Phytochemical Analysis最新文献

Introduction: Kalanchoe Adans. (Crassulaceae) is a genus of widespread succulent plants extensively studied for their biological activities. Plants of the genus are considered a potential source of antitumor agents.

Objective: This study aimed to investigate the effect of an aqueous extract and fractions of leaves of Kalanchoe fedtschenkoi R. Hamet & H. Perrier on the proliferation of melanoma cell lines employing an NMR-based biochemometric approach complemented with HPLC-DAD and UHPLC-MS/MS analyses.

Material and methods: The n-butanol fraction (KFBuOH) from K. fedtschenkoi aqueous leaf extract, which decreased B16F10 murine melanoma cells viability by 65% at 100 μg/mL, was fractionated with RP-18 SPE and Sephadex LH-20 column chromatography. The fractions were analyzed by ¹H-NMR spectroscopy and submitted to MTT cytotoxicity assays against cultured melanoma cells. Orthogonal projection to latent structures discriminant analysis (OPLS-DA) was used to correlate their ¹H-NMR profile and cytotoxic activity.

Results: This strategy enabled the identification of gallic acid (1) and two gallic acid glucosides-gallic acid 4-O-(6'-O-galloyl)-glucopyranoside) (2) and gallic acid 3-O-(6'-O-galloyl)-glucopyranoside) (3)-as putative bioactive substances, which was further corroborated by subsequent assays with enriched fractions and a gallic acid standard. The fractions enriched in gallic acid (KFA) and gallic acid galloylglucosides (KFB) evidenced selective cytotoxicity towards B16F10 cells (IC₅₀ 43.0 and 56.6 μg/mL, respectively) and MV3 human melanoma cells (IC₅₀ 93.6 and 66.1 μg/mL, respectively).

Conclusion: These results suggest a potential therapeutic use for K. fedtschenkoi in melanoma treatment. This is the first study to evidence a potential antitumor activity for gallic acid galloylglucosides.

Introduction: Drug discovery research targeting SARS-CoV-2 and other emerging pathogens remains critically important. Active compounds derived from plants frequently serve as lead compounds for further drug discovery; however, numerous unrelated chemical constituents in crude extracts may obscure the effective ingredients in LC-MS analysis.

Objective: The aim of this study is to construct a biolayer interferometry (BLI)-based system for recognizing active ingredients that inhibit the main protease (Mpro) of SARS-CoV-2 and to identify the active chemical components binding to Mpro from herbal medicines.

Methodology: We developed a novel FRET fluorogenic probe by linking the amino acid sequences of the fluorescent proteins Lssmorange and mKate2 (Ls-mK). The interaction between traditional Chinese medicine and Mpro was analyzed using BLI. Ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was employed to analyze the composition of herbal medicines.

Results: Fluorescence detection and spectroscopy confirmed the successful construction of an Mpro inhibitor screening system. Lanqin Oral Liquid (LQL) and Gardeniae fructus exhibited strong inhibitory effects on Mpro. Ten compounds were identified from G. fructus extracts; among them, deacetyl asperulosidic acid methyl ester (DAAME) and Gardoside were found to strongly bind to Mpro, with dissociation constants (KD) of 3.41 μM and 801 nM, respectively. The half-maximal inhibitory concentrations (IC50) of DAAME and Gardoside for Mpro were 27.46 and 13.7 μM, respectively.

Conclusion: This study established a functional Mpro inhibitor screening system. Among the 10 components identified from G. fructus that bind to Mpro, DAAME and Gardoside displayed strong binding and inhibitory activity, indicating their potential as lead compounds for inhibiting SARS-CoV-2 viral replication.

Introduction: Nutricosmetics derived from Silybum marianum, known for their anti-inflammatory and hepatoprotective properties, necessitate accurate quantification of silybin, a key bioactive component.

Objectives: This study aims to develop a novel high-performance liquid chromatography (HPLC) method with amperometric detection (HPLC-ECD) for the precise determination of silybin. An ultrasound-assisted extraction (UAE) procedure is also established for solid sample preparation prior to chromatographic analysis.

Materials and methods: Chromatographic separation of silybin was performed on a C18 column and using methanol-0.035 M potassium phosphate (pH 4.0) at 1.0 mL min^-1 flow rate as mobile phase in gradient mode. The electrochemical detection (ECD) of silybin was carried out on a glassy carbon electrode (GCE) at +1.10 V versus Ag/AgCl. The UAE procedure for silybin extraction from solid samples was performed by 15 min sonication in an ultrasonic bath (80 kHz and 100% power) at room temperature.

Results: Under the optimal chromatographic conditions, silybin diastereoisomers (silybin A and silybin B) can be separated from other S. marianum flavonolignans in less than 20 min, with a detection limit (LOD) of 0.060 mg L^-1 and a reproducibility (RSD) of 5%. This method was successfully applied to analyze silymarin-containing products with recoveries close to 100%.

Conclusions: This work presents the first HPLC method for silybin determination using an amperometric detector with a GCE. The LOD is competitive in comparison with previously published HPLC-DAD methods. This HPLC-ECD method allows silybin diastereoisomers identification without interferences of other flavonolignans present in silymarin extracts.

Introduction: Hypericum perforatum L. (HPL) is extensively researched domestically and internationally as a medicinal plant. However, no reports of studies related to the anticoagulant activity of HPL have been retrieved. The specific bioactive components are unknown.

Objective: The aim of this study was to develop a machine learning (ML) method for rapid prediction of anticoagulant activity in HPL and evaluation of compound activity.

Materials and methods: First, an in vitro anticoagulant activity assay was developed for the determination of the bioactivity of various medicinal parts of HPL. Then, the peak areas of compounds in HPL were integrated using UPLC-Q-TOF-MS analysis. Subsequently, nine independent ML methods and two ensemble learning methods have been established to predict the anticoagulant activity of HPL and to evaluate the contribution of compounds. Feature importance scores were used for models visualization.

Results: A total of 24 compounds were shown to exhibited superior anticoagulant activity. Molecular docking experiments likewise confirmed this result. The results show that the branches of HPL have excellent anticoagulant activity, which has been previously overlooked. The established ML model demonstrated good performance in the prediction of the activity of HPL.

Conclusion: The results were accurate and reliable, which significantly improved the efficiency of active compounds screening, and further exploration in this area is warranted.

Introduction: Targeted screening of inhibitors of key enzymes in the progression of diabetes from natural products is one of the effective methods for the treatment of diabetes. Polygala has been proved to reduce glucose levels; however, the bioactive compounds in Polygalae Radix (PR) that have anti-diabetic properties are unknown.

Objective: The purpose of this study was to explore the material basis of the anti-diabetic effect of PR by inhibiting α-amylase through an online detection system and molecular docking.

Methods: An online analysis platform was established and optimized for the screening of potent enzyme inhibitors from complex mixtures based on ultra-performance liquid chromatography-photodiode array-quadrupole-time-of-flight-mass spectrometry-α-amylase-fluorescence detector (UHPLC-PDA-Q-TOF-MSⁿ-α-amylase-FLD) detection system and molecular docking, which could efficiently separate extracts, quickly detect α-amylase inhibitors, and determine their structures. Molecular docking confirms the inhibition of these compounds. The molecular interaction between α-amylase and the active compound was evaluated.

Results: Among the 101 compounds identified, 28 compounds had a strong inhibitory effect on α-amylase. Molecular docking screening confirmed the inhibition of these compounds and evaluated the molecular interactions between α-amylase and 30 active compounds, which strongly supported the experimental results. Among the evaluated compounds, onjisaponin R (83) and polygalaxanthone III (11) have the strongest inhibitory activity to α-amylase (the binding energies were -9.639 and -8.972 kcal/mol, respectively) and are potential lead compounds against diabetes.

Conclusion: This study proved the feasibility of using the existing platform to screen the active ingredients in PR extract, and provided a practical method for the rapid screening of potential anti-diabetic active ingredients in traditional Chinese medicine.

Background: Ginseng volatile oil (GVO) is a valuable active ingredient in ginseng (Panax ginseng C. A. Mey.) with high research potential. Drying procedures alter the real composition of the fresh material, for example, the evaporation of compounds with low boiling point. In this study, the composition of volatile oil in fresh ginseng (FG), sun-dried ginseng (SDG), and red ginseng (RD) was systematically analyzed to clarify the dominant components of FG and their potential pharmacological effects, which provides a basis for application and development of FG.

Methodology: GVO was obtained through water vapor distillation and analyzed using GC-MS. Pattern recognition analysis was employed to differentiate components in three processed types of ginseng. Based on this analysis, the active ingredients and key targets were screened. The binding mode and affinity were verified using molecular docking technology. Finally, the anticancer activity of GVO was verified by cell experiments.

Results: A total of 53 components were identified in three processed types of ginseng by GC-MS. Among them, 32 differential components were screened by pattern recognition analysis. Ultimately, 6 active ingredients (panaxydol, nerolidyl acetate, falcarinol, cis-β-farnesene, γ-elemene, and β-elemene) and 15 key targets were determined by network pharmacology analysis. Molecular docking results revealed that β-elemene exhibited a higher affinity with EGFR, ESR1, and ERK2. Cell experiments indicated that GVO promotes apoptosis in cancer cells.

Conclusion: This research proposed a strategy that integrated "component detection-virtual multitarget screening-active component prediction-experimental verification" to expedite the identification of active ingredients, providing insights for application of FG and the development of functional products.

Introduction: This study evaluates the effectiveness of Traditional Chinese Medicine (TCM) extracts in blocking the interaction between the SARS-CoV-2 Spike protein and human ACE2 receptor, utilizing a dual-method approach to explore the antiviral potential of natural compounds.

Objectives: This work aims to evaluate the capability of TCM extracts in inhibiting the SARS-CoV-2 Spike protein and ACE2 receptor interaction using advanced biochemical assays.

Methods: A dual-method screening approach was utilized, beginning with a pseudovirus assay to assess the inhibition capabilities of TCM extracts in vitro, followed by a split-ubiquitin yeast two-hybrid (Y2H) system to validate interactions in live cells. Active compounds were characterized and quantified using UPLC-Q-Exactive-Orbitrap-MS.

Results: Among the 91 TCM extracts tested, Rosmarinus officinalis exhibited the most potent inhibition in both pseudovirus and Y2H assays, significantly reducing viral entry and disrupting the Spike-ACE2 interaction. Comprehensive chemical profiling via UPLC-Q-Exactive-Orbitrap-MS identified 132 compounds, including phenolics, flavonoids, and terpenoids.

Conclusion: This research validates the use of TCM extracts in viral inhibition strategies, demonstrating the utility of integrating traditional remedies with modern scientific approaches to discover new therapeutic agents.

Introduction: This study explores the cytotoxic activity-guided isolation of the underground parts of Laserpitium hispidum M. Bieb and Laserpitium petrophilum Boiss. & Heldr., which have not been previously investigated.

Objectives: The aim is to isolate and evaluate bioactive compounds from Laserpitium L. species with anticancer potential.

Material and methods: This study involves bioactivity-guided isolation and structural studies of the pure compounds utilizing NMR, UV-Vis, IR spectroscopies, and HRMS. The cytotoxic activity of the isolated compounds was evaluated in vitro and in vivo, whereas molecular modeling, docking, and ADME predictions were conducted using Schrödinger software.

Results: The study isolated phenylpropanoids (laserine (1), latifolone (2), myristicin (3)), sterol (stigmasterol (4)), polyenes (falcarindiol (5)), sesquiterpene lactone (11-hydroxybadkhyzin (6)), and nordaucane sesquiterpene (norlasidiol angelate (7)) from L. hispidum, whereas L. petrophilum yielded 10β-acetoxy-8α-angeloyloxy-6αH,7αH-guaian-3-en-12,6-olide (8), 10β-acetoxy-8α-senecioyloxy-6αH,7αH-guaian-3-en-6,12-olide (9) and acetylisomontanolide (10). Molecular docking simulations revealed stable interactions between compounds 7 and 9 with estrogen receptor α (ERα) and vascular endothelial growth factor receptor 2 (VEGFR2), with compound 7 showing superior stability and binding affinity. In silico ADME predictions indicated favorable pharmacokinetic properties, including high oral absorption.

Conclusion: Compounds 7 and 9, representing new nordaucane and sesquiterpene lactones, have not been previously reported. In vitro cytotoxicity revealed that compound 7 exhibits potent anti-cancer activity against MCF-7 cells, whereas compound 9 showed reduced cytotoxicity. In vivo testing in Caenorhabditis elegans supported these findings, suggesting safety and efficacy in organisms. In silico results emphasize the potential of these compounds, with compound 7 promising due to its stability and strong binding affinity.

Introduction: This study investigates the composition and antibacterial properties of essential oil extracted from Pinus koraiensis (Siebold & Zucc) pine needles using a liquid nitrogen freezing treatment combined with solvent-free microwave extraction (LNSFM).

Objective: The aim is to develop a low-energy, high-efficiency extraction method for conifer essential oils, analyze their chemical composition, and evaluate their antibacterial efficacy.

Methodology: Pine needle samples were frozen with liquid nitrogen and subsequently crushed. The essential oil was extracted using solvent-free microwave technology. A single-factor test and response surface methodology were employed to optimize extraction parameters. The extraction efficiency of LNSFM was compared with traditional methods through kinetics, and the essential oil components were analyzed using gas chromatography-mass spectrometry (GC-MS). The antibacterial activity of the extracted volatile oil was tested against Escherichia coli and Staphylococcus aureus.

Conclusion: LNSFM proves to be a green and efficient extraction method suitable for obtaining volatile oils from pine needles, which demonstrate significant antibacterial properties.