Phytochemical Analysis最新文献

Introduction: Raw tubers of Pinellia ternata (PTRs) and processed ones (PTPs) have varied therapeutic applications in traditional medicine. PTRs are used to treat cancer, whereas PTPs are used to treat coughing and phlegm. The underlying reason for this difference is still unknown.

Objectives: This study aimed to investigate whether polysaccharides are the key differentiators in the traditional uses of PTRs and PTPs and to explore the possible mechanism for the immunomodulatory effects of polysaccharides from PTRs.

Materials and methods: PTRs and PTPs were used to produce the total polysaccharides PTR.PS and PTP.PS, respectively, which were physiochemically characterized using various techniques, such as gel permeation chromatography, nuclear magnetic resonance spectroscopy and high-performance liquid chromatography. Their immunomodulatory activity was evaluated by flow cytometry analysis in bone marrow-derived macrophages (BMDMs) and bone marrow-derived dendritic cells (BMDCs) for the analyses of activation markers and concentrations of cytokines secreted. The mechanistic study was elucidated by Western blot analysis.

Results: Physicochemical characterization revealed that PTR.PS had lower molecular weights and greater abundance of monosaccharide components, including mannose and galacturonic acid, than PTP.PS. The bioactivity analysis result showed that PTR.PS has robust immunostimulatory effects on BMDMs and BMDCs by the upregulation of NF-κB and mitogen-activated protein kinase (MAPK) pathways leading to the increased expression of activation markers and cytokine secretion. The proinflammatory M1 macrophages induce an inflammatory condition, and the activated dendritic cells can activate T cells for a cell-mediated adaptive immune response.

Conclusions: Polysaccharides are the key difference between PTRs and PTPs, resulting in different immunostimulatory effects, which are regulated through NF-κB and MAPK pathways.

Introduction: The drug "alkaloids of Sophora flavescens" (ASF) is an extract from the dried root of S. flavescens Ai. It has various pharmacological effects including anti-arrhythmia, anti-inflammatory, anti-allergic, anti-hepatitis, and antimicrobial. As there are many alkaloids with similar structure and properties, the constituent complexity brings a huge challenge in the quality control of ASF.

Objective: To develop new tandem high-performance liquid chromatography (HPLC) fingerprinting methods for the quality control of alkaloids of S. flavescens.

Methods: ASF samples were tested based on hydrophilic chromatography and ion suppression chromatographic separation mechanisms, separately. Then, the fingerprints of the two separation mechanisms were established and processed by the computer-aided tandem signal method and tandem data method, respectively. The linear quantitative profiling method was used for both qualitative and quantitative evaluation.

Results: The results of the tandem signal method and the tandem data method were accurate and consistent. The tandem signal fingerprint can reveal the inherent characteristics of traditional medicinal materials, and it is better than single fingerprints in terms of signal intensity, separation degree, homogeneity, and information abundance. The tandem data method can rapidly realize the comprehensive quality evaluation of the HPLC fingerprints under different separation mechanisms.

Conclusion: These tandem methods overcome experimental compatibility problems and the limitation of the low information content of the single separation mechanism fingerprints and can reveal the distribution and characteristics of the complex chemical components of traditional medicinal materials in a novel, realistic, and holistic way.

Background: In terms of medicinal material market and seed sources, Schisandra sphenanthera Rehd.et Wils. is often misused as Schisandra chinensis (Turcz.) Baill. Although the Chinese Pharmacopoeia makes a distinction at the medicinal material level, there is no reliable identification method for seeds. The existing approach still relies on traditional morphology, which requires rich experience.

Objectives: This study aims to fill this gap and enable quality control at the source level.

Material and methods: A neighbor-joining (NJ) tree based on the ITS2 gene was constructed to analyze species genetic relationships and find SNP site for S. chinensis. Primers targeting the SNP site were used in an allele-specific PCR system, whose sensitivity was tested with different amounts of DNA and further validated by commercial samples.

Results: The NJ tree showed that S. chinensis clustered into a single strand, clearly separated from other related species. When the primers were applied to the allele-specific PCR, a diagnostic 273 bp band was amplified specifically in S. chinensis seeds, with no cross-reactivity observed. This assay system exhibited high sensitivity, detecting as little as 0.5 ng of genomic DNA, and was validated by commercial samples, suggesting its accuracy, reproducibility, and practical applicability.

Conclusion: This method is an ideal tool for large-scale seed authentication of S. chinensis due to its high efficiency, precision, and operational simplicity. Our research contributes to quality control throughout the supply chain by ensuring the purity of the seeds at the earliest stages of cultivation, and also promotes the standardized production of this valuable medicinal resource.

Introduction: Vertigo is a prevalent clinical disorder that significantly compromises the quality of life of a patient. Among the syndrome patterns recognized in traditional Chinese medicine (TCM), vertigo due to liver-yang hyperactivity (LYH) is the most common subtype. This study explored the underlying therapeutic mechanisms of Tianma Anshen Dingxuan formula (TADF) on vertigo induced by LYH through the analysis of gut microbiota and their metabolites.

Methods: A total of 30 Sprague-Dawley rats were assigned to control, LYH vertigo model, and TADF groups. The LYH vertigo model was established through intragastric administration of aconite decoction combined with unilateral labyrinthectomy. Behavioral changes and colonic histopathology were conducted. The concentrations of 5-hydroxytryptamine (5-HT), dopamine (DA), and norepinephrine (NE) in the rat hypothalamus were quantified using the enzyme-linked immunosorbent assay (ELISA). The gut microbiota and metabolites were analyzed using 16S rRNA gene sequencing and ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC-MS). Spearman correlation analysis was performed to explore the relationships among the microbiota, metabolites, and behavioral indices.

Results: Treatment with TADF significantly improved behavioral indices (such as reduced head tilt, normalized reflexes) and colonic inflammation (lower histopathological scores, p < 0.01), whereas it significantly reduced the concentrations of 5-HT, DA, and NE in the rat hypothalamus (p < 0.05). Additionally, TADF modulated gut microbiota by upregulating both p_Patescibacteria and o_Oscillospirales while downregulating g_Bifidobacterium. Metabolomics identified 16 biomarkers (for instance, N-acetylneuraminate) enriched in pathways like tryptophan metabolism. Spearman analysis revealed correlations between microbiota (g_Ruminococcus), metabolites (chlorphenoxamine), and behavioral scores.

Conclusion: Collectively, this study is the first to demonstrate the therapeutic effect of TADF on vertigo-afflicted rats. TADF significantly improved the levels of microbiota-mediated metabolites by restoring the impaired gut microbiota.

Introduction: Physalis Calyx seu Fructus is a commonly used traditional Chinese medicine with recognized effects of clearing heat, detoxifying, soothing the throat, and resolving phlegm. Previous studies have identified it as a rich source of sucrose fatty acid esters (SEs).

Objectives: The aim of this study was to isolate structurally novel SEs with biological activity.

Material and methods: HR-LC/MS guided isolation was conducted, followed by unequivocal structural characterization of all compounds through comprehensive spectroscopic analysis. All compounds were evaluated for anti-inflammatory activity using an LPS-stimulated RAW 264.7 macrophage model, and their antibacterial activity was assessed against three clinically relevant bacterial strains. Simultaneously, isotope labeling experiments were conducted to elucidate the role of H₂O in the acyl migration process of the SEs.

Results: Ten new SEs, designated Phyalkfranose A-J (1-10), were isolated; six of them were unstable and underwent intramolecular acyl migration. Experimental results demonstrated that these 6 SEs underwent slow intramolecular acyl migration in aqueous solution, a process that was significantly accelerated under conditions of elevated temperature and high water content. Isotope labeling studies confirmed that H₂O facilitates, but does not participate in the migration. Compounds 8 and 10 inhibited the production of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α). All compounds exhibited negligible antimicrobial activity.

Conclusion: This study represents the first report on the acyl migration phenomenon of naturally occurring SEs from the plant source. These findings not only enrich the structural diversity of SEs but also offer novel strategies and methodologies for the isolation of such components.

Introduction: Venenum bufonis (VB), a traditional animal-derived medicine, exhibits significant antitumor, immunomodulatory, and analgesic activities that are primarily attributed to bufadienolides.

Objective: This study presented a novel targeted submetabolomics strategy for comprehensive profiling of bufadienolides in VB.

Material and methods: This targeted submetabolome mainly integrated scheduled diagnostic product ion (DPI) pair-based filtering and substructure recognition-based structural assembly based on the LC-MS platform. Key steps included 1) stepwise multiple ion monitoring (MIM) coupled with a dynamic exclusion function to enhance detection of minor peaks; 2) automated annotation of unconjugated bufadienolides via high-resolution extracted ion chromatograms (EICs) and a Microsoft Excel-based platform; 3) establishment of an extended tail database for conjugated bufadienolides (including dicarboxylic acid and dicarboxylic acid-amino acid conjugates) based on structural similarity and derivativity, enabling untargeted screening via validated or scheduled DPI pairs derived from MS/MS fragmentation patterns; and 4) potential conjugated bufadienolides were characterized via a substructure recognition-based strategy.

Results: The strategy identified 162 bufadienolides (39 primary, 39 secondary, and 84 tertiary) with a 96% annotation rate (155 characterized). Notably, 81 potential novel compounds were discovered, including 11 previously undescribed structural subclasses.

Conclusions: This targeted submetabolome strategy significantly expanded the diversity of bufadienolides in VB and also advanced the application of mass spectrometry in natural product discovery.

Background: Eleutherococcus senticosus leaves (ESL), recognized for their rich phytochemical profile and dual medicinal-nutritional value, hold significant potential as sustainable functional ingredients. However, the impact of post-harvest processing conditions on the retention of their bioactive components and corresponding biological activities remains to be systematically elucidated.

Purpose and study design: This study aimed to systematically evaluate the effects of different post-harvest processing methods on the quality of ESL. An integrated approach combining HPLC fingerprinting, chemometric analysis, and multi-activity evaluation (including antioxidant, antimicrobial, enzyme-inhibitory, and cellular activity assays) was employed to clarify how treatment conditions influence ingredient retention and bioactivity in ESL.

Results: The findings identified nine chemical markers in ESL, establishing a foundation for quality assessment. Moreover, the combination of vacuum freeze-drying with heated extraction (VD2) most effectively preserved key chemical markers and enhanced antioxidant capacity. This treatment also markedly promoted glutathione (GSH) biosynthesis in astrocytes under oxidative stress, showing a 77% increase compared to the control. In contrast, the VD1 treatment exhibited the strongest inhibitory activity against α-amylase and pancreatic lipase (IC₅₀ values of 0.83 ± 0.056 mg/mL and 1.61 ± 0.057 mg/mL, respectively), while the HD4 treatment showed remarkable potency in inhibiting α-glucosidase (IC₅₀ value of 0.14 ± 0.038 mg/mL).

Conclusions: This study demonstrates that specific drying and extraction protocols can selectively enhance desired bioactive properties in ESL. These findings provide a theoretical basis for optimizing industrial-scale ESL production processes, facilitating the targeted development of ESL-based nutraceuticals, neuroprotective adjuvants, and antioxidants.

Background: Pine forests are among the most extensive forest ecosystems in the world, and pine trees produce a variety of by-products, including needles, bark, seeds, and resin, which are rich in natural antioxidants and other bioactive compounds with significant potential in the nutraceutical, pharmaceutical, and cosmetic sectors. Historically, they have been undervalued and often treated as waste biomass.

Objective: This review consolidates current knowledge on the antioxidant composition of pine by-products, with a focus on their innovative applications in developing new pharmaceuticals, nutraceuticals, and cosmetic products.

Results: The chemical composition of these by-products, such as polyphenol compounds, varies based on species, geographic and environmental conditions, and extraction techniques. Emphasis is placed on green and sustainable extraction processes that preserve antioxidant potency while minimizing environmental impact. Both in vivo and in vitro studies have demonstrated a range of beneficial effects, including anti-inflammatory, anticancer, antidiabetic, and protective activities against oxidative stress-related diseases. By harnessing the antioxidant potential of pine-derived biomaterials, we can reduce waste, promote circular bioeconomy strategies, and develop innovative health-promoting products.

Conclusions: However, further research and technological advancements are needed to bring these applications to an industrial scale, serving sustainable development goals.

Introduction: Lichexanthones are the major xanthones found in lichens. They present a high degree of isomerism, which makes their identification tedious. Xanthones are known to occur in lichens according to chemosyndromes, and these compounds act as chemotaxonomic markers. Many lichens that produce xanthones are crustose lichens from Lecanora or Pertusaria genera, which often leads to small amounts of extracts being analyzed.

Objective: We aimed to set up a method able to identify the right isomers of lichexanthones contained in the extract of a xanthone-producing lichen. This method has to produce reliable results even without all the possible isomers at hand, as they are numerous and difficult to obtain.

Material and methods: The 16 norlichexanthones were obtained by a strategy combining isolation and synthesis. All of them were characterized by a full set of NMR experiments, highlighting key features, and an HPLC/DAD/MS method was developed. To exemplify the method, selected lichens were submitted to acetone microextraction and the extracts were analyzed by HPLC/DAD/MS and NMR.

Results: All norlichexanthones were well separated by HPLC/DAD/MS, which enables their identification in the lichen extracts, provided that the retention time of all the isomers is known; ¹³C NMR is very informative about the position of chlorine in norlichexanthones but lacks sensitivity. 2D NMR provides a high level of structural information even on complex extracts.

Conclusion: In addition to HPLC/DAD/MS, NMR can be used directly on a lichen extract to confirm the positions of the chlorine atoms on the lichexanthone scaffold, thanks to the HSQC experiment. Furthermore, the NOESY experiment gives the position of methylations for a comprehensive overview of the substitution patterns involved in the extract of a xanthone-producing lichen, without requiring the entire series of the 64 derivatives.

Introduction: The Olive (Olea europaea L.) is one of the most popular edible oil-producing fruits, consumed worldwide for its myriad nutritional and health benefits. Olive oil production generates huge quantities of by-products from the fruit, which are considered environmental hazards. Recently, more and more efforts have been made to valorize olive by-products as a source of low-cost, value-added food applications.

Objective: The main objective of this study was to globally assess the metabolome of olive fruit by-products, including olive mill wastewater, olive pomace, and olive seeds from fruits from two areas, Siwa and Anshas, Egypt.

Methods: Gas chromatography-mass spectrometry (GC-MS) and ultra-high-performance liquid chromatography with mass spectrometry (UPLC-MS) were used for profiling primary and secondary metabolites in olive by-products. Also, multivariate data analyses were used to assess variations between olive by-product samples.

Results: A total of 103 primary metabolites and 105 secondary metabolites were identified by GC-MS and UPLC-MS, respectively. Fatty acids amounted to a major class in the olive by-products at 53-91%, with oleic acid dominating, especially in the pomace of Siwa. Mill wastewater was discriminated from other by-products by the presence of phenolics mainly tyrosol, hydroxyl tyrosol, and α-tocopherol as analyzed by UPLC-MS indicating their potential antioxidant activity. Pomace and seeds were rich in fatty acids/esters and hydroxy fatty acids and not readily distinguishable from each other.

Conclusion: The current work discusses the metabolome profile of olive waste products for valorization purposes. Pomace and seeds were enriched in fatty acids/esters, though not readily distinguishable from each other.