Biomedical Chromatography最新文献

A sensitive and robust HPLC-MS/MS method was developed and validated for the simultaneous determination of YWS22026 and its major metabolite M3 in rat plasma using protein precipitation. Chromatographic separation was achieved on a Shim-pack Scepter C18 column (4.6 × 50 mm, 3 μm) with a gradient elution program (mobile phase A: 0.2% formic acid in water; mobile phase B: methanol) at a flow rate of 0.9 mL/min. Detection was performed via positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. The method demonstrated excellent specificity, accuracy (relative error, %RE < 9.1%), precision (relative standard deviation, %RSD < 9.9%), and linearity (5–2000 ng/mL for both analytes). The lower limit of quantification (LLOQ) was 5 ng/mL. Stability studies confirmed that plasma samples remained stable under various storage conditions (at room temperature for 10 h, −20°C for 22 days, three freeze–thaw cycles, and 4°C autosampler storage for 73 h). This validated method was successfully applied to a 26-week concomitant toxicokinetic study in Sprague–Dawley rats, which revealed significant sex-related differences in exposure for both YWS22026 and M3. These findings provide valuable insights for informing the safety assessment of YWS22026 in subsequent clinical trials.

Lomatogonium rotatum (L.) Fries ex Nym. is one of the traditional Mongolian medicines in the “Digeida” category. It has the effects of calming Xieri (a concept in Mongolian medicine) and clearing heat and promoting diuresis and is used for the treatment of diseases such as fever, headache, plague, typhoid, jaundice, and liver and gallbladder heat. However, the mechanism of its flavonoid compounds in the treatment of non-alcoholic fatty liver disease (NAFLD) is not yet clear. In this study, a NAFLD mouse model was established by feeding mice with a choline-deficient high-fat (CDHF) diet for 8 weeks. The relevant indicators in the serum and tissues of mice in each group were measured. The compounds in the L-30 elution fraction were identified by LC-MS technology, and the liver metabolomics was determined. The results showed that the L-30 elution fraction was an enrichment site of flavonoid compounds, and the L-30 elution fraction could effectively alleviate lipid deposition in the liver of NAFLD mice and relieve liver metabolic disorders.

Pharmaceutical excipients have traditionally been regarded as pharmacologically inert carriers, serving merely as vehicles for active pharmaceutical ingredients. However, this conventional paradigm is increasingly challenged by emerging evidence revealing clinically significant drug–excipient interactions that can profoundly influence therapeutic outcomes. This study investigates a previously unrecognized pharmacokinetic interaction between nimodipine, a dihydropyridine calcium channel blocker, and polysorbate 80, a commonly used solubilizing excipient in nimodipine formulations. A novel HPLC–MS/MS method for simultaneous quantification of nimodipine and polysorbate 80 was developed and validated, achieving excellent analytical performance with linearity (r > 0.995), precision (RSD < 15%), and accuracy (RE < 15%). Pharmacokinetic studies in rats revealed that polysorbate 80 co-administration dramatically enhanced nimodipine systemic exposure, increasing AUC by 51.1%, decreasing clearance by 36.5%, and prolonging half-life by 16.1%. Mechanistic investigations using rat liver microsomes demonstrated that polysorbate 80 competitively inhibited nimodipine metabolism with an IC₅₀ of 103.30 μmol/L and K_i of 94.35 μmol/L, while remaining minimally metabolized itself. These findings provide the first evidence of a clinically significant pharmacokinetic interaction between nimodipine and polysorbate 80, challenging the assumption of excipient inertness and highlighting the critical importance of evaluating excipient–drug interactions in pharmaceutical development and clinical practice.

Pravastatin (PRV), a highly hydrophilic statin used to treat hypercholesterolemia and prevent cardiovascular disease, presents certain analytical challenges due to its low bioavailability, tendency to lactonize, and potentially complex metabolism. This review aims to consolidate and critically evaluate the analytical procedures developed for the determination of PRV and its metabolites in pharmaceutical and biological matrices. This review aims to comprehensively summarize four decades of analytical methodologies developed for PRV and its metabolites, comparing chromatographic, spectroscopic, electrochemical, and capillary electrophoresis approaches in terms of sensitivity, selectivity, and matrix applicability. This review also evaluates essential bioanalytical nuances, including PRV's lactone–acid interconversion, matrix-induced ion suppression, pH-dependent instability, low ng–pg·mL⁻¹ plasma levels, and metabolite-specific detection challenges alongside pharmaceutical analytical considerations. The literature demonstrates that liquid chromatography–tandem mass spectrometry and high-performance liquid chromatography remain the most sensitive and reliable platforms for ultratrace bioanalysis, while spectrophotometric and electrochemical methods provide cost-effective alternatives for formulations and higher concentration samples, and capillary electrophoresis offers efficient separation with low solvent use. Future analytical advances should prioritize metabolite-resolved quantification, green extraction strategies, microfluidic and point-of-care designs, and AI-assisted data processing to improve sensitivity, stability assessment, and high-throughput monitoring of PRV in clinical and environmental settings.

Microsorum fortunei (T. Moore) Ching (MF) is a traditional Chinese medicine utilized for the treatment of rheumatoid arthritis (RA). However, the chemical composition and underlying mechanisms of MF remain unclear. This study aims to identify the active ingredients of MF that contribute to its therapeutic effects on RA and to explore the associated mechanisms. A rat model of RA was established, and the effects of MF were evaluated by monitoring changes in appearance, body weight, inflammatory indices, joint histopathology, thymus and spleen indices, and levels of inflammatory factors. The ethyl acetate fraction of MF (EAF-MF) was analyzed using UPLC-Q-TOF/MS. Network pharmacology and molecular docking analyses were conducted to predict the anti-inflammatory components and pathways. The EAF-MF significantly alleviated RA symptoms, leading to reductions in foot swelling, arthritis index, joint damage and pro-inflammatory cytokines. UPLC-Q-TOF/MS identified 57 compounds within the EAF-MF; network pharmacology identified 44 active components connecting 524 targets. The data showed that MF treatment of RA was related to oxytocin signaling pathway and Cushing syndrome. Molecular docking confirmed strong binding between core targets and key components. Consequently, this study elucidates the multi-component, multi-target, and multi-pathway mechanisms of EAF-MF in the treatment of RA, thereby supporting its clinical application.

Intervertebral disc degeneration (IVDD) is a major cause of low back pain and disability. Excessive reactive oxygen species (ROS) disrupt disc homeostasis. Bushen Huoxue (BSHX) decoction, a traditional Chinese medicine, shows clinical efficacy in IVDD, but its antioxidant mechanisms are not well understood. Public IVDD transcriptomic datasets were analyzed to identify ROS-related gene signatures, followed by immune infiltration assessment. LC–MS/MS was used to characterize the chemical profile of BSHX decoction. Network pharmacology, machine learning, and bioinformatics analyses were integrated to identify potential compound-target interactions. Molecular docking and molecular dynamics simulations were applied to validate binding stability between active components and key targets. Six ROS-related hub genes (NCF1C, SLC11A1, MPO, GSTA4, SLC7A11, AKR1C3) were identified, with immune profiling revealing increased neutrophils, NK cells, and pro-inflammatory macrophages in IVDD. LC–MS/MS identified 77 active compounds in BSHX decoction, including flavonoids and phenolic acids. Network analysis and docking suggested strong interactions with oxidative stress targets, notably MPO. Molecular dynamics simulations confirmed the stability of key compound-target complexes. BSHX decoction shows promise as a multitarget antioxidant therapy for IVDD, offering mechanistic insight into its traditional efficacy. These findings support its further development and experimental validation.

The aim of this study is to further clarify the material basis and mechanism of the QianLieXin (QLX) capsule. UPLC-Q-MS was employed to identify the active substances absorbed into the blood and prostate. Pharmacological network analysis and anti-inflammatory assays were performed. Twenty principal bioactive compounds of the QLX capsule were identified in the blood and prostate tissues of rats. Network pharmacology and molecular docking analyses demonstrated 292 potential targets of these active substances relevant to the treatment of chronic prostatitis. Among them, chlorogenic acid, apigenin, kaempferol, isoquercitrin, and ursolic acid were identified as the primary agents exerting anti-inflammatory effects. The principal molecular targets included AKT1, EGFR, PIK3, and MAPK, among others. Key signaling pathways encompassed the neuroligin-receptor interaction pathway as well as lipid metabolism and atherosclerosis-related pathways. Experimental results demonstrated that both the medicated serum and the active substances significantly suppressed lipopolysaccharide-induced interleukin-1β levels, inhibited the expression of NF-κB protein, and reduced reactive oxygen species production, indicating notable anti-inflammatory and antioxidant activities. Furthermore, QLX has a strong binding effect on targets such as EGFR, AKT, and MMP9. Medicated serum was shown to downregulate the EGFR/AKT/MAPK/MMP9 signaling pathways. Our findings clarified the pharmacodynamic substances of the multicomponent QLX capsule and its regulatory mechanism.