Column Chromatography Research Articles

Verbascoside and rare flavone glucosides from Citharexylum spinosum L. flowers as antihyperglycemic agents: Isolation, α-amylase inhibition, molecular docking and drug-likeness prediction

Human concern for its public health has been a cornerstone since ancient times, with humans exploring natural resources in their environment for medicinal purposes. Plants have long served as the primary source of medicinal compounds, reflecting centuries of exploration and experimentation. Today, leveraging cognitive and technological advancements, researchers continue to deepen their studies towards achieving optimal health conditions. In pursuit of antihyperglycemic phyto-alternatives, a phytochemical investigation of EtOAc and n-BuOH flower extracts of Citharexylum spinosum L. yielded four rare flavone glucosides viz. cirsilineol 4′-glucoside, jaceoside, 5,4′-dihydroxy-7,3′-dimethoxyflavone 6-glucoside and pedaliin. Additionally, the flavone aglycone apigenin, and the phenylethanoid glycoside verbascoside were isolated. The flavonoid compounds were isolated for the first time from C. spinosum tree. The extracts underwent silica gel column chromatography for simplification, followed by compound purification using preparative HPLC. Molecular structures were elucidated through NMR spectroscopic experiments, spectrometric ESI-HRMS analysis, and comparisons with existing literature data. Subsequently, extracts and isolated compounds were evaluated for their antihyperglycemic activities by assessing their ability to inhibit α-amylase enzymatic activity. EtOAc and n-BuOH extracts displayed important anti-α-amylase properties with IC50 values of 52.5 ± 1.2 and 36.3 ± 1.1 µg/mL, respectively. The results demonstrated promising inhibitory properties of the α-amylase enzyme by the polyphenols isolated in this study. IC50 values ranged from 12.8 ± 0.1 to 23.2 ± 0.1 µM for the isolated compounds, comparable to the reference standard acarbose (IC50= 12.3 ± 0.1 µM). Furthermore, a structure-activity relationship study was conducted, with all isolated molecules docked to the α-amylase enzyme (PDB: 7TAA) and their drug-likeness properties predicted.

Evaluation of hemostatic, anti-inflammatory, wound healing, skin irritation and allergy, and antimicrobial properties of active fraction from the ethanol extract of Chromolaena odorata (L.) R.M. King & H. Rob

Chromolaena odorata (L.) R.M. King & H. Rob, a perennial herb, has been traditionally utilized as a herbal remedy for treating leech bites, soft tissue wounds, burn wounds, skin infections, and dento-alveolitis in tropical and subtropical regions. The present study was to analyze the active fraction of C. odorata ethanol extract and investigate its hemostatic, anti-inflammatory, wound healing, and antimicrobial properties. Additionally, the safety of the active fraction as an external preparation was assessed through skin irritation and allergy tests. The leaves and stems of C. odorata were initially extracted with ethanol, followed by purification through AB-8 macroporous adsorption resin column chromatography to yield different fractions. These fractions were then screened for hemostatic activity in mice and rabbits to identify the active fraction. Subsequently, the hemostatic effect of the active fraction was assessed through the bleeding time of the rabbit ear artery in vivo and the coagulant time of rabbit blood in vitro. The anti-inflammatory activity of the active fraction was tested on mice ear edema induced by xylene and rat paw edema induced by carrageenin. Furthermore, the active fraction's promotion effect on wound healing was evaluated using a rat skin injury model, and skin safety tests were conducted on rabbits and guinea pigs. Lastly, antimicrobial activities against two Gram-positive bacteria (G+, Staphylococcus aureus and S. epidermidis) and three Gram-negative bacteria (G-, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa) were determined using the plate dilution method. The ethanol extract of C. odorata leaves and stems was fractionated into 30%, 60%, and 90% ethanol eluate fractions. These fractions demonstrated hemostatic activity, with the 30% ethanol eluate fraction (30% EEF) showing the strongest effect, significantly reducing bleeding time (P<0.05). A concentration of 1.0 g/mL of the 30% EEF accelerated cutaneous wound healing in rats on the 3rd, 6th, and 9th day post-operation, with the healing effect increasing over time. No irritation or allergy reactions were observed in rabbits and guinea pigs exposed to the 30% EEF. Additionally, the 30% EEF exhibited mild inhibitory effect on mice ear and rat paw edema, as well as antimicrobial activity against tested bacteria, with varying minimal inhibitory concentration (MIC) values. The 30% EEF demonstrated a clear hemostatic effect on rabbit bleeding time, a slight inhibitory effect on mice ear edema and rat paw edema, significant wound healing activity in rats, and no observed irritation or allergic reactions. Antibacterial activity was observed against certain clinically isolated bacteria, particularly the G- bacteria. This study lays the groundwork for the potential development and application of C. odorata in wound treatment.

Comparison of ELSD and RID combined with HPLC for simultaneous determination of six rare sugars in food components

Evaporative light scattering detection (ELSD) and refractive index detection (RID) are two methods commonly used in high-performance liquid chromatography (HPLC) to quantitatively analyze sugars. The performance of two detectors for the detection and quantification of six rare sugars in food samples is systematically compared in this paper. Erythritol, allulose, tagatose, mannitol, isomaltulose and trehalose were separated by HPLC on an NH2 chromatographic column using a flow gradient or isocratic composition acetonitrile–water (80:20, v/v) as mobile phase. The analytical signals for six rare sugars were obtained using ELSD and RID. The intra-day and inter-day variations were lower 3.9 % and 3.5 % for ELSD and 2.4 % and 3.6 % for RID, respectively. The limits of detection (LODs) were between 0.02 mg·mL−1 and 0.06 mg·mL−1 for ELSD in the range of 0.06–0.6 mg·mL−1. The LODs of RID were between 0.1 mg·mL−1 and 1.0 mg·mL−1 within a wider dynamic range (0.4–80 mg·mL−1). The optimized method was used to analyze seven samples. Although both methods can be used for the detection of six rare sugars, the selectivity of isomaltulose and the detection results of six rare sugars under the HPLC-RID method are not as good as the HPLC-ELSD method. Therefore, the HPLC-ELSD method is more suitable for the detection of six rare sugars in food.

Polymeric monolithic columns based on natural wood for rapid purification of targeted protein

With multiscale hierarchical structure, wood is suitable for a range of high-value applications, especially as a chromatographic matrix. Here, we have aimed to provide a weak anion-exchange polymeric monolithic column based on natural wood with high permeability and stability for effectively separating the targeted protein. The wood-polymeric monolithic column was synthesized by in situ polymerization of glycidyl methacrylate and ethylene glycol dimethacrylate in wood, and coupled with diethylaminoethyl hydrochloride. The wood-polymeric monolithic column can be integrated with fast-protein liquid chromatography for large-scale protein purification. According to the results, the wood-polymeric monolithic column showed high hydrophilicity, permeability and stability. Separation experiments verified that the wood-polymeric monolithic column could purify the targeted protein (spike protein of SARS-COV-2 and ovalbumin) from the mixed proteins by ion exchange, and the static adsorption capacity was 33.04 mg mL−1 and the dynamic adsorption capacity was 24.51 mg mL−1. In addition, the wood-polymerized monolithic column had good stability, and a negligible decrease in the dynamic adsorption capacity after 20 cycles. This wood-polymerized monolithic column can provide a novel, efficient, and green matrix for monolithic chromatographic columns.

Preparation and Characterization of Stationary Phase Gradients on C8 Liquid Chromatography Columns

Continuous C8 stationary phase gradients are created on commercial Waters Symmetry Shield RP8 columns by strategically cleaving the C8 moieties in a time-dependent fashion. The method relies on the controlled infusion of a trifluoracetic acid/water/acetonitrile solution through the column to cleave the organic functionality (e.g., C8) from the siloxane framework. The bond cleavage solution is reactive enough to cleave the functional groups, even with polar groups embedded within the stationary phase to protect the silica. Both the longitudinal and radial heterogeneity were evaluated by extruding the silica powder into polyethylene tubing and evaluating the percent carbon content in the different sections using thermogravimetric analysis (TGA). TGA analysis shows the presence of a stationary phase gradient in the longitudinal direction but not in the radial direction. Two different gradient profiles were formed with good reproducibility by modifying the infusion method: one exhibited an ‘S’-shaped gradient while the other exhibited a steep exponential-like gradient. The gradients were characterized chromatographically using test mixtures, and the results showed varied retention characteristics and an enhanced ability to resolve nicotine analytes.

Antidiabetic and antihyperlipidemic effects of crude fractions from Chlorophytum borivilianum root methanolic extract on streptozotocin induced diabetic rats and phytochemical investigation by LCMS analysis.

To evaluate the in vitro and in vivo pharmacological efficacy of the plant Chlorophytum borivilianum in diabetes and hyperlipidemia and to confine and describe the synthetic constituents from the roots that are in charge of the action. The present study was carried out to investigate the ethno-medical use of Chlorophytum borivilianum root methanolic extract as a potential anti-diabetic and antihyperlipidemic agent in STZ-induced diabetic rats. Extract was tested for in vitro and in vivo biological activities. Soxhlet extraction was carried out using methanol as a solvent, and TLC and column chromatography were used for fractionation. Liquid Chromatography and Mass Spectroscopic study confirmed the structures of isolated compounds. Chlorophytum borivilianum root methanolic extract showed the presence of phytoconstituents as Dihydrocapsaicin, Reserpine, Deserpidine, Biliverdin-IX-α, and Cassiamin C having a therapeutic effect. Dihydrocapsaicin was identified at RT 7.572 and the Chlorophytum borivilianum root chloroform methanolic extract fraction noticeably depleted increased blood glucose levels and had positive effects on altered lipid profile after administering a dose of 150 mg/kg orally compared with oral hypoglycemic drug metformin. All the results are dose-dependent. Active chloroform-methanol fraction from methanol extract showed the presence of anti-diabetic compound, Dihydrocapsaicin. The chloroform-methanol fraction from the methanolic extract of Chlorophytum borivilianum root can inhibit the parameters linked to diabetes and hyperlipidemia.

Comparison of reversed-phase, hydrophilic interaction, and porous graphitic carbon chromatography columns for an untargeted toxicometabolomics study in pooled human liver microsomes, rat urine, and rat plasma

IntroductionUntargeted metabolomics studies are expected to cover a wide range of compound classes with high chemical diversity and complexity. Thus, optimizing (pre-)analytical parameters such as the analytical liquid chromatography (LC) column is crucial and the selection of the column depends primarily on the study purpose.ObjectivesThe current investigation aimed to compare six different analytical columns. First, by comparing the chromatographic resolution of selected compounds. Second, on the outcome of an untargeted toxicometabolomics study using pooled human liver microsomes (pHLM), rat plasma, and rat urine as matrices.MethodsSeparation and analysis were performed using three different reversed-phase (Phenyl-Hexyl, BEH C18, and Gold C18), two hydrophilic interaction chromatography (HILIC) (ammonium-sulfonic acid and sulfobetaine), and one porous graphitic carbon (PGC) columns coupled to high-resolution mass spectrometry (HRMS). Their impact was evaluated based on the column performance and the size of feature count, amongst others.ResultsAll three reversed-phase columns showed a similar performance, whereas the PGC column was superior to both HILIC columns at least for polar compounds. Comparing the size of feature count across all datasets, most features were detected using the Phenyl-Hexyl or sulfobetaine column. Considering the matrices, most significant features were detected in urine and pHLM after using the sulfobetaine and in plasma after using the ammonium-sulfonic acid column.ConclusionThe results underline that the outcome of this untargeted toxicometabolomic study LC-HRMS metabolomic study was highly influenced by the analytical column, with the Phenyl-Hexyl or sulfobetaine column being the most suitable. However, column selection may also depend on the investigated compounds as well as on the investigated matrix.Graphical abstract

ISOLATION OF SQUALENE FROM DICHLOROMssETHANE EXTRACT OF CROTON NIGRITANUS LEAF

A precursor of steroids with a variety of biological activities is squalene. The purpose of the study was to identify and describe the industrial/medical chemical found in Croton nigritanus. Using the solvents such as; dichloromethane, ethyl acetate, and n-hexane, the procedure entails a liquid-liquid extraction stage using the partition method and a solid-liquid extraction step using the maceration method. For separation and purification, gravity column chromatography (GCC) and thin-layer chromatography were employed. The isolated chemical's identity was determined using FTIR, 1H NMR and GC-MS spectroscopy. The inquiry successfully extracted and analyzed the major chemical compound which was identified as Squalene.

Online Two-Dimensional Liquid Chromatography for Determination of Selected Flavonoids in Wine Samples

The development of two-dimensional chromatography systems is crucial in many scientific fields as it provides improved separation efficiency and selectivity of complex mixtures. By separating compounds based on two independent properties, these systems can offer a more detailed understanding of sample composition. This increased level of detail is particularly useful in industries such as pharmaceuticals, environmental analysis, and proteomics, where complex mixtures with similar properties need to be differentiated. A new chromatography system with two dimensions was developed to separate and quantify specific flavonoids in German wine samples. The first dimension used a homemade hydrophilic interaction liquid chromatographic column, while the second dimension used a reversed-phase liquid chromatographic column. This 2D ZIC-HILIC x RP technique was able to address the problem of peak overlapping that commonly occurs in one-dimensional liquid chromatography, especially for natural products. The study presents analytical and statistical data for both techniques, and suggests that the use of two-dimensional chromatography can improve the resolution of conventional one-dimensional liquid chromatography. In a conventional ZIC-HILIC-HPLC system, apigenin and naringenin were detected together but were successfully separated and measured using a 2DLC system. The limits of detection (LOD) for apigenin and naringenin were found to be 0.0705 µg/mL and 0.1300 µg/mL, respectively, while the limits of quantification (LOQ) were 0.2136 µg/mL and 0.3939 µg/mL, respectively. The recovery rate for both compounds was over 99%, with a relative standard deviation (RSD) of less than 1.5%. The linearity range for both compounds was between 0.005 µg/mL to 10 µg/mL, and the correlation coefficient (r2) was determined to be 0.9998. For hesperidin and rutin, the LOD and LOQ were found to be 0.0336 µg/mL and 0.1018 µg/mL, and 0.0120 µg/mL and 0.0363 µg/mL, respectively. The linearity range for both compounds was between 0.005 µg/mL to 12 µg/mL, and the correlation coefficient (r2) was determined to be 0.9999.

Structural characterization and improves cognitive disorder in ageing mice of a glucomannan from Dendrobium huoshanense

In the present work, a neutral polysaccharide (DHP-2W) with attenuating cognitive disorder was identified from Dendrobium huoshanense and its structure was clarified. The polysaccharide was successfully purified from D. huoshanense by column chromatography and its activity was evaluated. With a molecular weight of 508.934kDa, this polysaccharide is composed of mannose and glucose at a molar ratio of 75.81: 24.19. Structural characterization revealed that DHP-2W has a backbone consisting of 4)-β-D-Manp-(1 and 4)-β-D-Glcp-(1. In vivo experiments revealed that DHP-2W improved cognitive disorder in D-galactose treated mice and relieved oxidative stress and inflammation. DHP-2W attenuates D-galactose-induced cognitive disorder by inhibiting the BCL2/BAX/CASP3 pathway and activating the AMPK/SIRT pathway, thereby inhibiting apoptosis. Furthermore, DHP-2W had a significant effect on regulating the serum levels of Flavin adenine dinucleotide, Shikimic acid, and Kynurenic acid in aged mice. These, in turn, had a positive impact on AMPK/SIRT1 and BCL2/BAX/CASP3, resulting in protective effects against cognitive disorder.

A comparative study on the synergistic activities of fractions and crude essential oil of Syzygium aromaticum

BackgroundSyzygium aromaticum has been used for over a century because it has biological qualities for several health-related problems, and the crude essential oils are known to contain several components that could be responsible for synergistic or antagonistic potentials in relation to their biological uses. The study was designed to compare the biological activities of the crude essential oil from the bud of Syzygium aromaticum and fractions obtained from column chromatography.Materials and methodsThe essential oil was extracted by hydrodistillation, and two prominent fractions, C1 and C2, were obtained from the column chromatography separation of the crude essential oil. The quantitative phytochemical analysis was done by standard methods, the antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging assays. The antifungal activities were assessed by the agar diffusion method, while the anti-inflammatory activity was done using a lipoxygenase inhibitory assay. The major component observed in the crude and fractions was determined by gas chromatography–mass spectrometry (GC–MS) analysis.ResultsThe phytochemical analysis showed that phenolic acid, tannin, saponin, flavonoids, alkaloids, cardiac glycosides, and terpenoids were present in variable quantities in all of the fractions and crude essential oil. The half-maximal inhibitory concentration (IC50) value for DPPH scavenging activity ranged from 226.1 to 301.2 µg/mL while that of ABTS scavenging activity ranged from 211.0 to 316.0 µg/mL. The lowest value of IC50 for both DPPH and ABTS was observed in fraction C1. The crude essential oil with a minimum inhibitory concentrations (MIC) value of 8.00 mg/mL was the most efficient against Candida albicans, Aspergillus penicillum, and Aspergillus niger. The anti-inflammatory assay showed C1 to have the lowest value of IC50 for the lipoxygenase assay. The major component observed in the crude essential oil as well as fraction C1 was eugenol, while fraction C2 had β-caryophyllene as the major component.ConclusionsThe result showed that all fractions as well as the crude essential oil exhibited antioxidant, antifungal, and anti-inflammatory activities, and the crude essential oil was synergic only with respect to its antifungal activity, while fraction C1 was more efficient as an anti-inflammatory and antioxidant agent.

Новий метаболіт Myrothecium cinctum 910 трихотеценової природи та його біологічні властивості

Fungi of the Myrothecium genus are well-known producers of macrocyclic trichothecenes, characterized by the acute toxic effect on warm-blooded organisms and fungicidal action against a wide range of fungi. At the same time, as established by us earlier, M. cinctum (current name Striaticonidium cinctum) 910 shows a wide spectrum of antibacterial activity: along with antifungal activity against mycelial, yeast-like and phytopathogenic test-cultures it strongly inhibited the growth of gram-positive test-strains and to lesser extend – the growth of gram-negative and phytopathogenic bacteria. This strain also revealed significant phytotoxic potential suppressing the growth of green algae Chlorella strains. The aim of the work was to isolate, to purify and to obtain in crystalline form the biological active metabolites of M. cinctum 910 and to study their physicochemical characteristics and biological activity. Methods. To obtain biologically active metabolites in the purified crystalline form, the extraction followed by separation by column chromatography and recrystallization was used; physicochemical methods and microbiological tests were used for characterization of their properties. Results. Three substances which showed the wide spectrum of biological activity to indicator test-cultures were obtained in crystalline form: preparation МС910 with high antibiotic activity and preparations from fractions 8/2 and 9 with high antifungal and phytotoxic activities. Taking into account the data of spectroscopy in the UV-range of spectrum, of spectrums of antibiotic activity and of qualitative reaction with 4-(4-nitrobenzyl)pyridine, it can be concluded that active fractions 8/2 and 9 contain macrocyclic trichothecenes whereas the metabolite МС910 (which is proved by data of IR-spectroscopy) belongs to simple trichothecenes – substances like to macrocyclic trichothecenes but without macrocyclic fragment in their structure. Conclusions. Antibiotic properties of M. cinctum 910 are due to the complex of biological active metabolites with different biological and physicochemical properties. In view on the biological activity of MC910, it can be argued that it has atypical for trichothecenes, including macrocyclc trichothecenes, antibacterial properties as well as it is non-toxic and does not reveal dermatocidal activity in studied concentrations. Therefore, we assume that MC910 is new, not described earlier metabolite of trichothecene nature, which needs further studies. At the same time, active metabolites from fractions 8/2 and 9 are represented by macrocyclic trichothecenes, which is also indicated by the spectra of their biological activity.

In vitro cytotoxic activity on KATO-III cancer cell lines of mangiferolic acid purified from Thai Tetragonula laeviceps propolis

Gastric cancer is a global health concern, but current treatment with chemotherapy and surgery is often inadequate, prompting the exploration of alternative treatments. Propolis is a natural substance collected by bees known for its diverse properties linked to floral sources. The Dichloromethane Partitioned Extract (DPE) from Tetragonula laeviceps propolis, in Bankha district, Thailand was previously shown to possess significant cytotoxicity against KATO-III gastric cancer cells, while showing lower cytotoxicity toward WI-38 normal fibroblast cells. Here, the DPE was further fractionated by column chromatography, identified active fractions, and subjected to structural analysis using nuclear magnetic resonance spectroscopy. Cytotoxicity against KATO-III cells was reevaluated, and programmed cell death was analyzed using flow cytometry. Expression levels of cancer-related genes were measured using quantitative real-time reverse transcriptase PCR. Cardol C15:2 (compound 1) and mangiferolic acid (MF; compound 2) were discovered in the most active fractions following structural analysis. MF exhibited strong cytotoxicity against KATO-III cells (IC50 of 4.78–16.02 μg/mL), although this was less effective than doxorubicin (IC50 of 0.56–1.55 μg/mL). Morphological changes, including decreased cell density and increased debris, were observed in KATO-III cells treated with 30 μg/mL of MF. Significant induction of late-stage apoptosis and necrosis, particularly at 48 and 72 h, suggested potential DNA damage and cell cycle arrest, evidenced by an increased proportion of sub-G1 and S-phase cells. Doxorubicin, the positive control, triggered late apoptosis but caused more necrosis after 72 h. Furthermore, MF at 30 μg/mL significantly increased the expression level of COX2 and NFκB genes linked to inflammation and cell death pathways. This upregulation was consistent at later time points (48 and 72 h) and was accompanied by increased expression of CASP3 and CASP7 genes. These findings suggest MF effectively induces cell death in KATO-III cells through late apoptosis and necrosis, potentially mediated by upregulated inflammation-related genes.

Computational investigation of 2, 4-Di Tert Butyl Phenol as alpha amylase inhibitor isolated from Coccinia grandis (L.) Voigt using molecular docking, and ADMET parameters

IntroductionDiabetes Mellitus is the metabolic disorder most prevalent globally, accounting for a substantial morbidity rate. The conventional drugs available for the management of diabetes are either expensive or lack the required efficacy. The purpose of this research is to isolate and characterize an active phytoconstituent from Coccinia grandis and assess its anti-diabetic properties. Methods and materialsStems of Coccinia grandis are subjected to successive extraction and isolation. The isolated compound by column chromatography was characterized by FTIR (fourier-transform infrared), 1 H NMR (proton nuclear magnetic resonance), and Mass spectroscopy. The antidiabetic potential of the isolated compound was evaluated by in-vitro alpha-amylase inhibitory activity. Further, the compound was subjected to molecular docking studies to study its interaction with the human pancreatic alpha-amylase (Molegro Virtual Docker) as well to determine the pharmacokinetic and toxicity profile using computational techniques (OSIRIS property explorer, Swiss ADME, pkCSM, and PreADMET). ResultsThe characterization of the compound suggests the structure to be 2,4-ditertiary butyl phenol. The in-vitro alpha-amylase inhibitory study indicated a concentration-dependent inhibition and the IC50 (median lethal dose) value of the isolated compound was found to be 64.36 μg/ml. The docking study with the A chain of receptor 5EMY yielded a favorable docking score of −81.48 Kcal mol−1, suggesting that the compound binds to the receptor with high affinity through electrostatic, hydrophobic, and hydrogen bonds. Furthermore, the silico ADME analysis of the compound revealed improved metabolism, a skin permeability of −3.87 cm/s, gastrointestinal absorption of 95.48 %, and a total clearance of 0.984 log ml min−1 kg−1. In silico toxicity analysis also predicted cutaneous irritations but no carcinogenicity, mutagenicity, or hepatotoxicity. ConclusionThe data suggested that the isolated compound (2, 4-tertiary butyl phenol) has the potential to inhibit the alpha-amylase activity and possess optimal ADME properties as well as tolerable side effects.

Construction of Hierarchical Porous UiO-66-Br2@PS/DVB-Packed Columns by High Internal Phase Emulsion Strategy for Enhanced Separation of CF4/N2 and SF6/N2.

Recovery and separation of anthropogenic emissions of electronic specialty gases (F-gases, such as CF4 and SF6) from the semiconductor sector are of critical importance. In this work, the hierarchical porous UiO-66-Br2@PS/DVB-packed column was constructed by a high internal phase emulsions strategy. UiO-66-Br2@PS/DVB exhibits a superior selectivity of CF4/N2 (2.67) and SF6/N2 (3.34) predicted by the IAST due to the diffusion limitation in the micropore and the gas-framework affinity. Especially, UiO-66-Br2@PS/DVB showed significant CF4 and SF6 retention and enabled the successful separation of CF4/N2 and SF6/N2 with a resolution of 2.37 and 8.89, respectively, when used as a packed column in gas chromatography. Compared with the Porapak Q column, the HETP of the UiO-66-Br2@PS/DVB-packed column decreased and showed good reproducibility. This research not only offers a convenient method for fabricating a hierarchical porous MOF-packed column but also showcases the prospective utilization of MOFs for the separation of the F-gas/N2 mixture.

Unveiling the Untapped Potential of Bertagnini's Salts in Microwave-Assisted Synthesis of Quinazolinones.

Microwave-assisted organic synthesis (MAOS) has emerged as a transformative technique in organic chemistry, significantly enhancing the speed, efficiency, and selectivity of chemical reactions. In our research, we have employed microwave irradiation to expedite the synthesis of quinazolinones, using water as an eco-friendly solvent and thereby adhering to the principles of green chemistry. Notably, the purification of the product was achieved without the need for column chromatography, thus streamlining the process. A key innovation in our approach is using aldehyde bisulfite adducts (Bertagnini's salts) as solid surrogates of aldehydes. Bertagnini's salts offer several advantages over free aldehydes, including enhanced stability, easier purification, and improved reactivity. Green metrics and Eco-Scale score calculations confirmed the sustainability of this approach, indicating a reduction in waste generation and enhanced sustainability outcomes. This methodology facilitates the synthesis of a diverse array of compounds, offering substantial contributions to the field, with potential for widespread applications in pharmaceutical research and beyond.

Forced Degradation Products of Olmesartan Medoxomil and their In Vitro Genotoxicity Assessment by Comet Assay using HEK Cells

AbstractThis study designed to characterize major forced degradation products of Olmesartan Medoxomil (OM) and to perform genotoxicity profiling using acid, base, and hydrogen peroxide‐induced stress degradation studies. The isolation of degradation products was conducted on flash chromatography using FlashPure ECOFLEX C18 4 g column with gradient elution and diode array detection‐based fraction collection, and characterized by mass spectrometry. Three impurities (impurity‐1, 2, and 3) were identified with respective m/z values of 447.213, 461.229, and 463.208. The genotoxic potential of impurities was assessed by in silico prediction, and in vitro comet assay using HEK cells. The extent of DNA damage was analyzed using Comet Assay IV imaging software. For impurity‐1, head and tail lengths and their respective total intensities were significantly (p&lt;0.001) larger than those of dimethyl sulfoxide (solvent) control and comparable to positive control. Other parameters such as mean grey level and total area of comet were statistically significant as compared with the solvent and positive controls. Overall, impurity‐1 was a possible genotoxic impurity and a known major degradation product in OM tablet dosage form. In silico prediction also revealed impurity‐1 as toxicity class 3, whilst impurities 2 and 3 were of toxicity class 4.

Anti-Photodamage Effect of Agaricus blazei Murill Polysaccharide on UVB-Damaged HaCaT Cells.

UVB radiation is known to induce photodamage to the skin, disrupt the skin barrier, elicit cutaneous inflammation, and accelerate the aging process. Agaricus blazei Murill (ABM) is an edible medicinal and nutritional fungus. One of its constituents, Agaricus blazei Murill polysaccharide (ABP), has been reported to exhibit antioxidant, anti-inflammatory, anti-tumor, and immunomodulatory effects, which suggests potential effects that protect against photodamage. In this study, a UVB-induced photodamage HaCaT model was established to investigate the potential reparative effects of ABP and its two constituents (A1 and A2). Firstly, two purified polysaccharides, A1 and A2, were obtained by DEAE-52 cellulose column chromatography, and their physical properties and chemical structures were studied. A1 and A2 exhibited a network-like microstructure, with molecular weights of 1.5 × 104 Da and 6.5 × 104 Da, respectively. The effects of A1 and A2 on cell proliferation, the mitochondrial membrane potential, and inflammatory factors were also explored. The results show that A1 and A2 significantly promoted cell proliferation, enhanced the mitochondrial membrane potential, suppressed the expression of inflammatory factors interleukin-1β (IL-1β), interleukin-8 (IL-8), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α), and increased the relative content of filaggrin (FLG) and aquaporin-3 (AQP3). The down-regulated JAK-STAT signaling pathway was found to play a role in the response to photodamage. These findings underscore the potential of ABP to ameliorate UVB-induced skin damage.

Structural characterization of the polysaccharide from the black crystal region of Inonotus obliquus and its effect on AsPC-1 and SW1990 pancreatic cancer cell apoptosis

In this study, one water soluble polysaccharide (IOP1–1) with a weight average molecular weight of 6886 Da was obtained from the black crystal region of Inonotus obliquus by hot water extraction, DEAE-52 cellulose extraction and Sephadex-100 column chromatography purification. Structural analysis indicated that IOP1–1 was a glucan with a main chain composed of α-Glcp-(1 → 4)-α-Glcp-(1 → 4)-β-Glcp-(1 → 4)-β-Glcp-(1 → 4)-α-Glcp-(1 → 6)-β-Glcp-(1 → 4)-α-Glcp-(1 → 3)-β-Glcp-(1→. The CCK-8 assay results showed that IOP1–1 inhibited AsPC-1 and SW1990 pancreatic cancer cell proliferation in a concentration-dependent manner. Flow cytometric analysis revealed that IOP1–1 induced cell cycle arrest in AsPC-1 and SW1990 cells. Hoechst 33342 staining and Annexin V-FITC/PI double staining analysis showed that IOP1–1 could induce apoptosis in AsPC-1 and SW1990 cells. Furthermore, western blot analysis confirmed that IOP1–1 could induce apoptosis in AsPC-1 and SW1990 pancreatic cancer cells through three pathways: the mitochondrial pathway, the death receptor pathway, and endoplasmic reticulum stress. According to these research data, IOP1–1 may be utilized as an adjuvant treatment to anticancer medications, opening up new application prospects and opportunities.

Numerical Approximation of a Nonequilibrium Model of Gradient Elution Chromatography Considering Different Functional Relationships between Model Parameters and Solvent Composition.

In this paper, a rigorous theoretical study is conducted to analyze the influence of varying solvent compositions on the retention characteristics of elution profiles within a fixed-bed liquid chromatographic column. In gradient chromatography, the propagation speed of elution profiles is manipulated through a progressive variation in the mobile-phase composition. Consequently, enhanced separation of the mixture components can be achieved together with a reduction in the requisite recycling times for subsequent injections. In other words, both the efficiency and the selectivity of the column can be enhanced. The lumped kinetic model coupled with the convection-diffusion equation for the volume fraction of the solvent is applied to simulate the process. The resulting nonlinear model equations are numerically solved by applying a semidiscrete second-order finite-volume method. The numerical solutions are utilized to quantify the effects of gradient starting and ending times, solvent composition, solvent strength parameters, and gradient slope on the concentration profiles. Additionally, temporal numerical moments are plotted versus the starting and ending times of the gradient, and standard performance criteria are presented for evaluating the process performance. The outcomes of this investigation will contribute to further enhancements in gradient elution chromatography.