Through several chromatographic methods, such as silica gel, Sephadex LH-20, medium-pressure liquid chromatography (MPLC), combined with modern spectroscopic techniques including nuclear magnetic resonance (1D and 2D NMR) and high resolution mass spectrometry (HRMS), two iridoids and six oleanane-type triterpenoids were isolated and identified from the roots and rhizomes of Patrinia scabiosaefolia. Among them, compound 1 was a new 3, 8-epoxy iridoid and named as patrinin C (1). Compounds 2-8 were isolated from this plant for the first time, and their structures were identified as alboside II (2), 11α, 12α-epoxy-3β, 23-dihydroxyolean-28, 13β-olide (3), 23-hydroxy-3-oxo-28-nor-12, 16-dien oleanane (4), 3′-O-acetyl-3-O-α-L-arabinosyl-23-hydroxyolean-12-en-28-oic acid (5), 2′, 3′-O-diacetyl-3-O-α-L-arabinopyranosyl-23-hydroxyolea-12-en-28-oic acid (6), hederagenin 3-O-(2-O-acetyl-β-D-xylopyranosyl)-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside (7) and pictoside A (8), respectively.

By comprehensively applying various separation techniques such as silica gel, ODS, and Sephadex LH-20 column chromatography, as well as preparative HPLC, fourteen C21 steroidal compounds were isolated from the chloroform extract of Cynanchum wallichii Wight. The structures of these compounds were identified through spectroscopic techniques including mass spectrometry (MS), nuclear magnetic resonance spectroscopy (NMR), ultraviolet spectroscopy (UV), and infrared spectroscopy (IR) methods as cynanotonin E (1), qingyangshengenin (2), qinyangshengenin-3-O-β-D-cymaropyranoside-(1→4)-β-D-cymaropyranoside (3), qinyangshengenin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosy-(1→4)-β-D-digitoxopyranoside (4), qinyangshengenin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside (5), 12-O-benzoylsarcostin-3-O-β-D-oleandropyranosy-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranosyl (6), 12-O-benzoylsarcostin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranosyl (7), caudatin-3-O-β-D-digitoxopyranoside (8), caudatin-3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranoside (9), caudatin-3-O-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside (10), caudatin-3-O-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside (11), otophylloside B (12), otophylloside E (13) and otophylloside F (14). Among them, compound 1 is a new compound. The cytotoxic activities of compounds 1-14 against human hepatocellular carcinoma HepG2 cells were tested using the MTS method. The results indicated that all compounds exhibited good inhibitory activities against the proliferation of HepG2 cells, with IC₅₀ values ranging from 4.3 ± 1.25 - 26.3 ± 1.65 μmol·L^-1. Among them, the inhibitory activities of compounds 1, 3 and 6-14 were all stronger than that of the positive control cisplatin (DDP, IC₅₀: 15.9 ± 1.25 μmol·L^-1).

The chemical constituents in the bark of Eucommia ulmoides were isolated and purified using chromatographic techniques, including silica gel, Sephadex LH-20, ODS, MCI, and semi-preparative HPLC. The structures of the compounds were elucidated by spectroscopic analyses (NMR, IR, UV, and MS). A total of six compounds were identified as duzhonglactone (1), (7S, 8R)-4-hydroxy-3, 3′, 5′-trimethoxy-8, 9-dinor-8, 4′-oxyneoligna-7, 9-diol-7′-aldehyde (2), armaosigenin (3), (-) evofolin B (4), N-trans-feruloyltyramine (5), and C-veratroylglycol (6). Among them, compound 1 was a new five-membered ring lactone, while compounds 2-5 were isolated for the first time from Eucommia ulmoides bark. Furthermore, compounds 2-4 demonstrated antioxidant activity, with IC₅₀ values for 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging of 46.34 ± 5.80, 74.17 ± 8.71, and 78.02 ± 8.94 μg·mL^-1, and for 2, 2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging of 50.56 ± 1.35, 54.37 ± 3.04, and 55.61 ± 3.41 μg·mL^-1, respectively.

Through repeated application of various chromatographic techniques including silica gel, ODS, MCI, and Sephadex LH-20, 15 compounds were isolated from Magnolia officinalis var. biloba in Lishui. The structures and relative configurations of these compounds were determined using spectroscopic methods such as high-resolution electrospray ionization mass spectroscopy (HRESIMS) and 1D & 2D-NMR. The absolute configurations of compounds (+)-1, (-)-1, (+)-5, and (-)-5 were ultimately established through chiral resolution techniques and quantum chemical calculations of electronic circular dichroism (ECD) spectra. Among these, compounds (+)-1, (-)-1, 2-4, (+)-5, (-)-5, and 6-9 were previously unreported neolignans primarily derived from honokiol or magnolol, while compounds 10-13 were known compounds. The anti-inflammatory activities of all compounds were evaluated using an LPS-induced RAW264.7 cell model. Among them, compound 4 exhibited significant inhibitory activity against nitric oxide (NO) production, with an IC₅₀ value of 1.23 ± 0.04 μmol·L^-1.

In recent years, the synthetic cannabinoid N-(1-amino-3, 3-dimethyl-1-oxobutan-2-yl)-1-butyl-1H-indazole-3-carboxamide (ADB-BUTINACA) has attracted widespread attention due to its high affinity for the CB1 receptor and associated abuse risk; however, its toxicity mechanism remains poorly understood. This study aimed to systematically investigate the toxicological mechanisms of ADB-BUTINACA using an integrated approach. We established an acute rat exposure model (low and high doses: 0.1 and 1 mg·kg^-1) and employed untargeted metabolomics combined with molecular docking analysis to examine its effects on the urinary metabolome and toxicity pathways. Our analysis identified 52 significantly altered metabolites, including taurine and phenylacetaldehyde, and revealed six disturbed metabolic pathways, such as phenylalanine metabolism and one-carbon metabolism. Notably, choline and methionine were pinpointed as potential dose-dependent biomarkers. Molecular docking results demonstrated a strong binding affinity (-7.8 kcal·mol^-1) between ADB-BUTINACA and aldehyde dehydrogenase ALDH3, suggesting that the compound inhibits ALDH3 activity. This inhibition likely disrupts phenylalanine metabolism, leading to the accumulation of phenylacetaldehyde and the upregulation of phenylacetylglycine, thereby inducing oxidative stress and potential cardiovascular and neurotoxicity. Concurrently, the downregulation of methionine and choline indicated an impairment of one-carbon metabolism, which may further exacerbate oxidative stress. In conclusion, this study elucidates the mechanism by which ADB-BUTINACA exposure induces oxidative stress primarily through disrupting phenylalanine and one-carbon metabolism, utilizing non-targeted metabolomics and molecular docking. The identified biomarkers provide a crucial theoretical foundation for future research on the toxicity mechanisms, clinical detection, and risk assessment of synthetic cannabinoids. The experimental protocol complied with the welfare and ethical requirements for experimental animals and was approved by the Animal Ethics Committee of Zhejiang Chinese Medical University (Approval number: 20220718-11).

Yimaikang Soft Capsule is a traditional Chinese patent medicine used to promote blood circulation, remove blood stasis and improve microcirculation in the treatment of ischemic cardio-cerebrovascular diseases. However, its chemical composition and mechanism of action remain unclear. In this study, the chemical constituents of Yimaikang Soft Capsules were systematically analyzed using high-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (HPLC-Q-Exactive Orbitrap MS) and global natural products social molecular networking (GNPS). A total of 125 in vitro components were identified, including 12 compounds reported for the first time in the Yimaikang series of preparations and its raw material herb Erigeron breviscapus. Subsequently, network pharmacology was employed to predict and screen 160 common targets shared between the 125 identified components and the diseases (ischemic cerebrovascular disease and coronary heart disease). Based on these, a "ingredient-target" interaction network and a protein-protein interaction (PPI) network were constructed to identify the core components and key targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed that the mechanism of Yimaikang Soft Capsules primarily involves signaling pathways such as the vascular endothelial growth factor (VEGF) signaling pathway, epidermal growth factor receptor (EGFR) signaling pathway and the advanced glycation end-product receptor (AGE-RAGE) signaling pathway. These pathways are closely associated with biological processes including angiogenesis, inflammation regulation, and apoptosis. Molecular docking validation demonstrated strong binding activity between core components (such as 3, 5, 6, 4′-tetrahydroxy-7-methoxyflavone, aesculin and kaempferol) and their corresponding targets (phosphatidylinositol-4, 5-bisphosphate 3-kinase catalytic subunit beta, PIK3CB; Harvey rat sarcoma viral oncogene homolog, HRAS; phosphoinositide-3-kinase regulatory subunit 1, PIK3R1). This study preliminarily identified the potential active ingredient group of Yimaikang Soft Capsules and elucidated its multi-target, multi-pathway mechanism of action, providing a scientific basis for further clarifying the modern pharmacological foundation of this formulation.

Rapid and accurate identification of chemical components in classical famous formulas is a crucial aspect of their modern research. This study aimed to establish a method for the rapid identification of chemical components in the mass spectrometry data of Xie Bai San by integrating multi-point mass defect filtering (MDF) and molecular networking (MN) techniques, to achieve accurate and comprehensive characterization of its constituents. The mass spectrometric data of Xie Bai San decoction was acquired using ultra-high performance liquid chromatography coupled with quadrupole-Orbitrap tandem mass spectrometry (UHPLC-Q-Orbitrap-MS/MS) in both positive and negative ion modes. The MDF method was employed, utilizing set multi-point mass defect filtering windows to identify structurally similar compounds. The MN approach was applied to create a GNPS molecular network based on the similarity of MS/MS fragmentation patterns, and Cytoscape 3.6.1 software was used to identify clusters of structurally similar molecules, enabling the identification of homologous components via probe molecules. Using the aforementioned methods, combined with retention time, accurate molecular mass, and secondary MS/MS fragmentation characteristics, and based on database and relevant literature information, a total of 185 compounds were identified in the Xie Bai San decoction. These included 54 flavonoids, 42 alkaloids, 17 saponins, 16 stilbenes, 34 organic acids, 15 phenylpropanoids, and 7 other types of compounds. Among these, 46 compounds were reported for the first time in the Xie Bai San formula or its constituent herbs. This study successfully achieved the rapid, accurate, and comprehensive characterization of the chemical components in Xie Bai San decoction. It provides a chemical component foundation for research on the pharmacodynamic material basis and quality control of Xie Bai San compound preparations, and also offers a methodological reference for the rapid identification of chemical components in complex traditional Chinese medicine systems.

This study developed a sustained-release lipid liquid crystalline (LLC) system for leuprolide acetate (LA) and investigated the feasibility of using the lipase inhibitor orlistat (Orl) to modulate the matrix degradation and prolong the drug release period. The LA-LLC formulation was characterized by using polarized light microscopy (PLM), rheological analysis, and in vitro release studies to guide optimization. The impact of Orl on the sustained-release performance of LA-LLC was evaluated through in vitro degradation, in vitro release, and pharmacokinetic studies in rats. All animal protocols were approved by the Laboratory Animals Care and Use Committee of Yantai University (registration number is YTU20210319) (Yantai, China). Results showed that the LLC system utilizing glycerol dioleate (GDO) as the oil phase exhibited optimal sustained-release characteristics. The optimized LA-LLC formulation comprised soybean phosphatidylcholine, GDO, ethanol, and LA at a mass ratio of 44.5∶44.5∶10∶1. This gel exhibited a hexagonal phase structure by PLM. It demonstrated shear-thinning capacity characteristic of a pseudoplastic fluid. In vitro degradation and release results indicated that Orl significantly inhibited lipid degradation and prolonged drug release. Pharmacokinetic results in rats indicated that the addition of inhibitor Orl prolonged the release period from 42 days to 56 days compared to the LA-LLC group. This study demonstrates that the LA-LLC system, with its structural stability and controllable release, can effectively prolong drug release by employing a lipase inhibitor to regulate matrix degradation. The results of the study provide a novel strategy for the sustained delivery of peptide drugs.

Helicide and rosmarinic acid possess sleep-improving properties but suffer from poor aqueous solubility. This study aimed to construct a dual-drug co-loaded amorphous system. Compatibility and intermolecular interactions were predicted using Hansen solubility parameters and molecular docking. Co-evaporates were prepared via the solvent co-evaporation method. Polarized light microscopy, X-ray diffraction, and differential scanning calorimetry analyses confirmed the formation of a single-phase binary co-amorphous system. Fourier-transform infrared spectroscopy, ultraviolet spectroscopy, and nuclear magnetic resonance analyses revealed that hydrogen bonds formed between the aromatic ketone carbonyl group of helicide and the carboxylic acid group of rosmarinic acid, while π-π stacking interactions occurred between their aromatic rings, constituting a stable intermolecular network. Co-amorphization significantly enhanced the apparent solubility of both drugs in pure water by 4.7-fold and 5.1-fold, respectively. In vitro dissolution studies indicated an initial rapid release phase, followed by limitations due to gelation. All animal experiments were conducted in strict accordance with the regulations of the Animal Experiment Ethics Committee of Yunnan University of Chinese Medicine, with approval number: YNUTCM-XMSS-G-20250059. Pharmacodynamic evaluation in a zebrafish insomnia model demonstrated that the co-amorphous system at low and medium doses prolonged total sleep time, increased the number of sleep bouts and average bout length, and shortened sleep latency compared to individual drugs or physical mixture. A combination index (CI) < 0.7 confirmed a synergistic sleep-promoting effect. The solvent co-evaporation method proved efficient and straightforward for preparing the co-amorphous system. This co-amorphous strategy demonstrates significant advantages in improving aqueous solubility and achieving synergistic hypnotic efficacy.

Coreopsis tinctoria Nutt. is an important ethnomedicine traditionally used in Xinjiang, China. This study employed ITS2 barcode sequencing for species authentication of commercial C. tinctoria samples. Furthermore, a comprehensive quality evaluation system was established based on HPLC fingerprinting, quantitative analysis of multiple markers, and chemical pattern recognition. A total of 35 batches of C. tinctoria samples were collected from different growing regions in Xinjiang. Genomic DNA was extracted from leaves, and the ITS2 region was amplified by PCR followed by sequencing. The obtained sequences were assembled, proofread, and validated through BLAST analysis against the NCBI database. Subsequently, a neighbor-joining (NJ) phylogenetic tree was constructed based on the aligned sequences. Chromatographic fingerprinting was performed on a ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm). Similarity analysis of the full-spectrum fingerprints was conducted using the "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (2012 version)". Chemometric approaches including cluster analysis (CA), principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA) were employed. Furthermore, a validated HPLC method was developed for simultaneous quantification of four characteristic constituents. The results showed that the main haplotypes of ITS2 sequences in 35 C. tinctoria samples were H1/H2, with a length of 225 bp. The intraspecific variation was small, and the sequences were relatively conservative, indicating that ITS2 sequences can be used as a reliable DNA barcode for the identification of C. tinctoria. The HPLC fingerprinting showed 16 common peaks, and chlorogenic acid, flavanomarein, marein, and isochlorogenic acid A were successfully identified. The similarity of the samples ranged from 0.936 to 0.998. CA classified the samples into three groups, PCA obtained five principal components with a cumulative variance contribution rate of 84.953%, and PLS-DA identified eight components, including chlorogenic acid, flavanomarein, marein, as markers for regional differences. Quantitative analysis showed that the mass fractions of the four components were as follows: Chlorogenic acid 0.860-7.504 mg·g^-1, flavanomarein 3.368-19.182 mg·g^-1, marein 19.719-83.492 mg·g^-1, and isochlorogenic acid A 2.039-12.568 mg·g^-1. The DNA barcoding technique and integrated HPLC fingerprinting/multi-component method demonstrated high reliability and stability. This study provides a scientific foundation for quality control and further research on C. tinctoria.