactivators are a part of the standard treatment for chemical warfare nerve agent exposure. Evaluating oxime candidates of interest in biological samples requires analytical detection methods, but oximes as a class of compounds have historically been difficult to isolate, detect, and analyze using conventional analytical techniques. Our lab previously developed novel extraction and liquid chromatography-tandem mass spectrometry (LC–MS/MS) methods to detect and quantitate 2-PAM, HI-6, HLö-7, and MMB-4 in human acetylcholinesterase knock-in, mouse carboxylesterase knock-out (KIKO) mouse plasma. These methods were validated to meet the Food and Drug Administration bioanalytical method validation requirements under Good Laboratory Practice conditions and then were utilized to analyze pharmacokinetic samples from KIKO mice. However, the future utility of these methods has been limited by the lack of reproducibly filled liquid chromatography (LC) columns. Herein we present an alternative LC column for the analysis of these oximes. Previously validated assay methods for oxime extraction from KIKO mouse plasma were evaluated for performance when analyzed by the new LC–MS/MS methodologies. Assays were evaluated for sensitivity, linearity, precision, accuracy, selectivity, and specificity. The results demonstrate that for 2-PAM, HI-6, and HLö-7, there was no change in assay performance when analyzed with the new LC–MS/MS methodologies. However, while MMB-4 was able to be retained on the new column, the sensitivity and linear range were not maintained from the original method. This study evaluates the potential of this alternative column to address inter-lot variability challenges, ensuring the robustness of oxime analytical methods.

Plant materials rich in proanthocyanidins are fractionated to determine the structure of these compounds and relate it to bioactivity. The aim of this study was to fractionate a procyanidin-rich hawthorn bark extract using low-pressure liquid chromatography and to determine the compound profile and antioxidant activity of the obtained fractions. We identified and quantified the phenolics of four fractions (I-IV) separated on a Toyopearl HW-40S column with methanol as the mobile phase, using HPLC-DAD and LC-ESI-MS techniques. The antioxidant activity was determined to comprise ABTS•+ and DPPH• scavenging activity, ferric-reducing antioxidant power (FRAP), and inhibition of β-carotene-linoleic acid emulsion oxidation. Characteristic data were subjected to principal component analysis (PCA). Fraction I contained mainly (-)-epicatechin (741.3 mg/g) and a lower amount of flavones and quercetin derivatives (100.7 mg/g). Fraction II was almost pure procyanidin B2, which accounted for 88.8% of the total phenolics. The subsequent fractions were rich in B-type procyanidin dimers, trimers, and tetramers. FRAP and antiradical activity against ABTS•+ and DPPH• of the fraction containing low-molecular weight phenolics was lower than those of the fractions with procyanidin oligomers. The antioxidant activity of fractions II-IV ranged from 8.95 to 9.28 and from 6.45 to 6.71 mmol TE/g in the ABTS and DPPH assays, respectively. Their FRAP was in the range of 17.67-21.06 mmol Fe2+/g. According to PCA, the procyanidin dimers of fractions II and III were associated with antioxidant activity in these assays. In turn, the procyanidins with the highest degree of polymerization (trimers and tetramers) present in fraction IV were related to the antioxidant activity measured in the β-carotene-linoleic acid emulsion system. Overall, the separation of purified hawthorn bark extract using low-pressure Toyopearl HW-40S column chromatography resulted in a fraction rich in procyanidin B2, as well as fractions containing procyanidins with an increasing degree of polymerization, all with high levels of antioxidant activity under various conditions and the potential for future applications in food, pharmaceuticals, and cosmetics products.

Abstract This protocol describes a trifluoroacetic acid-catalyzed regioselective 1,6-conjugate addition of mercaptopyridine and hydroxypyridine to para-quinone methides. The present reactions exhibit superior results in terms of regioselectivity, mild reaction conditions, straightforward workup (no column chromatography purification), and broad substrate scope with good functional-group compatibility. This efficient method enables the synthesis of trisubstituted methanes with pyridone and sulfide core structures.

The demand for energy-efficient light sources has witnessed a significant surge in the recent era, due to the increasing awareness towards environmental concerns and climate change. Organic Light Emitting Diodes (OLEDs) offer superior display quality, energy efficiency, and sustainability, which makes them an ideal choice for eco-conscious consumers and reduced carbon footprint-based industries. The thinner and more flexible OLEDs have revolutionized the field of smartphones, televisions, automotive displays, and wearable gadgets. Here, in this paper, an efficient OLED device is fabricated starting from the synthesis of a donor-acceptor-based emitter, followed by its purification. The organic synthesis process is explained briefly. For purification, firstly, the product mixture is purified by column chromatography, and then it is sublimed using a temperature gradient vacuum sublimation set-up to obtain a high-purity compound. The ultimate goal is to make a multi-layered OLED device that needs ultra-high-purity emitters. Making an efficient OLED needs precision and expertise; thus, showcasing a detailed step-by-step protocol is important to the science community. Here, the details about the solution-processed OLED device fabrication is unfolded in four steps (substrate cleaning, spin coating of layers, thermal evaporation of layers, and device performance measurement). First, the substrates need to be cleaned using a well-established protocol. Next, the layered architecture is obtained by spin coating and thermal deposition of desired materials to the required thickness. The thermal evaporation is done using a high vacuum thermal evaporator with an integrated glove box setup. Finally, to obtain the device's performance, measurements are to be carried out in a dark environment. This comprehensive work will provide in-depth knowledge of device fabrication and inspire more researchers towards decorating this world with OLEDs.

BackgroundDiarthron iranica (family: Thymelaeaceae), a medicinal plant native to Iran, contains a variety of beneficial phytochemicals, among which phenolic compounds with a spectrum of health-promoting activities hold a special place.ObjectivesThis study deals with the isolation and identification of the main phenolic compounds from D. iranica and investigates their inhibitory potential against α-amylase, an important enzyme in glucose metabolism, using in silico and in vitro approaches.MethodsThe purification procedure was accomplished employing chromatographic methods, including thin-layer chromatography (TLC), medium-pressure liquid chromatography (MPLC), and high-performance liquid chromatography (HPLC). The structures were determined using spectroscopic techniques: NMR (1H, 13C, DEPT), mass spectrometry (MS), and UV-Vis spectroscopy. The in vitro α-amylase inhibition was performed in triplicate across seven concentrations (0.30 - 2.80 mg/mL) using the DNS colorimetric method. Molecular docking simulations were conducted using AutoDock 4.2, with ten conformations generated per ligand.ResultsSeveral phenolic derivatives, including 5-[(β)-D-xylopyranoside-(1'''→6'')-β-D-glucopyranoside] 7-Methoxy apigenin (yuankanin, 1), 6'-Methoxy-7'-hydroxy-3'-O-7-bicoumarin (daphnoretin, 2), 4,4'-dihydroxy-3,3'-dimethoxy-7, 9’:7’, 9-diepoxylignan known as pinoresinol (3), and kusunokinin (4) with 3',4'-dimethoxy-3,4-methylenedioxydibenzyl butyrolactone structure were isolated and identified. In an α-amylase inhibition assay, compounds 1 and 3 exhibited moderate inhibitory activity with IC50 values of 1.32 mg/mL and 1.81 mg/mL, respectively, compared to the reference compound luteolin (IC50 = 0.63 mg/mL), indicating effective but relatively weaker inhibition. Compound 2 demonstrated the strongest inhibitory activity with an IC50 value of 0.71 mg/mL, surpassing compounds 1 and 3. Molecular docking studies revealed that compound 1 had a superior binding free energy of -7.13 kcal/mol, forming stable interactions through hydrogen bonding and van der Waals forces within the enzyme’s binding site. Compound 3 showed a slightly lower binding energy of -6.43 kcal/mol with fewer stabilizing interactions. However, compound 2 demonstrated poor performance in the docking assay, despite its potent inhibitory activity in the α-amylase assay.ConclusionsThe phytochemical analysis carried out on the aerial parts of D. iranica yielded the identification and characterization of four phenolic compounds, including a methoxy apigenin glycoside (1), one bicoumarin (2), and two lignans (3-4). Molecular docking studies indicated that compound 1 exhibited superior inhibitory potential compared to compound 3, with stable interactions in the enzyme’s binding site. In α-amylase inhibitory assays, these compounds displayed varying levels of activity, with compound 2 showing the highest potency (IC50 = 0.71 mg/mL), followed by compounds 1 (IC50 = 1.32 mg/mL) and 3 (IC50 = 1.81 mg/mL). However, all were less effective than the reference compound luteolin (IC50 = 0.63 mg/mL), which demonstrated superior efficacy.

An analytical method today should be both statistically reliable and enviromentally sustainable, following the principles of Green Analytical Chemistry. The official analytical method for assay and impurities in enalapril maleate tablets, as described in pharmacopoeial monographs, relies on HPLC-UV with a C8 column (250 × 4.6 mm; 5 μm) and a mobile phase consisting of acetonitrile and phosphate buffer (pH 2.2, 25 : 75 v/v). In order to reduce analysis time, cost, and environmental impact, an optimised method was developed. The chromatographic column was replaced with a C18 (100 × 4.6 mm; 3.5 μm), as the reduced particle size and shorter column length allowed for faster separations while maintaining efficiency. In addition, the mobile phase was modified to ethanol and phosphate buffer (pH 2.2, 30 : 70 v/v), since ethanol has a lower environmental impact compared with acetonitrile. Relative to the official method, the optimised procedure shortened the chromatographic run by approximately 8 minutes, reduced organic solvent consumption by 17.1 mL (equivalent to US$ 0.40) per run, and achieved better scores in the NEMI, GAPI, Eco-Scale (84 vs. 89), and AGREE (0.57 vs. 0.78) assessment tools. During validation, the linear range for enalapril was established as 140-260 μg mL-1, and the method proved capable of simultaneously quantifying enalapril, enalaprilat, and diketopiperazine with BAGI and RAPI scores of 80.0 and 67.5, respectively.