This study developed a rapid and green liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of three major flavonoids (cianidanol, saxifragin, and miquelianin) in alpine bistort rhizome (ABR) and Zhuyaliao Zhixie Granules (ZZG). To our knowledge, this is the first simultaneous determination of these three compounds in Polygonaceae. Following ultrasonic extraction with 70% methanol, separation was achieved on a Poroshell 120 EC-C18 column using 0.1% formic acid in water and methanol as the mobile phase. LC-MS/MS detection was performed in the negative ion mode using multiple-reaction monitoring, with a total analysis time of only 6 min per sample. All analytes showed excellent linearity (r > 0.998) and satisfactory recoveries (94.0-107%) with low relative standard deviations (1.06-2.87%). Quantitative profiling revealed that both source type and provenance significantly affected flavonoid levels in ABR, whereas the granules exhibited markedly reduced levels of these flavonoids compared to the raw botanical materials. Compared to previously reported LC methods, this approach exhibited greater analytical efficiency and was both practical and green according to the Blue Applicability Grade Index (BAGI). The developed method provides a reliable tool for ABR quality evaluation and promotes greener, faster modernization of traditional Chinese medicine analysis.

This study developed a rapid and eco-friendly HPLC method for simultaneous determination of adenosine and ergothioneine in Cordyceps spp. samples. The extracted protocol involved dispersing samples in diatomaceous earth followed by vortex extraction with 80 % methanol. Chromatographic separation was achieved within 5 min using a Shim-pack Velox HILIC column with a gradient mobile phase of 0.1 % formic acid aqueous solution and acetonitrile solution containing 0.1 % formic acid and detection at 260 nm. Both analytes demonstrated excellent linearity (r > 0.9999) with average recoveries of 102.82 % (adenosine) and 100.10 % (ergothioneine), accompanied by RSDs below 2 %. The contents of adenosine and ergothioneine in 46 batches of Cordyceps spp. samples were 0.000-2.700 mg/g dry weight (DW) and 0.000-0.703 mg/g DW, respectively. The Ophiocordyceps sinensis and its related species were successfully classified by hierarchical cluster analysis. This study determined the content of ergothioneine in Cordyceps spp. samples for the first time, and identifies ergothioneine as a critical marker for differentiating authenticated Ophiocordyceps sinensis from its adulterants. Results of BAGI assessment (score=87.5) confirmed the superior practicality and environmental friendliness of the developed method. This rapid, accurate, and reliable method provided a scientific foundation for enhancing quality evaluation technology of Cordyceps spp. species samples.

ETHNOPHARMACOLOGICAL RELEVANCE:Citri Grandis Exocarpium (CGE), a traditional Chinese medicinal herb with a long history of medicine and food homology, has been documented in Gleanings from the Materia Medica for its efficacy. Modern research has revealed that the active components exhibit anti-inflammatory properties and metabolic regulatory functions, emerging as a promising candidate for the treatment of obesity. AIM OF THE REVIEW:This review aims to conduct a comprehensive analysis of the pharmacological material, mechanisms, and clinical translational prospects underlying the anti-obesity effect of CGE. It also proposes to evaluate its potential utility as a medicine and food homology in obesity prevention and treatment, and address its limitations, and propose solutions for existing research in this field. METHODS:This review primarily focused on elucidating the mechanism and effectiveness of CGE in treating obesity and its related complications, by searching across electronic databases, including PubMed, Web of Science, Scopus, Google Scholar, and CNKI. RESULTS:The effective phytochemicals, molecular mechanisms, and product applications of CGE in the prevention and treatment of obesity and related complications were summarized. Specifically, naringin, limonene, and other key components are crucial in activating the AMPK signaling pathway to inhibit lipid accumulation, and regulating the gut microbiota to improve metabolic inflammation. In addition, CGE exerts regulatory effects on lipogenesis and energy metabolism by influencing crucial genes such as Ucp1 and Pgc-1α, positioning CGE as a potential therapeutic drug and healthy food for addressing obesity and its associated metabolic disorders. CONCLUSION:Owing to the synergistic effect of its diverse bioactive constitutes as well as its medicinal-food application properties, CGE has become a promising candidate for anti-obesity interventions. However, existing studies have certain limitations that have not been widely and deeply investigated. In the future, more rigorous clinical trials, innovative delivery technologies, and internationally recognized standards are required to support the transition of CGE from traditional empirical practice to an evidence-based therapeutic option.

Taihangia rupestris Yu & Li (T. rupestris) was a nationally protected plant and its active compounds exhibit antidiabetic potential, but systematic phytochemical and pharmacological investigations remain limited. To balance conservation and utilization, the chemical composition and bioactivities (antioxidant and α-glucosidase inhibitory effects) of wild and cultivated T. rupestris were compared, followed by active compound screening. UPLC-MS/MS was used to identify chemical constituents in wild, mountain-cultivated, and foothill-cultivated samples, with multivariate analysis for differential components. The TFC and TPC, and the antioxidant capacity were evaluated via FRAP, CUPRAC, TRC, and DPPH assays. Additionally, α-glucosidase inhibition was assessed via IC50 determination. Online HPLC-ABTS and ultrafiltration-LC/MS (UF-LC/MS) were employed to screen antioxidants and α-glucosidase inhibitors (α-GIs), respectively, with molecular docking validating binding mechanisms. As a result, among 114 identified compounds, 111 showed significant environment-dependent variations, primarily flavonoids, phenolics, and terpenoids. Foothill-cultivated plants exhibited upregulated flavonoids/phenolics (e.g., rutin and gallic acid derivatives, P < 0.05) and superior antioxidant activity (Trolox equivalents: FRAP 367.18 ± 1.03; CUPRAC 572.40 ± 0.82) and α-glucosidase inhibition (IC50 0.2775 µg mL-1) versus wild (IC50 0.4948 µg mL-1) and mountain-cultivated samples (IC50 0.5425 µg mL-1). Ten antioxidants were screened, with seven also acting as α-GIs. UF-LC/MS and docking confirmed 8 α-GIs (binding energy < -5 kcal mol-1), where phenolic hydroxyl groups formed hydrogen bonds with ASP residues of α-glucosidase. Cultivated T. rupestris (especially foothill-grown) outperformed wild plants in bioactives composition and efficacy, serving as a sustainable alternative. Flavonoids and phenolics contributed to its antidiabetic potential via dual antioxidant and enzyme-inhibitory effects, supporting further pharmaceutical development.

Bistorta vivipara L. achenes (B. vivipara achenes), rich in phenolic acids and flavonoids, are a valuable food and medicinal resource. However, a detailed analysis of its individual components and their mechanisms remains unexplored. This study introduces solid-phase extraction (SPE) for selectively isolating and purifying total phenolic acids and flavonoids from B. vivipara achenes. The antioxidant potential of the extracts was evaluated using ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), DPPH (2,2-diphenyl-1-picrylhydrazyl), and FRAP (ferric ion reducing antioxidant potential) assays. Results showed significant antioxidant activity in the total phenolic acid extract (125.1 ± 0.7 mg GAE/g dry weight [DW]) and total flavonoid extract (1491.1 ± 19.9 mg RE/g DW). Using online FRAP–high-performance liquid chromatography (FRAP–HPLC), seven antioxidant components were identified in the phenolic acid fraction and thirteen in the flavonoid fraction. Notably, protocatechuic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, shaftoside, isoshaftoside, isoorientin, taxifolin, and vitexin are reported for the first time as antioxidant constituents in B. vivipara achenes. These findings highlight B. vivipara achenes as a promising source of natural antioxidants and functional food ingredients. The SPE–FRAP–HPLC method enables comprehensive analysis by selectively enriching active compounds and enhancing sensitivity and accuracy compared with crude extracts. Furthermore, the application of online FRAP–HPLC facilitates real-time monitoring of the antioxidant activity of individual compounds during chromatographic separation, providing a detailed antioxidant activity profile of phenolic compounds in B. vivipara achenes.

Cordyceps (Chinese caterpillar fungus) is a precious medicinal and edible fungus-larva complex that is naturally formed in a specific environment. However, limited data have been reported on spatial alterations in the lipidome and metabolome of larvae before and after entomopathogenic fungal infection. In this work, changes in chemical compositions and their spatial distribution patterns between host larva and the fungus-larva complex were visualized and characterized using matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI). Liquid chromatography-mass spectrometry (LC-MS) was performed to support MALDI MSI results. Our work revealed the single dominant distribution of many compounds in the host larva, as well as stroma and sclerotium of Cordyceps. Based on further analysis, the heterogeneous distribution of certain lipid classes across the host larva, stroma and sclerotium of Cordyceps was revealed, reflecting key lipid metabolic events, such as assimilation, dissimilation, and synthesis of derivatives. Small metabolites were also detected, and an increase in both the amount and the kinds was observed from larvae to Cordyceps.

As a rare and precious traditional Chinese medicine (TCM), Ophiocordyceps sinensis (O. sinensis) is plagued by various adulterants in the market due to factors such as resource scarcity, which seriously affects medication safety and clinical efficacy. Existing identification methods generally have problems including strong subjectivity, high sample destructiveness, long detection cycle, and complex operation, and it is difficult to identify different types of adulterants using the same method. Innovatively, this study adopted scanning electron microscopy-energy dispersive x-ray spectroscopy (SEM-EDS) technology to establish a rapid, accurate, and widely applicable method for identifying adulterated O. sinensis. Authentic O. sinensis samples from major producing areas such as Xizang, Qinghai, Sichuan, and Gansu, as well as different types of adulterated samples, were collected. Using SEM-EDS technology, combined with micromorphology and elemental composition, the differences between authentic and adulterated samples were systematically revealed through indicators including morphological differences, mass fraction of characteristic elements, and surface area coverage (SAC) of characteristic elements in surface scanning. This study confirmed that SEM-EDS technology could simultaneously obtain the micromorphological and elemental distribution information of samples, realizing rapid and accurate identification of multiple types of adulterated O. sinensis. The proposed method provided new technical support for the quality control and market supervision of O. sinensis, which could be extended to the adulteration inspection of other precious and delicate TCM.

Cultured Cordyceps sinensis (CCS) improves sperm quality in patients with oligoasthenozoospermia (OAS), but its mechanisms remain unclear. A cyclophosphamide (CTX)-induced OAS mice model was employed to evaluate the effects of CCS on hormone secretion, blood-testis barrier (BTB) integrity, testicular oxidative stress, and spermatogenic cell apoptosis. The steroidogenic factor-1 (SF-1)-mediated steroid hormone biosynthesis was further examined. Our results demonstrated that CCS elevated serum hormone levels, increased BTB-related protein expression, and inhibited spermatogenic cell apoptosis. Furthermore, CCS upregulated the protein expression of steroidogenic enzymes and SF-1 in testes. In vitro, CCS extract enhanced testosterone production and steroidogenic enzyme expression in CTX-injured TM3 cells, concomitant with the upregulation of SF-1. Moreover, SF-1 knockdown reversed the effect of CCS extract on testosterone production. In conclusion, CCS improves OAS by enhancing testosterone production primarily via activation of SF-1-mediated steroid hormone biosynthesis. The increased testosterone contributes to maintaining BTB integrity and inhibiting testicular cell apoptosis.

A rapid and environmentally friendly analytical method was developed and validated for the simultaneous quantification of four antibiotic residues (enrofloxacin, ofloxacin, sulfamethizole, and trimethoprim) in bear bile powder. The optimized procedure features a simplified sample preparation with minimal solvent consumption and a fast chromatographic separation. Method validation confirmed excellent performance in specificity, linearity, precision, accuracy, and stability. Furthermore, the method’s greenness and practicality were quantitatively superior to a conventional approach, as evidenced by higher scores in sustainability assessments. Application to market samples revealed a notable disparity: Antibiotic residues were frequently detected in bear bile powder, while none were found in biotransformed bear bile powder. These findings furnish a reliable analytical tool for quality control and highlight biotransformed bear bile powder as a safer alternative, underscoring the impact of production practices on product safety.

Ophiocordyceps sinensis (Berk.) is a functional food with health. O. sinensis quality varies by geographical origins, and current identification methods are sophisticated and time-consuming. This study aims to develop a rapid and straightforward method for accurately identifying O. sinensis geographic origins. Surface-enhanced Raman spectroscopy (SERS) was applied to analyze O. sinensis from four major production areas in China. Liquid chromatography-mass spectrometry (LC-MS) was used as a reference method to characterize compositional differences among samples and to verify the geographical authenticity of O. sinensis from the four production areas. Six machine learning (ML) algorithms were introduced for predicting geographical origins, and evaluation metrics were used to assess model performance. According to the comparative analysis, the Support Vector Machine (SVM) model performed best with the highest discrimination accuracy. A feature importance map was constructed to understand further how the model makes predictive decisions, revealing the significant Raman shifts in classifying O. sinensis from different geographical origins. The SERS-SVM method developed in this study contributes to the authenticity identification of O. sinensis geographical origins. It shows the potential to serve as an effective quality control method for the valuable TCM (traditional Chinese medicine).

Ophiocordyceps sinensis is a premier medicinal fungus with a unique slow-killing parasitic strategy, yet the molecular basis of its long-term persistence in host hemocoel remains elusive. This fungus coexists with Thitarodes xiaojinensis larvae for over six months, ultimately leading to host death at the last instar before pupation. To decode the fungal survival strategies masked by host resistance, this study integrated omics approach combining sublethal imidacloprid-induced transcriptomics with longitudinal metabolomic profiling. We investigated the transcriptomic response of O. sinensis within the larval hemocoel under exposure to a sublethal dose of imidacloprid, a stressor utilized here to suppress host immunity via neuro-immune crosstalk. This approach allowed us to infer the core molecular responses of O. sinensis across three critical infection stages (30, 90, and 150 days post-infection, dpi). Differential gene expression and functional enrichment analyses revealed that O. sinensis significantly responds to host immune disturbance induced by sublethal dose of imidacloprid. During the early infection stage (30 dpi), Imidacloprid exposure suppressed fungal cell proliferation genes while activating secondary metabolite pathways and immune defense genes. This suggests that during a conventional infection, O. sinensis secondary metabolism was suppressed under the host immunity to maintain a balanced blastospore proliferation. Beyond similar response in fungal secondary metabolites and host immunity, imidacloprid also suppressed protein biosynthesis during the mid-infection stage (90 dpi), indicating that the fungus undergoes metabolism suppression while continuing to maintain balance between blastospore proliferation and host immunogenicity for the conventional infection. During the late stage (150 dpi), genes associated with hydrolytic activity, transport and cell wall degradation were upregulated while fatty acid synthesis genes were downregulated by imidacloprid, suggested that the conventional infection of O. sinensis at this stage exhibits metabolic polarization to shift blastospore to hypha, tightly regulating hydrolysis to balance nutrient acquisition with host viability and increasing fatty acid biosynthesis. Metabolomic analysis showed that the accumulation of chitin precursors in the late stage, including D-glucosamine-6-phosphate, marks the critical blastospore-to-hyphae transition. Furthermore, the accumulation of compounds like chlorogenic acid, L-glutathione oxidized, and hesperidin indicates the dual responses of secreting antimicrobials and deploying antioxidants to protect and sanitize the nutrient base. This study provides insights into the dynamic, stage-specific gene expression profiles of O. sinensis that enable its long-term survival in the host hemocoel. Through coordinated regulation of proliferation, metabolism, secondary metabolite production, and immune evasion, the fungus sustains persistent infection and successfully completes its life cycle. These findings advance our understanding of the strategies employed by slow-killing fungal parasites.

Ophiocordyceps (Ophiocordyceps sinensis) is a precious medicinal and edible fungus widely utilized in nutraceuticals and functional foods. However, the structural intricacies and biological potential of its high-molecular-weight polysaccharides remain largely unknown. In the present study, a novel high-molecular-weight neutral polysaccharide, designated as CSP1b, was isolated and purified from cultured ophiocordyceps via water extraction, ethanol precipitation, and sequential column chromatography. Characterization revealed that CSP1b possesses an extraordinary molecular weight of 2.65 × 107 Da and consists predominantly of glucose (73.98%), mannose (13.35%), and galactose (12.67%). Structural elucidation identified CSP1b as a galactomannoglucan with a backbone of →4)-α-D-Glcp-(1 → 6)-α-D-Glcp-(1→, branched at the O-6 position by side chains containing →6)-α-D-Glcp-(1→, α-D-Manp-(1→, and various galactose residues, resulting in a branching degree of 32.06%. The polysaccharide exhibited a filamentous and ribbon-like intertwined surface morphology. In vitro immunomodulatory assays demonstrated that CSP1b significantly enhanced the proliferation and phagocytic capacity of RAW 264.7 (mouse macrophage cell line), as well as induced the secretion of nitric oxide (NO) and cytokines (TNF-α, IL-6) in a dose-dependent manner. In summary, a novel high-molecular-weight galactomannoglucan (CSP1b) was isolated for the first time from cultured ophiocordyceps and exhibited potent immunostimulatory activity. These findings enrich the polysaccharide library of ophiocordyceps and support its application in immunoregulatory products.

Pulmonary fibrosis (PF) is a progressive lung disease driven by inflammation and oxidative stress, with fewer effective therapeutic options. In this study, we investigated Cordyceps sinensis, a traditional edible mushroom, as a potential dietary solution. The ethanolic extract of C. sinensis (CF0), identified as the most biologically active fraction, was subsequently subjected to UPLC-Q-TOF-MS analysis. CF0 revealed a chemically diverse profile enriched in polyphenolic constituents, nucleoside analogs (including cordycepin and adenosine derivatives), sterols, sphingoid bases, and low-molecular-weight compatible solutes. We demonstrate that CF0 significantly inhibits TGF-β1-induced myofibroblast activation in vitro and preserves lung architecture in bleomycin-challenged mice in vivo. At the molecular level, the extract worked two ways: it dampened the inflammatory “fire” (the TLR4/MyD88/NF-κB pathway) and boosted the body’s natural antioxidant defense (the Nrf2/HO-1 pathway), thereby protecting lung DNA from damage. Additionally, CF0 restructured the gut microbiota (Akkermansia, Lactobacillus) and restored short-chain fatty acids (SCFAs). This study proposes a chemistry-biology alignment supporting a redox-centric mode of action: polyphenolic motifs and cordycepin directly engage the Keap1-Nrf2 pathway, while gut-derived SCFAs systematically inhibit NF-κB, reinforcing the gut–lung barrier. Collectively, these findings establish C. sinensis as a promising functional food that delivers multi-target anti-fibrotic protection by coordinating redox homeostasis with gut–lung axis regulation.

This study presents a rapid and eco-friendly analytical method for quantifying hesperidin and naringin in mandarin peel, orange peel, and pomelo peel, thereby promoting resource utilization and sustainable practices. The method employed straightforward grinding with diatomite for 2 min, followed by extraction using a mixture of N,N-dimethylformamide and methanol (40:60, v/v) with ultrasonic assistance for 2 min, which facilitates the efficient release of bioactive flavonoids. The analysis was performed using high-performance liquid chromatography with a Agilent InfinityLab Poroshell 120 EC-C18 column at 30 °C, employing a gradient elution of 0.1

Mandarin peel and leaf were investigated as candidate ingredients for the development of functional foods and feeds. A total of 37 compounds were identified; among them, 2 and 14 were detected for the first time in peel and leaf, respectively. Leaf exhibited richer chemical diversity, whereas peel showed higher contents of the key bioactive compounds (hesperidin, nobiletin, and tangeretin). The contents of these three compounds varied significantly among peel samples from different cultivars. Extraction dynamics studies indicated that, for daily consumption, three cycles of hot-water infusion or approximately 24 h of liquor steeping are the optimal conditions. Network pharmacology and molecular docking analyses suggested that hesperidin, nobiletin, and tangeretin may exert beneficial effects in ameliorating cancer and cardiovascular diseases. Overall, these findings highlight the health-promoting potential of mandarin byproducts and support sustainable development through their valorization into functional ingredients.

Magnolol and honokiol, the primary active components in Magnolia Officinalis Cortex (Houpo), are key indicators for quality control. However, existing analytical methods suffer from prolonged analysis cycles, excessive consumption of hazardous solvents, and high costs, which hinder their application in large-scale, high-throughput, and eco-friendly quality assessment. In this study, an innovative, rapid, and green analytical strategy was developed through the integration of matrix solid-phase dispersion (MSPD) extraction with dual-wavelength ultraviolet spectrophotometry (DUV). This method achieves integrated efficient extraction and purification of Houpo samples, facilitating rapid detection without a mobile phase. Compared with the Chinese Pharmacopoeia (ChP) reference method (>24 h), the proposed method reduced the total analysis time to 3 min, while only 5.4 mL of ethanol-a green solvent-was utilized. The reliability of the method was fully validated in strict accordance with ICH Q2(R1) guidelines and the ChP, showing no significant differences compared with the ChP method (p > 0.05). Furthermore, the AGREEprep score of this method reached 0.72, which was superior to other previously reported methodologies. Meanwhile, the White Analytical Chemistry (WAC) evaluation also confirmed the excellent comprehensive performance of the proposed method. This technique provides an efficient, accurate, and environmentally sustainable solution for the quality control of Houpo, enables high-throughput screening in grassroots laboratories and industrial production sites, and further promotes the modernization of quality evaluation systems for traditional Chinese medicine.

A comprehensive evaluation was conducted on the bioactive compounds and antioxidant capacities in different plant parts (achene, cauline leaf, and rhizome) of alpine bistort, Bistorta vivipara (L.) Gray. Qualitative analysis identified 76 compounds, with flavonoids and organic acids as the predominant chemical classes across all plant parts. Quantitative analyses indicated variations in the content of these compounds across plant parts. Cultured alpine bistort exhibited a consistent trend of higher phytochemical content and greater uniformity compared to the natural alpine bistort across all plant parts. Four compounds (cianidanol, miquelianin, saxifragin, and neochlorogenic acid) showed potential as part-specific discriminators. Among all plant parts evaluated, the rhizome demonstrated the highest antioxidant capacity. Network pharmacology and molecular docking analyses indicated that flavonoids and organic acids serve as the primary antioxidant compounds, acting through multiple targets and pathways. These findings provide important evidence supporting the industrial application of alpine bistort.

Cultivated Chinese cordyceps (CCC), different from the cultivated Cordyceps sinensis mycelia, is regarded as the ideal substitution for wild Chinese cordyceps. As the main active component in Chinese cordyceps, polysaccharide quantification is important for its quality assessment. In this study, a major polysaccharide (CSWP-40) of CCC, characterized as a glucan with immune enhancing activities, was purified as the standard reference. A high-performance size exclusion chromatography method was developed and validated for the polysaccharide quantification. The results of different grade CCC tests revealed that CSWP-40 content in each strand of the big CCC (0.45-0.55 g/strand) was 6 times higher than that in small CCC (0.18-0.22 g/strand). Through meticulous source tracing, it was discovered that CSWP-40 was predominantly derived from the host caterpillar. Additionally, CSWP-40 can distinguish Chinese cordyceps from four common adulterants (Cordyceps hawkesii, Cordyceps liangshanensis, Cordyceps gunnii, and Cordyceps taii), which lack detectable CSWP-40. It is worth mentioning that CSWP-40 was also detected in Cordyceps gracilis and Cordyceps militaris, which implies that it is not exclusive to Chinese cordyceps. In summary, this study demonstrates that CSWP-40, a host caterpillarderived immunoactive polysaccharide, can be established as a central biomarker for quality assessment of CCC. Quantitative analysis of CSWP-40 not only facilitates accurate authentication and product-grade optimization but also provides critical quality control parameters for standardized manufacturing of high-value-added Cordyceps-derived products (e.g., pharmaceuticals and nutraceuticals), thereby enhancing industrial standardization and market credibility.