Biomedical Chromatography最新文献

Serum pharmacochemistry was applied to identify the blood-absorbed constituents of dry ginger (DG) and processed ginger (PG). Network pharmacology was employed to construct compound–target–pathway interaction networks, which were further validated through molecular docking to assess the binding affinities between ligands and key targets. Furthermore, a dextran sulfate sodium (DSS)-induced murine model of ulcerative colitis (UC) was established to systematically investigate the multi-target regulatory mechanisms of DG and PG. The processing of DG transforms gingerols into shogaols, thereby modifying its bioactive composition and physiological functions. DG, which is rich in 6-gingerol, mitigates intestinal inflammation through the inhibition of the PI3K-Akt signaling pathway, whereas PG increases the levels of shogaols and zingerone, promoting gut barrier restoration and reducing occult bleeding. Our findings illustrate how processing alters the functional properties of ginger, offering valuable insights for the development of ginger-derived products tailored to specific gastrointestinal health benefits.

Zhi-Zi-Hou-Po Decoction (ZZHPD) is a classic traditional Chinese medicine (TCM) formula for treating depression and anxiety. Considering the disadvantages of decoction, our research group prepared ZZHPD into Zhi-Zi-Hou-Po Granule (ZZHPG). However, the chemical composition and mechanism of ZZHPG for the treatment of depression–anxiety comorbidity remain unclear. In this study, the UHPLC-Q-TOF-MS technology was applied to investigate the chemical constituents in ZZHPG, and network pharmacology was used to reveal the mechanism of ZZHPG. A total of 92 compounds were identified in ZZHPG. The network pharmacological analysis showed that 92 compounds regulated 132 pathways through 90 targets, among which 17 compounds maybe the material basis of ZZHPG for the treatment of depression–anxiety comorbidity, and osthole, obovatol, and naringenin may be the main pharmacodynamic components. Further research indicated that the mechanism of ZZHPG in treating depression–anxiety comorbidity may be related to the inhibition of nervous system disorders, activation of GABA receptors, anti-inflammation, and reduction of oxidative stress. The molecular docking results showed that osthole, obovatol, and naringenin exhibited strong binding affinity with the core targets AKT1, ALB, TNF, EGFR, ESR1, and GSK3B. This research laid foundation for clinical application of ZZHPG and provided new ideas for the study of other TCM prescriptions.

A stability-indicating HPLC method was developed to quantify belzutifan and its related and degradation impurities, and LC–MS/MS technology was used for the characterization of stress-induced degradation products (DPs). Adequate separation was attained on an waters X Bridge C18 column with dimensions 150 × 4.6 mm (3.5 μm) at 30°C, utilizing acetonitrile and 0.1% formic acid in water (60:40) v/v as the mobile phase. This method was set at a flow rate of 0.8 mL/min, and a PDA detector was used for detection. The method demonstrated excellent specificity, accuracy (98.24%–101.01% recovery), precision (%RSD < 2), linearity (R² > 0.999), and sensitivity, with resolution > 2.0. Forced degradation studies showed no degradation under UV and thermal stress but significant degradation under acidic, basic, and oxidative conditions, yielding two acid-induced (DP A/B-1 and A-2), one base-induced (DP A/B-1), and one peroxide-induced (DP O-1) DPs. The degradation pathways of DPS were established by using the mass fragmentation patterns. This method represents stability, is compliant with ICH guidelines, and is well suited for routine quality control and comprehensive stability assessment of belzutifan. A representation of the stress study of belzutifan is shown.

Monothioglycerol predominantly acts as an antioxidant in injectable solutions, inhibiting the oxidation and degradation of medications that are sensitive to oxygen, alongside other formulation ingredients. It scavenges free radicals and ceases the chemical chain reactions that result in instability. A thorough, swift, straightforward, and eco-friendly stability-indicating liquid chromatography (LC) method has been developed and validated, facilitating the measurement of monothioglycerol concentrations in injectable solutions. The antioxidant was studied through several tests in accordance with ICH guidelines, including characteristics such as forced degradation studies. Through isocratic separation, monothioglycerol and its related degradants were effectively resolved. The antioxidant was determined using an end-capped octadecylsilyl (C18) column, and the method was validated under room temperature conditions. The isocratic mobile phase operates at a flow rate of 1.0 mL/min and consists of fixed ratios of filtered water, acetonitrile, and orthophosphoric acid. The overall AES grade of 86, AGREE grade of 0.64, MoGAPI grade of 76, BAGI grade of 75.0, and ultimate whiteness of 95.8 all highlight the ecological advantages of the HPLC method. The current research indicates that the proposed methodology is unique, precise, accurate, stable, and environmentally beneficial at a microscale, making it suitable for routine quality control analysis.

Metoclopramide (MCP) is a commonly used prokinetic and antiemetic medication prescribed for managing gastrointestinal motility disorders, nausea, and vomiting. The risk of adverse reactions, as well as accurate quantification of MCP in pharmaceutical products and biological samples, is essential for quality control, pharmacokinetic assessment, and therapeutic monitoring. Relevant studies for this review were gathered from major scientific databases, including Scopus, Web of Science, ScienceDirect, Google Scholar, and PubMed, restricted to publications written in English. This review provides a critical overview of analytical techniques reported over the last 25 years for MCP determination, focusing on chromatographic, spectroscopic, and electroanalytical methods. Chromatographic techniques—particularly high-performance liquid chromatography (HPLC)—remain the gold standard due to their robustness, precision, and compatibility with diverse sample matrices. Advanced LC–MS/MS systems offer exceptional sensitivity, reaching pg–ng/mL levels, which is particularly valuable in bioanalytical applications. HPLC-UV approaches, though less sensitive, are cost-effective and suitable for routine quality assessment. Spectroscopic methods such as UV–Vis spectroscopy are rapid and affordable but limited by poor selectivity in complex matrices. Electrochemical and voltammetric methods have recently gained attention, offering low cost, speed, and remarkable sensitivity, especially with nanomaterial-modified electrodes. Remaining challenges include MCP instability, matrix interferences, and balancing sensitivity with accessibility.

Cnidii Fructus (CF) is a Chinese herbal medicine with bioactive immune properties and potential applications in treating ulcerative colitis (UC), but its well-defined bioactive constituents are understudied. The current study aimed to investigate CF's anti-UC bioactivity and elucidate its bioactive constituents via chromatographic analysis, in vitro/in vivo tests, and spectrum-effect analysis. Results showed 10 batches of CF inhibited the inflammatory response induced by lipopolysaccharide (LPS)–stimulated RAW 264.7 macrophages. High-performance liquid chromatography (HPLC) fingerprint profiles of the 10 batches were established, with 16 common peaks identified. Subsequently, 15 of these peaks (including eight reference-validated compounds) were characterized by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS). Four anti-UC-related compounds were identified via spectrum-effect relationships, namely, diosmin, xanthotoxol, imperatorin, and osthole. Experimental verification confirmed these as key components potentially linked to alleviating inflammatory responses in UC. This study is the first to establish a spectrum-effect relationship for CF's anti-UC effects, identify its critical active constituents, and provide a scientific basis for CF quality control and UC therapeutic development.

The leaves of Dimocarpus longan Lour. (LYY), a plant belonging to the Sapindaceae family, are a traditional Chinese medicine (TCM) with Guangxi national characteristics. However, its quality standards remain inadequate, and considering the impact of the current standard harvest period on medicinal plants, it is necessary to study how different harvest periods affect the component contents of LYY. In this study, high-performance liquid chromatography (HPLC) combined with chemometrics was employed to analyze the chemical components of LYY across various harvest periods. By comparing 12 batches of LYY fingerprints, 8 compounds were identified. The results of similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and weighted technique for order preference by similarity to ideal solution (TOPSIS) revealed differences in LYY samples from different harvest periods, with the samples of January and February being of better quality, finding that gallic acid, quercitrin, and quercetin might be potential markers for these compositional variations. Quantitative analysis demonstrated that the contents of different compounds varied with the harvest period. In conclusion, the established method can be applied to comprehensively evaluate the LYY quality, providing a scientific basis for its reasonable harvest.

The growing resistance of Plasmodium species to existing antimalarial drugs and the limited global supply of artemisinin pose significant challenges to malaria control programs. Since Artemisia annua is currently the sole commercial source of artemisinin, there is a pressing need to identify supplementary sources. The present study investigates Artemisia tournefortiana, a plant native to the Trans-Himalayan region of Ladakh, as an alternative source of artemisinin. Quantification of artemisinin in different plant parts was carried out using RP-HPLC-PDA with a C18 column (250 mm × 4.6 mm, 5 μm) and a mobile phase comprising phosphate buffer (pH 2.2) and acetonitrile (50:50, v/v). The maximum artemisinin content was detected in the flowers (1.43%), followed by roots (0.33%) and seeds (0.32%), with flower content varying from 0.05% to 1.43% across different geographical sites. Gene sequences encoding enzymes involved in the artemisinin biosynthetic pathway were retrieved from GenBank, and specific primers were designed using Primer3 for comparative DNA sequencing of A. annua and A. tournefortiana. Sequence comparison revealed notable similarities, supporting the biosynthetic potential of A. tournefortiana as an alternative, though less productive, source of artemisinin.

This study aims to analyze secondary metabolites of Dendrobium fimbriatum via UPLC-MS/MS; establish hyperthyroidism-induced kidney yin deficiency model with thyroid tablets to study its decoction's effects; use UPLC-MS/MS untargeted metabolomics to screen blood-entry components, biomarkers, and potential active substances; explore component–target–pathway mechanism via network pharmacology and molecular docking. One hundred thirty-eight secondary metabolites are identified. Decoction increased rat body weight/pain threshold, reduced temperature; alleviated kidney yin deficiency symptoms/pathology; regulated thyroid (T3/T4), gonad (T/E2), and nucleotide (cAMP/cGMP) functions; and changed OPLS-DA profile; 13 blood-entry components, 30 biomarkers, and 9 potential “nourishing kidney yin” substances are found—four high-binding components act on EGFR/AKT1/SRC via cancer pathogenesis, endocrine resistance, and relaxin pathways. Decoction improves model rats' metabolism; four active components (e.g., N-p-coumaroyltyramine) regulate EGFR/AKT1/SRC via key pathways, affecting biomarkers and glycerophospholipid/amino acid/linoleic acid metabolism to exert anti-inflammatory/antioxidant/antitumor/antiapoptotic effects, realizing “nourishing kidney yin”; this study supports mechanism analysis, quality evaluation, and new drug R&D of D. fimbriatum.

Ephedra and Houttuynia Herb Powder (MHYXD) is a traditional Chinese herbal formula widely used in veterinary medicine to treat respiratory infections, such as mycoplasma pneumonia in calves, although its active components and pharmacological mechanisms have remained unclear. To address this, the present study identified the bioactive compounds in MHYXD using LC–MS, which revealed a total of 32 active compounds, with ephedrine, quercetin, and houttuynin being the most prominent. Network pharmacology and molecular docking approaches were employed to predict potential targets and pathways, indicating that MHYXD acts on multiple targets—including TNF-α, IL-6, and TLR4—and regulates the NF-κB and MAPK signaling pathways; molecular docking further confirmed strong binding affinities between key compounds and these target proteins. The antimycoplasma and anti-inflammatory effects of key components were validated through in vitro and in vivo experiments, which demonstrated that MHYXD significantly reduced mycoplasma load and alleviated lung inflammation in calves. In conclusion, this study elucidates the multicomponent, multitarget, and multipathway mechanisms of MHYXD in treating Mycoplasma pneumonia, thereby providing a scientific basis for its clinical application in veterinary medicine.