Biomedical Chromatography最新文献

Panax notoginseng (P. notoginseng) is one of the most famous natural medicines and widely used to promote blood circulation in health care. However, the active component group of P. notoginseng for activating blood is not clear. We aim to screen and validate the pharmacodynamic component group (PCG), which could exert the same blood-activating effect as P. notoginseng. To clarify the active components, the chemical components were determined by liquid chromatography-tandem mass spectrometry, and the fingerprint of P. notoginseng was established. Twenty candidate active monomers were selected through the spectrum–effect relationship analysis. Eleven active monomers, including Ginsenoside Rg1, Rb1, Rd, F1, Rh1, Rg2, Rb2, Rg3, and Rk1 and Notoginsenoside R1 and R2, were screened out as the PCG through validation by platelet aggregation test. Among them, the antiplatelet aggregation activity of Ginsenoside Rh1 was directly confirmed for the first time. The active component group could exert similar efficacy to the P. notoginseng extract in vitro and in vivo through the validation of in vitro platelet aggregation test and the rats with cerebral ischemia. This study laid the foundation for the quality evaluation of P. notoginseng and provided a reference for the research on the material basis of the pharmacodynamics of other Chinese herbs.

Gualou-Xiebai-Banxia (GXB) decoction shows potential for treating myocardial ischemia (MI), although its underlying mechanism is not fully understood. In this study, a multimodal metabolomics approach, combining gas chromatography–mass spectrometry (GC–MS) and ¹H-NMR, was employed to investigate the cardioprotective effects of GXB in a rat model of myocardial ischemia induced by ligation. ELISA assays and HE staining demonstrated that GXB effectively reduced myocardial injury, oxidative stress markers, and myocardial fibrosis. Orthogonal partial least-squares discriminant analysis identified 62 biomarkers, 20 of which were confirmed using standard compounds. The GC–MS method showed excellent linearity across a wide concentration range (0.004–29.7 μg/mL, R² > 0.9995), with intra- and inter-day precision RSD values below 4.72% and 4.96%, respectively. Method recoveries ranged from 95.40% to 104.83%, with RSD values under 4.84%. Pathway enrichment analysis revealed that GXB decoction alleviates myocardial ischemia-induced damage primarily by modulating energy metabolism and branched-chain amino acid metabolism. These findings provide valuable support for the clinical application of GXB decoction in treating myocardial ischemia.

Previous studies have suggested that ginsenoside Rg₂ glycine ester derivative (RG) exhibits therapeutic potential in mitigating hypoxia. This study aimed to elucidate the potential mechanism of RG in hypoxia injury through a combined approach of metabolomics and network pharmacology. Initially, a CoCl₂-induced cell hypoxia model was established, and the therapeutic impact of RG on biochemical indices was evaluated. Subsequently, metabolomics analysis of cell samples was conducted to identify biomarkers, and network pharmacology was employed to identify potential targets of RG for hypoxia treatment. Finally, the key target and pathway were verified. The study revealed that RG could reverse CoCl₂-induced abnormalities in biochemical indicators. Metabolomics analysis identified 13 biomarkers and seven metabolic pathways associated with RG treatment. Utilizing network pharmacology, five key targets and five metabolic pathways were identified, partially aligning with the metabolomics results. Molecular docking results demonstrated the effective binding of RG to the key targets. Enzyme linked immunosorbent assay verified that RG could exert antihypoxia effect by activated PI3K/Akt pathway. In conclusion, this integrated strategy, combining metabolomics with network pharmacology, sheds light on the protective mechanism of RG against hypoxia-induced cellular damage. The findings offer valuable insights for future research and potential applications of RG in the field.

Praziquantel (PZQ) is the most effective treatment for schistosomiasis, commonly administered as a racemic mixture of the two enantiomers. Despite many reports on the pharmacokinetics of PZQ, the stereoselective pharmacokinetics of PZQ and its major metabolite 4-hydroxypraziquantel (4-OH-PZQ) remain poorly understood in goats. In this study, the chiral LC-MS/MS method was further optimized for separating and quantifying PZQ, trans-4-OH-PZQ, and cis-4-OH-PZQ and their enantiomers and then applied for the molecular pharmacokinetics of three analytes in black goat plasma. The findings showed that PZQ was rapidly absorbed and metabolized to 4-OH-PZQ. The C_max of trans-4-OH-PZQ was about 3 times and 6 times higher than those of cis-4-OH-PZQ and PZQ, respectively. The stereoselectivity of the PZQ and cis-4-OH-PZQ enantiomers was insignificant in black goat plasma (p > 0.05), whereas the trans-4-OH-PZQ enantiomers exhibited obvious stereoselectivity (p < 0.05). The C_max of S-trans-4-OH-PZQ were ~3.1 times higher than that of R-trans-4-OH-PZQ. Further computer simulations indicated that these differences in the stereoselectivity might mainly stem from the different binding energies of the corresponding R- and S-enantiomers of the target analytes to black goat plasma albumin. It has guiding significance for the research on the stereoselectivity of chiral veterinary drugs and their precision medication.

Choerospondias axillaris is a medicinal plant used for treating coronary heart disease (CHD) due to its broad spectrum of anti-inflammatory activities. Cyclooxygenase 2 (COX-2) and lipoxygenase 5 (5-LOX) were immobilized on magnetic nanoparticles for selective ligand-extraction of these two enzymes present in C. axillaris. Sixteen extracted components were identified and analyzed using ultraperformance liquid chromatography plus Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-ExactiveOrbitrap-MS/MS), including flavonoids Curcumin and Epicatechin. The metabolization of the aforementioned 16 components in rats with acute blood stasis indicated a solid pharmacodynamic foundation. This method offers an effective approach for conducting basic research on the enrichment, analysis, and efficacy evaluation of active components in complex natural products.

Vodobatinib is a Bcr-Abl 1 inhibitor, currently entering into Phase 2 clinical trials as a potential drug to treat glioblastoma patients. In the present work, a validated high-performance liquid chromatography (HPLC) detection method for the quantification of vodobatinib in rat plasma was established. Sample preparation involved liquid–liquid extraction method. The chromatographic separation of vodobatinib and the IS was accomplished on an XBridge C₁₈ column using 10 mM ammonium acetate and acetonitrile in gradient condition. The flow-rate and injection volume were 0.8 mL/min and 20 μL, respectively. The eluent was detected at 310 nm. Vodobatinib and the IS eluted at 7.5 and 5.9 min, respectively, with a total run time of 10 min. Validation tests were performed according to FDA requirements. The established method showed good linearity in the range of 50.5–6534 ng/mL. Interday and intraday coefficients of variations were < 5.11%. Stability studies, dilution integrity, and incurred sample reanalysis met the acceptance criteria. Subsequently, the validated HPLC method was used to study the disposition kinetics of vodobatinib in rats.

The aim of this study was to investigate the potential mechanism of Lu-Jiao Fang (LJF) inhibiting endothelial-to-mesenchymal transition (EndMT) in pressure overload-induced cardiac fibrosis. Pharmacokinetic behaviors of the ingredients of LJF were evaluated by LC-MS/MS analysis. Then putative pathways by which LJF regulates EndMT were analyzed by network pharmacology and verified in transverse aortic constriction–induced cardiac fibrosis rats. Loganin, morroniside, salidroside, isopsoralen, and psoralen showed higher plasma, left and right ventricular C_max and AUC_0–t values than hesperidin, specnuezhenide, and icariside II. Twenty-four potential targets related to EndMT were identified, which were mainly involved in relaxin signaling pathway. AKT1, TP53, MMP9, HIF1A, Snail1, and MMP2 were key therapeutic targets in protein–protein interaction network. LJF reversed cardiac dysfunction, left ventricular dilation, and fibrosis and significantly downregulated collagen type I and III and EndMT regulators (Snail1 and Twist1) mRNA expression. In relaxin signaling pathway, the RXFP1 protein expression increased by 22.52%, and the protein phosphorylation of Smad2 and Smad3 decreased by 33.52% and 12.79%, in response to the treatment with LJF. This study initially revealed the EndMT inhibition effects and molecular mechanisms of LJF in cardiac fibrosis, providing a reference basis for the promotion of LJF in the clinic.

Deoxycholic acid (DCA) injection is applied in treating moderate to severe submental bulge or facial fullness caused by excessive submental fat accumulation. Using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) technology, which was swiftly, precisely, and reliably confirmed, DCA was determined in human plasma with low quantification limits of 56 ng/mL. We selected six healthy individual blank human plasma with low concentrations of endogenous DCA and mixed them to prepare standard curve samples. The samples were purified by the protein precipitation technique and then separated using a BEH C18 column (2.1 × 50 mm, 1.7 μm). Using multiple reaction monitoring (MRM) and electrospray ionization (ESI) sources operating in negative mode, the mass was identified and measured. The precursor-to-product ion transitions were observed at m/z 391.2⁻ → 345.2⁻ and m/z 395.2⁻ → 349.2⁻ for DCA and DCA-d₄ (isotope internal standard), respectively. This method was thoroughly validated, encompassing assessments of linearity, sensitivity, precision, selectivity, stability, matrix effect, accuracy, carryover, and recovery. In a word, the validation results demonstrated that this method exhibited sensitivity, accuracy, and reproducibility and could effectively be utilized for studying the pharmacokinetic properties of DCA in a randomized, parallel, controlled, Phase I clinical trial.