Column Chromatography Research Articles

Isolation, characterization, and analysis of pure compounds from Leea macrophylla leaf extract for antibacterial, antidiabetic, cytotoxicity and phytotoxicity

This research aims to isolate, characterize, and analyze pure compounds from Leea macrophylla leaf extract to investigate its antibacterial, antidiabetic, cytotoxic, and phytotoxic effects. Fresh leaves were collected, dried, and subjected to methanol extraction to obtain a crude extract. From the petroleum ether fraction (PEF) of this extract, three fractions—designated LM1, LM2, and LM3—were prepared using column chromatography. The fractions were tried to be characterized in search for single compound by instrumental technique like ATR-FTIR, 1H NMR and 13C NMR but the 1H NMR and 13C NMR spectra were found complex which were difficult to interpret. To dispel the doubt and get clear idea about the structure, GC-MS analysis of the compounds was carried out whose result showed that all the three extracts were decomposed to several small organic compounds that made the structure elucidation difficult. For this complication, the characterization of the extracts was not possible. Numerous compounds were identified in the methanol extract of L.macrophylla through GC-MS analysis. Among these compounds, Benzene, 1,2,3-trimethyl- and Undecane were found in higher percentages in LM1. LM2 contained Azulene and Bicyclo [4.4.1]undeca-1,3,5,7,9-pentaene, while LM3 was characterized by the presence of 9,9-Dimethoxybicyclo [3.3.1]nona-2,4-dione and 11-(2-Cyclopenten-1-yl)undecanoic acid, among others. The antibacterial activity of these fractions was evaluated against various bacterial strains, demonstrating broad-spectrum effectiveness. LM1 fraction showed the highest antibacterial activity against Proteus sp. With zone of inhibition 25 mm and weak activity against S. sonnei with zone of inhibition 5 mm. LM2 showed the highest activity to both E. cocci and P. aeruginosa with the zone of inhibition of 18 mm and comparatively lower but significant against Proteus sp. LM3 was highly active to S. sonnei with zone of inhibition 20 mm and lower but quite significant against Proteus sp. Moreover, the anti-diabetic potential was assessed, with LM1 showing the strongest α-amylase inhibitory activity, outperforming quercetin (standard). The IC50 values of LM1, LM2, LM3, and quercetin were 57.36 μg/mL, 100.66 μg/mL, 164.92 μg/mL, and 97.45 μg/mL, respectively. In addition, cytotoxicity was assessed using a brine shrimp lethality bioassay, and phytotoxicity was evaluated through seed germination and growth assays. The results suggest that L.macrophylla leaf extracts have potential applications in antimicrobial, antidiabetic, and anti-cancer contexts. This comprehensive study bridges gaps in knowledge surrounding L.macrophylla's multifaceted properties, offering insights into its therapeutic and ecological potential for healthcare and environmental management.

Multicomponent Synthesis of Structurally Diverse Spiroheterocycles using Bio-organic Catalyst in Aqueous Medium

Background: MCRs are one of the most significant tools in the synthesis of organic compounds. MCR is a rapid chemical technique that uses three or more reactants to produce products that sustain all structural and substructural properties of the initial components. MCRs are useful in all fields of synthetic chemistry because of their rapid rate of reaction, simple procedure and excellent yields. We reported an efficient and environmentally friendly domino approach for the synthesis of spiroheterocycles spiro annulated with indeno[1,2-b]quinoline. Method: The spiroheterocycles with privileged heterocyclic substructures have been synthesized using taurine (2-aminoethanesulfonic acid) as a green, sustainable, bio-organic and recyclable catalyst in a three-component reaction of isatins, 1,3-diketones, and 1-napthylamine in aqueous media. The present synthetic method is probably the first report to synthesize spiroheterocycles, spiroannulated with indeno[1,2-b]quinoline. Furthermore, the approach is valuable because of the excellent yield that results from the reaction in 15-20 min. Result: The optimization of reaction conditions is an important case of efficient synthesis. The solvent, temperature, time and catalyst loading were all examined. The reusability of the catalyst was also investigated experimentally. The used catalyst taurine has a high activity as well as good reusability. The present synthetic protocol will be extended to synthesise a library of hybrid compounds. The present synthetic approach is cost-effective, and time-efficient with an easy-workup methodology that gives outstanding yields (80–95%) in 15–20 min. Conclusion: Taurine-catalyzed multicomponent reaction is a novel and efficient method for the synthesis of spiroannulated indeno[1,2-b]quinolines. The high catalytic activity of taurine as a catalyst with water as a green solvent makes the process environmentally friendly. The special features of the synthetic protocol include synthetic efficiency, operational simplicity, and reusability of the catalyst and it is expected to make significant contributions not only to drug discovery studies but also to pharmaceutical and therapeutic chemistry in view of introducing molecular diversity in the synthesized molecules.

Ultrasound-assisted extraction of triterpenoids from Chaenomeles speciosa leaves: Process optimization, adsorptive enrichment, chemical profiling, and protection against ulcerative colitis.

For the valorization of Chaenomeles speciosa leaves, this study focused on extraction, enrichment, chemical profiling, and investigation of the biological activity of its abundant triterpenoid components. Initially, the total triterpenoids in C. speciosa leaves were extracted by ultrasonic-assisted extraction (UAE) method, with the extraction process optimized through response surface methodology (RSM). Under the optimal conditions of extraction solvent 93% EtOH, ultrasound power 390W, extraction time 30min, extraction temperature 70°C, liquid-to-solid ratio 25mL/g, and 2 extraction cycles, the maximum total triterpenoids yield (TTY) reached 36.77±0.40mg/g. The total triterpenoids in the crude extract were subsequently enriched by X-5 resin column chromatography, resulting in a fourfold increase in purity, reaching 73.27±0.84%. Thirteen compounds in the triterpenoid-rich fraction (TRF) were identified through UPLC-QTOF-MS/MS, and five major triterpenoids (oleanolic acid, ursolic acid, betulinic acid, maslinic acid, and pomolic acid) were simultaneously quantified by HPLC-QQQ-MS. Furthermore, TRF demonstrated a notable amelioration against dextran sodium sulfate (DSS)-induced ulcerative colitis in mice, indicating its promise as a potent intervention for this condition. In summary, this study will contribute to enhancing the utilization efficiency of Chaenomeles speciosa leaves.

Antitrypanosomal activity of Crossopteryx febrifuga and phytochemical profiling using LC-MS/MS analysis coupled to molecular network and SIRIUS

Crossopteryx febrifuga (Rubiaceae) is a plant widely used in traditional African medicine to treat tryapnosomiasis. The aim of our study was to evaluate the antitrypanosomal activity of C. febrifuga extracts and to identify the compounds responsible for this activity. We prepared 4 extracts by successive maceration of plant leaf powder in n-hexane, dichloromethane, methanol and water. The antitrypanosomal activity of the extracts were assessed on Trypanosoma brucei brucei and their selectivity on Leishmania mexicana mexicana and human non cancer WI38 fibroblast cells. The dichloromethane extract, the most antitrypanosomal (IC50 of 9.3 ± 0.8 μg/mL) was fractionated on an Open Column Chromatography to give 14 fractions. Fractions 6–9 were the most active with an IC50 ranging from 1.3 to 2.1 μg/mL. All fractions were analyzed by UPLC-ZenoTOFHRMS, followed by manual dereplication of metabolites detected in the most active fractions. Manual dereplication was aided by the Molecular Network (MN) and SIRIUS. Metabolic profiling of fractions 6–9 has enabled us to identify 33 compounds, most of which were reported for the first time in C. febrifuga. These include buddlenol C (6), naringenin (7), maslinic acid (22), corosolic acid or isomer (24), asperphenamate (25), hydroxyoctadecenoic acid (29), sumaresinolic acid or isomer I and II (30 et 31), glycyrrhetinic acid (32) and oleanolic acid (36). The bioactivity-based approach identified maslinic (22), corosolic (24), and oleanolic (36) acids as linked to the antitrypanosomal activity. The data obtained support the traditional use of C. febrifuga in the traditional treatment of trypanosomiasis. Further studies are required to verify the activity observed in vivo.

The Inhibitory Effects of the Herbals Secondary Metabolites (7α-acetoxyroyleanone, Curzerene, Incensole, Harmaline, and Cannabidiol) on COVID-19: A Molecular Docking Study.

Since the COVID-19 outbreak in early 2020, researchers and studies are continuing to find drugs and/or vaccines against the disease. As shown before, medicinal plants can be very good sources against viruses because of their secondary compounds which may cure diseases and help in survival of patients. There is a growing trend in the filed patents in this field. In the present study, we test and suggest the inhibitory potential of five herbal based extracts including 7α-acetoxyroyleanone, Curzerene, Incensole, Harmaline, and Cannabidiol with antivirus activity on the models of the significant antiviral targets for COVID-19 like spike glycoprotein, Papain-like protease (PLpro), non-structural protein 15 (NSP15), RNA-dependent RNA polymerase and core protease by molecular docking study. The Salvia rythida root was extracted, dried, and pulverized by a milling machine. The aqueous phase and the dichloromethane phase of the root extractive were separated by two-phase extraction using a separatory funnel. The separation was performed using the column chromatography method. The model of the important antivirus drug target of COVID-19 was obtained from the Protein Data Bank (PDB) and modified. TO study the binding difference between the studied molecules, the docking study was performed. These herbal compounds are extracted from Salvia rhytidea, Curcuma zeodaria, Frankincense, Peganum harmala, and Cannabis herbs, respectively. The binding energies of all compounds on COVID-19 main targets are located in the limited area of 2.22-5.30 kcal/mol. This range of binding energies can support our hypothesis for the presence of the inhibitory effects of the secondary metabolites of mentioned structures on COVID-19. Generally, among the investigated herbal structures, Cannabidiol and 7α- acetoxyroyleanone compounds with the highest binding energy have the most inhibitory potential. The least inhibitory effects are related to the Curzerene and Incensole structures by the lowest binding affinity. The general arrangement of the basis of the potential barrier of binding energies is in the order below: Cannabidiol > 7α-acetoxyroyleanone > Harmaline> Incensole > Curzerene. Finally, the range of docking scores for investigated herbal compounds on the mentioned targets indicates that the probably inhibitory effects on these targets obey the following order: main protease> RNA-dependent RNA polymerase> PLpro> NSP15> spike glycoprotein.

Extraction of Fucoxanthin from Marine Organisms and Its Therapeutic Effect on Cancer Cells

As China’s economy develops and its population grows, resources are becoming increasingly scarce. As a result, attention is shifting towards utilising marine resources, and the ability to process them is an increasingly important area of development.Within the traditional marine industry, kelp is primarily used for fucoidan extraction, with other substances such as kelp pigments often cast off as waste. Fucoxanthin constitutes the major carotenoid component found within kelp pigments. These pigments are believed to possess immunomodulatory, anti-inflammatory, and anti-tumour properties, and are of multifaceted benefit to human health by counteracting cellular oxidative damage induced by free radicals. Kelp served as the raw material in this study. Macroporous resin was utilized to extract Fucoxanthin from the kelp extract. Only an aqueous ethanol solution was employed for the extraction process. Subsequently, reduced-pressure column chromatography was conducted to quickly enrich the Fucoxanthin. Finally, the anti-tumour activity and free radical scavenging ability of the Fucoxanthin were analyzed. The results of the experiment revealed that Fucoxanthin can be effectively adsorbed by X-5 resin and its adsorption capability is robust. Additionally, using a 60% concentration of ethanol solution can promptly and effectively elute the absorbed Fucoxanthin from the X-5 resin. Moreover, after incubation for 48 hours at a concentration of 80 μg/mL of Fucoxanthin, the expression levels of p65, p52, and p50 genes significantly decreased in cancer cells. Fucoxanthin, a pigment found in kelp, exhibits potent antioxidant activity. The findings suggest that Fucoxanthin possesses the ability to effectively suppress the expression of cancer cells and scavenge free radicals.

Physicochemical properties, antioxidant and anti-inflammatory activities of polysaccharides from lotus leaves by pectinase-assisted extraction

In order to improve the extraction yield of lotus leaf polysaccharide (LLP) and offer scientific foundation for utilization of LLPs in the food and pharmaceutical industries, pectinase-assisted extraction method was employed for the preparation of LLP. The chemical composition, physicochemical properties (e.g. rheological and thermal properties etc.), antioxidant and anti-inflammatory effects of LLP and its two purified fractions (LLP1 and LLP3) obtained using DEAE-52 and subsequent Sephadex G-100 column chromatography were also investigated. Antioxidant assays indicated that the polysaccharides had remarkable DPPH• and ABTS+• scavenging activities. In H2O2-induced RAW264.7 cells, LLPs showed marked protective effect on oxidative stress, reducing the MDA and ROS contents and increasing the CAT and SOD activities. LLPs were also found to effectively modulate the expressions of CAT, SOD1, Nrf-2, HO-1, NQO1 and Keap1 mRNA. Thus, the potent antioxidant effects of LLPs were achieved possibly by regulating the Keap1/Nrf-2/HO-1 signaling pathways. In lipopolysaccharide-treated RAW264.7 macrophages, LLPs reduced phagocytic ability, decreased release of NO, TNF-α, IL-1β and IL-6, and increased release of IL-10 and TGF-β1, indicating their potent anti-inflammatory effect. LLPs downregulated expressions of iNOS, IL-1β, IL-6, TNF-α, COX-2, and TLR-4 mRNA. Therefore, modulation of NF-κB signaling pathway is possibly the important anti-inflammatory mechanism of LLPs.

Bioassay‐Guided Extraction and Isolation of Natural Herbicides from Dried Zanthoxylum limonella Alston Fruits

AbstractWeeds are problematic plant species around the world. Various strategies exist for controlling weeds, but chemical treatment remains the preferred method, particularly when using natural substances. In this research, a crude aqueous‐methanol extract from dried Zanthoxylum limonella fruits was acid‐base partitioned into four fractions: neutral extract (NE), acid extract (AE), basic extract (BE), and aqueous extract (AQ). These fractions were further separated into seventeen subfractions: NEF1 to NEF7, AEF1 to AEF5, and BEF1 to BEF5, which were then tested for herbicidal activity against the growth of Chinese amaranth (Amaranthus tricolor) and barnyard grass (Echinochloa crus‐galli). Active subfractions were isolated via column chromatography and identified using spectroscopic methods, yielding seven active compounds: xanthoxyline (1), tambulin (2), atanine (3), prudomestin (4), skimmianine (5), p‐methoxybenzoic acid (6), and methyl caffeate (7). Compounds 2–7 had not been previously reported in Z. limonella. Xanthoxyline (1) was identified as the most potent botanical herbicide, fully inhibiting seed germination of Chinese amaranth and barnyard grass. This compound also decreased seed imbibition and α‐amylase activity in both species. Molecular docking studies on the α‐amylase enzyme (PDB ID: 1BG9) revealed that the aromatic, hydroxy, and carbonyl groups of xanthoxyline (1) interact with the enzyme's active sites.

Mechanosynthesis of Enaminones and Mechanochemical Telescopic Synthesis of β,γ‐Unsaturated Butyrolactams

The stereoselective mechanochemical synthesis of Z‐enaminones and bis‐enaminones was achieved without any solid component in most cases, reacting 1,3‐dicarbonyl compounds with aliphatic/aromatic primary/secondary amines, which demonstrates the viability of reacting exclusively liquid reagents. The scale‐up was robust, from five to fifty and five hundred mmol without reduction in the reaction yield. The mechanochemical telescopic synthesis of β,γ‐unsaturated butyrolactams was achieved by formal [3+2] azaanulation reaction of intermediate enaminone with maleic anhydride, boosting the access of this potentially bioactive class of compound. The mechanochemical approach revealed its sustainable face in both syntheses, avoiding solid grinding auxiliary and purification by column chromatography. A symbol was suggested to differentiate the representation of the planetary ball mill from other mechanochemical reactors, wherein three spheres are surrounded by two curved arrows representing the two inherent movements of planetary trajectories of the reaction vessel of this equipment.

Purification and Characterization of γ-Glutamyl Transpeptidase from Toona sinensisand Its Function on Formation of Sulfur-Containing Volatiles.

γ-Glutamyl transpeptidase (GGT) in Toona sinensis shoot (TS) was extracted and purified through salt precipitation, ion-exchange column, and gel filtration chromatography methods. The GGT had two peptides with M.W. 35 and 50 kDa. Its function in the formation of TS sulfur-containing volatiles (SCVs) was evaluated. The highest GGT hydrolytic activity was at pH 7.0 and 50 °C. Its activity was enhanced by Ca2+, Mg2+, K+, Na+, and Ba2+ and reduced by Cu2+, Fe3+, Mn2+, and Zn2+. The SCVs reached the highest level at GGT to TS sulfur-containing precursors extract 1:20. The aroma produced by GGT from the extract was similar to that of raw TS, especially for the cooked onion-like/TS-like flavor. These results revealed the characteristics and function of GGT in TS on sulfurous aroma formation. It could be applied to retrieve the production of sulfur-containing aroma from thermally preserved TS or similar vegetables, which lost GGT activity irreversibly during heat drying.

Synthesis of motexafin gadolinium: A promising radiosensitizer and imaging agent for cancer therapy

Abstract An improved, greener synthesis of motexafin gadolinium (MGd), a promising radiosensitizer and imaging agent that has the potential to treat or diagnose cancer, has been developed. Notably, this study addresses critical difficulties related to MGd synthesis and analysis of possible approaches, minimizing hazardous reagents/solvents, without column chromatography. The synthesis has been improved to enhance the overall yield and purity of MGd. The process changes outlined in this study are intended to lower production costs and time while maintaining the compound’s characteristics. This synthesis of MGd is an important step toward attaining its full potential in the diagnosis and treatment of cancer.

Antibacterial Activity, GC MS Analysis and In Silico Validation of Bioactive Compound from Endophytic Fungus Lasiodiplodia pseudotheobromae EF-9.

Secondary metabolites synthesized by endophytic fungi have garnered significant interest for their broad applications in treating various ailments. In this study involving 20 plant samples, 11 endophytic fungi were isolated and cultured, and Lasiodiplodia pseudotheobromae EF-9, derived from Hibiscus rosa-sinensis, demonstrated greater antibacterial efficacy than the other isolated endophytes. Phylogenetic analyses using 18S rRNA gene confirmed the EF-9 identity as L. pseudotheobromae. Following mass production, the active compound was partially purified using column chromatography. The fraction collected at the 60th min exhibited good antibacterial activity against Bacillus coagulans (MTCC 6735) and Shigella flexneri (ATCC 12022), with an inhibition zone of approximately 20 mm in diameter. UV spectral studies revealed a wide absorption band at 430 nm. High Performance Liquid Chromatography (HPLC) of the active fraction showed a distinct peak with a retention time of 4.216 min at 430 nm absorbance. Gas Chromatography-Mass Spectrometry (GC-MS) identified the active compound in the L. pseudotheobromae EF-9 culture broth extract as Bis(2-ethylhexyl) phthalate, which displayed a peak at 16.856 min and covered 66.69% of the area in the spectral analysis.

Two‐Phase Extraction, Structural Characterization of the Polysaccharide from Raspberry Pomace and Its Protection Effects against Erythrocyte Hemolysis

AbstractIn present study, a new polysaccharide (RPP) from raspberry pomace is prepared through ultrasound‐assisted aqueous two‐phase extraction (UAATPE) and purification using two‐step column chromatography, the extraction conditions are optimized by response surface method (RSM). Under the optimal parameters: liquid to material ratio of 80 mL g−1, ultrasonic power of 340 W, and ultrasonic time of 39 min, the yield of the polysaccharides is 11.72 ± 0.05%. RPP with average molecular weight (Mw) of 59 300 Da is composed of six monosaccharides. Nuclear magnetic resonance (NMR) analyses confirmed that RPP contains six main glycosidic linkages including α‐l‐Araf‐(1→, →2)‐α‐l‐Rhap‐(1→, →4)‐β‐d‐Manp‐(1→, →4)‐α‐d‐GalAp‐(1→, →3)‐β‐d‐Galp‐(1→, →4)‐β‐d‐Glcp‐(1→. RPP exhibits remarkable protective effects against H2O2‐induced erythrocyte hemolysis, the generations of reactive oxygen species, and malondialdehyde are inhibited, and the activities of glutathione peroxidase and catalase are effectively enhanced. The current findings proved that UAATPE is an effective method to extract the polysaccharide from raspberry pomace and provides a new idea for the utilization of nutrient‐rich raspberry fruit residue.

Identification of 2,4-Di-tert-Butylphenol as an Antimicrobial Agent Against Cutibacterium acnes Bacteria from Rwandan Propolis

Background/Objectives: Acne is the most prevalent dermatological condition among humans, affecting approximately 80% of adolescents during puberty. To date, numerous compounds have been used for acne treatment, including erythromycin ointments and antiseptics, with varying degrees of success. The emergence of erythromycin-resistant C. acnes strains has spurred the search for new antimicrobial agents, particularly from natural sources. Methods: Propolis collected in Rwanda was extracted and fractionated by flash chromatography and tested against C. acnes growth by using NCLSI recommendations. Results: In our research, we identified a molecule, 2,4-Di-tert-butylphenol (2,4-DTBP) which inhivbited the C. acnes growth at a concentration of 16 µg/mL. Based on these results, we formulated an ointment (1%) using OFAP18 and petroleum jelly for the potential treatment of acne using a mouse model. Conclusions: In vitro and in vivo evidence suggests that 2,4-DTBP has anti-inflammatory properties and could effectively manage the overgrowth of C. acnes as well as serve as a potent alternative for the formulation of an active propolis ointment for acne treatment.

Bacillus xiamenensis Inhibits the Growth of Moraxella osloensis by Producing Indole-3-Carboxaldehyde.

Moraxella osloensis, a gram-negative rod-shaped bacterium found on human skin, produces 4-methyl-3-hexenoic acid, contributing to clothing and body malodor. M. osloensis is resistant to UV light, drying, and antimicrobials, making its eradication challenging. As the skin is low in nutrients, commensal bacteria compete for resources and use diverse strategies to inhibit their competitors. Therefore, skin-derived bacteria that exhibited growth-inhibitory activity against M. osloensis were searched. Screening skin-derived bacteria using a coculture halo assay revealed that Bacillus xiamenensis formed an inhibition zone with M. osloensis. Coculture plates were extracted with ethyl acetate and fractionated using a silica gel column and preparative thin-layer chromatography to isolate the active compound from the B. xiamenensis metabolites. Nuclear magnetic resonance spectroscopy identified the active compound as indole-3-carboxaldehyde, which has low toxicity in humans. At soluble concentrations, indole-3-carboxaldehyde does not inhibit the growth of other bacteria, such as Staphylococcus aureus, Escherichia coli, and Bacillus subtilis, suggesting M. osloensis is highly sensitive to indole-3-carboxaldehyde. These findings highlight B. xiamenensis as a promising candidate for the development of a skin probiotic to promote skin health and combat malodor-causing bacteria.

Unlocking the Immunomodulatory Potential of Rosmarinic Acid Isolated from Punica granatum L. using Bioactivity-Guided Approach: In Silico, In Vitro, and In Vivo Approaches.

Punica granatum L. is well-known for its multifaceted therapeutic potential, including anti-inflammatory and immunomodulatory activities. This study aimed to characterize an immunomodulatory compound isolated from Punica granatum L. using a bioactivity-guided approach. Chromatographic techniques were adopted for isolation and purification of secondary metabolites. In silico, in vitro, and in vivo methods were performed to characterize the therapeutic potential of the isolated compound. Using preparative thin-layer chromatography, rosmarinic acid was isolated from F4 (column chromatography product obtained from a butanolic fraction of the extract). The impact of rosmarinic acid was assessed in rats using the neutrophil adhesion test, DTH response, and phagocytic index. In immunized rats, rosmarinic acid demonstrated significant immunomodulatory potential. Computational experiments, like molecular docking and molecular dynamics, were also conducted against two targeted receptors, Cereblon (PDB ID: 8AOQ) and human CD22 (PDB ID: 5VKM). Computational studies suggested that an increase in phagocytic index by rosmarinic acid could be attributed to inhibiting Cereblon and CD22. Pharmacokinetics and toxicity prediction also suggested the drug-likeness of rosmarinic acid. Rosmarinic acid is a potential candidate, but extensive research needs to be done to translate this molecule from bench to bedside.

Evaluation of the potential anti-arthritic effects and evaluation of acute oral toxicity of the active fraction of Eclipta prostrata leaves in rat model

The primary goal of this research was to isolate and assess the bioactive compounds within Eclipta prostrata Linn’s chloroform extract, with a focus on their potential to combat arthritis, validating its traditional use. Initially, antioxidant characteristics were evaluated, followed by fractionation through column chromatography. The in-vivo safety assessment, following OECD TG 425 Guidelines, showed no mortality at doses up to 2000 mg/kg, confirming the fraction’s safety. Subsequently, the active fraction was assessed in a complete Freund’s adjuvant-induced arthritis model. It demonstrated a dose-dependent inhibitory effect on inflammation and improved various parameters, including thermal latency time, dorsal flexion, motility, and motor coordination. Active bio constituents like rutin, stigmasterol, quercetin, and beta-sitosterol were estimated in the active fraction. The findings provide validation for the traditional use of E. prostrata as a potential plant with anti-arthritic properties, suggesting its potential suitability for rheumatoid arthritis treatment.

Chromatographic analysis of ponatinib and its impurities: method development, validation, and identification of new degradation product

BackgroundPonatinib, a third-generation tyrosine kinase inhibitor, is employed in the management of adult chronic myeloid leukemia. Nevertheless, the presence of process impurities and degradation impurities linked to ponatinib may potentially influence its effectiveness and safety. Therefore, the objective of this research was to establish a robust liquid chromatography method and systematically validate it for the detection of substances related to ponatinib.MethodsThe separation of ponatinib and its impurities was conducted using an Agilent 5HC-C18 chromatographic column (4.6 mm × 250 mm, 5 μm). The mobile phase A comprised a mixture of water and acetonitrile in a 9:1 ratio, with an aqueous solution of pH 2.4 containing 2 mM potassium dihydrogen phosphate and 0.4% triethylamine. Mobile phase B, consisting of acetonitrile, was eluted in a gradient fashion. The flow rate was set at 1.0 mL/min, detection wavelength at 250 nm, column temperature at 40°C, and injection volume at 10 μL.ResultsThe method demonstrated high specificity, sensitivity, solution stability, linearity, precision, accuracy, and robustness. Additionally, this research unveiled a novel compound, imp-B, generated via the oxidative degradation of ponatinib. The molecular structure of the newly discovered product was elucidated through the utilization of nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS).ConclusionIn conclusion, the chromatographic method developed in this study has the potential to be utilized for the detection of ponatinib and its impurities, thereby offering significant insights for quality assessment in ponatinib research.

Nostoxanthin Biosynthesis by Sphingomonas Species (COS14-R2): Isolation, Identification, and Optimization of Culture Conditions.

Nostoxanthin, a yellow pigment, belongs to the xanthophyll group of carotenoids found in various species of bacteria and cyanobacteria. Several species of Sphingomonas can produce appropriate carotenoids for survive in various environments. This comprise nostoxanthin, a significant carotenoid. The study isolated the Sphingomonas species strain COS14-R2 from the Cosmos bipinnatus and identified it through the whole-genome sequence. The strain consists of a circular chromosome with a length of 3,677,457 base pairs.s ecThe genome consists of three carotenoid biosynthesis genes, specifically crtB (phytoene synthase), crtI (phytoene desaturase), and crtY (Lycopene beta-cyclase), which are involved in the synthesis of nostoxanthin. The strain has a circular, undulated colony morphology with a deep yellow color. It demonstrates optimal growth in liquid media at 25 to 35°C and exhibits a high tolerance for pH levels between 5 and 11 and requires adequate quantities of carbon and nitrogen. We observed the highest concentration of nostoxanthin was recorded at 35°C, pH of 7.5, glucose concentration of 40g L-1, and a yeast extract concentration of 5g L-1 during dark incubation. The fed-batch fermentation process produced nostoxanthin at a concentration of 217.22 ± 9.60mg L-1, with a selectivity of 72.32% and a productivity of 2.59g/L/h. The dry biomass extract was purified using column chromatography. The LC-MS/MS analysis of the purified fraction indicated that the molecular weight of nostoxanthin is 600.5098m/z. The DPPH assay result of 75.5 ± 0.33% indicates nostoxanthin is highly effective in scavenging free radicals.

Isolation of Flavonoid Compounds Derived from Buni Leaves (Antidesma bunius (L) Spreng.)

The flavonoid compounds from the leaves of buni (A. bunius (L) Spreng.) have been isolated through a process of maceration utilizing methanol as the solvent. The concentrated methanol extract was combined with ethyl acetate. The concentrated ethyl acetate extract was subsequently dissolved in methanol and subjected to partition extraction using n-hexane. The concentrated methanol extract was acidified with 6% HCl, followed by partition extraction using chloroform. The concentrated chloroform extract was isolated using column chromatography, employing an eluent mixture of n-hexane and ethyl acetate in the following ratios: 90:10, 80:20, 70:30, and 60:40 (v/v). The compounds underwent purification via TLC preparative, resulting in a tawny paste with a mass of 30 mg and an Rf value of 0.31. The compound underwent additional identification through the application of Ultraviolet Visible (UV-Vis) spectroscopy, Fourier Transform-Infrared Spectroscopy (FT-IR), and Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), leading to the conclusion that the flavonoid in question is isoflavone.