Biomedical Chromatography最新文献

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.

Panax ginseng hydrosol, a water-distilled coproduct of ginseng extraction, contains diverse volatile and semi-volatile terpenoids beyond the well-studied ginsenosides. We applied HS-SPME-GC–MS semiquantitative analysis–based network pharmacology approach to systematically characterize the multi-target and multi-pathway potential of 30 principal hydrosol constituents. In silico target prediction (BATMAN-TCM) identified 78 human protein targets, which were integrated with high-confidence protein–protein interactions. Ginsenol is the main volatile component in Panax ginseng hydrosol, followed by sandaracopimarinol, (+)-spathulenol, (−)-spathulenol, isospathulenol, (+)-cycloisolongifol-5-ol, and rosifoliol. Minor components include monoterpenes such as alpha-terpineol and menthol, alcohols, ketones, and hydrocarbons like (−)-globulol and ferruginol. For the top 5 diseases and indications, most strongly supported as target gene explanations based on main components and robust database-backed evidence, pain modulation, neurodegenerative diseases, breast cancer, androgen receptor-related disorders, and autoimmune–metabolic genetic disorders emerge as leading categories. These findings validate a multicomponent, multi-target mechanism underlying Panax ginseng hydrosol's traditional uses and highlight synergistic interactions that extend the pharmacological landscape beyond ginsenoside-centric models. Our study provides a molecular framework for future experimental validation and rational development of hydrosol-based therapeutics.

Endophytes are microorganisms that reside inside the living tissues of leaves and possess antimicrobial, antioxidant, anticancer and anti-inflammatory activity etc. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) represents a powerful analytical platform for comprehensive metabolomic profiling of endophytic fungi, offering superior sensitivity, selectivity and structural elucidation capabilities compared to conventional methods. This study employed LC–MS/MS and GC–MS analyses to characterise bioactive compounds from the methanolic extract of endophytic fungus Acremonium sclerotigenum CBS 124.42 isolated from Stevia rebaudiana leaves. LC–MS/MS analysis identified nine major bioactive compounds, including flavonoid glycosides (928.4 m/z), rutin derivatives (1454.7 m/z), cyclic depsipeptides (1445.4 m/z), cyanidin-3-O-rutinoside (595.3 m/z), macrolactams (1394.2 m/z), proanthocyanins (1232.8 m/z), oligomeric imperatorin (676.8 m/z) and cyanidin-3-O-glucoside (1232.8 m/z). Stevioside was detected at 9.02 min with characteristic fragments at 317.9 m/z. GC–MS analysis revealed 18 distinct compounds, with benzene (azidomethyl) being most abundant (100% relative area), followed by hydrazine carbothioamide (71.3%) and benzyl(1,2,3-thiadiazol-4-yl) carbamate (43.22%). Significantly, this study reports the first identification of hydrazine carbothioamide and benzyl(1,2,3-thiadiazol-4-yl) carbamate from natural sources, compounds previously known only from chemical synthesis. These compounds demonstrated significant bioactivities including antioxidant (IC₅₀ = 28.1 ± 6.96 mg/g), antimicrobial (zone of inhibition: 10.3–23.6 mm) and anticancer properties, establishing A. sclerotigenum CBS 124.42 as a novel source of therapeutically relevant secondary metabolites.

This study used untargeted metabolomics via UHPLC-Q-Orbitrap HRMS to compare chemical composition differences between the Chinese medicinal herb Spatholobus suberectus Dunn (Jixueteng) and its common adulterants (Mucuna macrocarpa, Callerya dielsiana, Kadsura heteroclita), aiming to clarify their metabolic profiles. Multivariate statistical analyses (PCA, OPLS-DA), volcano plot analysis, and KEGG pathway enrichment were employed to identify differential metabolites and related pathways. A total of 1157 compounds were identified, predominantly flavonoids, followed by phenolic acids and alkaloids. Significant differential metabolites were screened using VIP > 1 and p < 0.05. For S. suberectus vs. M. macrocarpa and C. dielsiana, differential metabolites were mainly enriched in flavonoid and anthocyanin biosynthesis pathways. In contrast, those between S. suberectus and K. heteroclita concentrated in four pathways: metabolic pathways, secondary metabolite biosynthesis, flavonoid biosynthesis, and flavone/flavonol biosynthesis. These findings reveal interspecific metabolic profile differences, laying a data foundation for developing characteristic chemical marker-based identification techniques. This supplements existing methods and enhances medicinal material quality control.

To develop an LC–MS/MS method for simultaneous quantification of thioridazine and its metabolites in rat plasma, samples were prepared by protein precipitation and filtration using thioridazine-d3 as an internal standard. Chromatographic separation was achieved on a reversed-phase C18 UPLC column with 0.1% formic acid in acetonitrile and 10-mM ammonium acetate in water at 0.7 mL/min. Quantification was performed using multiple reaction monitoring in positive ion mode with the following precursor–product ion transitions: m/z 371.1 → 126.1 (thioridazine), m/z 387.1 → 126.1 (mesoridazine), m/z 403.1 → 126.1 (thioridazine-2-sulfone), m/z 387.2 → 126.1 (thioridazine-5-sulfoxide), and 374.1 → 129.1 (thioridazine-d3). This method showed good linearity: 0.1–1000 ng/mL for thioridazine (r² = 0.9980) and 0.5–1000 ng/mL for mesoridazine (r² = 0.9981), thioridazine-2-sulfone (r² = 0.9976), and thioridazine-5-sulfoxide (r² = 0.9967). The accuracy of the LLOQ and QC samples for thioridazine and its metabolites was within 85%–115%, with intrabatch and interbatch precisions (CV%) below 15%. The average recovery ranged from 98% to 109%, and no significant matrix effect was observed, with values ranging from 93% to 110%. All analytes were stable under the conditions used in this study. This method was applied in a pharmacokinetic study following the intravenous administration of thioridazine to rats.

A highly sensitive and versatile liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed for the simultaneous quantification of upadacitinib and atorvastatin in rat plasma. The analytes and internal standard were extracted from 30 μL rat plasma using a simple protein precipitation technique. The method demonstrated excellent linearity over the concentration ranges of 0.5–500 ng/mL for upadacitinib and 0.05–100 ng/mL for atorvastatin and precision over all quality control levels. Detection was achieved using multiple reaction monitoring (MRM) in positive ionization mode. Method validation was carried out in accordance with the current ICH M10 guidance on bioanalytical method validation. The validated method was successfully applied to a pharmacokinetic study of a 10 mg oral formulation of upadacitinib and atorvastatin in rats. The pharmacokinetic results confirmed the applicability of the method for the reliable and simultaneous determination of both analytes in rat plasma.

Glutathione (GSH), renowned for its antioxidant properties and ability to inhibit tyrosinase activity, finds widespread application in skincare products for its anti-aging and skin-whitening effects. However, the presence of thiol (-SH) groups in glutathione imparts an unpleasant odor, detracting from user comfort. In this study, we investigate the potential of ginsenosides (GS) derived from Panax ginseng C.A. Meyer to mask the malodor of glutathione in cosmetics. We validate the odor masking effect of GS on GSH through HS-SPME-GCMS technology. Through multispectroscopic analyses including Fourier transform infrared spectroscopy (FTIR) and fluorescence spectroscopy, as well as UPLC/QTOF-MS detection, we elucidate the formation and molecular mechanisms of the GSH-GS complex. Our findings reveal intermolecular interactions between glutathione and GS, leading to alterations in spatial conformation and molecular polarity of GSH. Furthermore, fluorescence assays demonstrate the quenching of glutathione fluorescence by GS via a static mechanism. Thermodynamic evaluations confirm the spontaneity of complex formation and provide insight into the bonding forces involved. The formation of the GSH-GS complex increased the transdermal absorption rate of GSH in vitro. Overall, the GSH-GS complex presents a promising strategy for enhancing the olfactory profile of skincare products containing glutathione, thereby improving consumer acceptance and efficacy.

Malaria, as a serious parasitic infectious disease, remains a major threat to global public security. This study investigated the metabolic mechanisms of patchouli alcohol (PA) and artemisinin sodium (SA) in treating malaria using pharmacodynamics and metabolomics. A Plasmodium berghei ANKA (PbA)–infected mouse model was established to evaluate the antimalarial effects of PA and SA combination therapy (SP). Pharmacodynamic results showed that the SP exhibited better therapeutic effects in infection suppression, weight recovery, and reduction of malaria pigment deposition compared with individual treatments. Untargeted metabolomics identified 146 plasma differential metabolites, including fatty acids, phospholipids, and inflammation-related factors, linked to pathways such as unsaturated fatty acid biosynthesis and glycerophospholipid metabolism. Tetradecanedioic acid (TDA) was identified as a potential biomarker, reflecting the drug interactions and therapeutic efficacy. TDA was quantified by targeted metabolomics to study the interactions of combination drugs. The findings suggest that PA and SA synergistically enhance antimalarial effects by modulating lipid metabolism and anti–inflammatory properties and influencing membrane fluidity. This study provides insights into the metabolic mechanisms of PA and SA combination therapy, supporting its clinical application as a novel antimalarial adjuvant.