Biomedical Chromatography最新文献

Polygonum orientale L. (PO) represents significant bioactivities in treating myocardial ischemia (MI); however, its underlying mechanisms remain unclear. This study aims to elucidate PO's potential mechanisms in MI using an integrated approach that combines UHPLC–Q-Exactive Orbitrap HRMS, network pharmacology, and RNA-sequencing (RNA-seq). Initially, the chemical constituents of PO were identified using UHPLC–Q-Exactive Orbitrap HRMS. Subsequently, network pharmacology, molecular docking, and RNA-seq were employed to screen potential active components of PO targeting MI and predict their molecular mechanisms. Then, the molecular mechanisms were verified using western blotting and ELISA in MI mice. A total of 45 components were identified from PO, with 14 potential active compounds interacting with 204 MI-related genes. The findings suggest that PO could alleviate heart damage. The RNA-seq results indicated 244 potential targets regulated by PO. Integrating RNA-seq and network pharmacology analyses revealed that the toll-like receptor signaling pathway plays an important role, alongside the PI3K-Akt. Notably, PO reduced the expression of TLR4 and TLR2 while increasing p-Akt and p-PI3K levels in MI mice, leading to decreased inflammatory cytokines and apoptosis-related proteins. This study provides initial evidence that PO inhibits the toll-like signaling pathway and activates PI3K–Akt signaling pathway to exert protective effects against MI.

Oleracrylimide E, a new alkaloid from Portulaca oleracea L., shows significant bioactivity. Then, the pharmacokinetics of this alkaloid after its intravenous administration at dose of 2.0 mg/kg body weight in rat plasma was evaluated using UHPLC with vitexin-2″-O-rhamnoside as an internal standard (IS). The UHPLC-ESI-Q-TOF/MS method was applied to investigate the metabolism of Oleracrylimide E in rats in response to the immediate conversion of Oleracrylimide E into other metabolites after its oral administration. As a result, a total of 16 metabolites of Oleracrylimide E were identified in the plasma, urine, and fecal of the experimental rats.

Bimiralisib, a pan-PI3K/mTOR inhibitor, has demonstrated antitumor efficacy in preclinical models. In this study, we present a validated LC-MS/MS method for quantifying bimiralisib from dried blood spots (DBS) in mice. The method was validated in accordance with FDA guidelines. Bimiralisib was extracted from DBS disks using a liquid–liquid extraction technique. Chromatographic separation was achieved on an Atlantis dC18 column (100 × 4.6 mm) using an isocratic mobile phase. The flow rate was set at 0.70 mL/min. Under optimized conditions, the retention times for bimiralisib and the internal standard (IS, Nilotinib) were approximately 1.14 and 1.27 min, respectively, with a total run time of 2.00 min per injection. The monitored MS/MS ion transitions were m/z 412.2 → 141.0 for bimiralisib and 530.4 → 259.0 for the IS. The method employed a broad calibration range (1.00–1434 ng/mL) with a determination coefficient (r²) of 0.996. All validation parameters met the required acceptance criteria, and hematocrit levels had no impact on bimiralisib concentrations in DBS. The validated method was utilized to determine intravenous and oral pharmacokinetic parameters by quantifying bimiralisib in mouse blood, with results correlated to pharmacokinetic data from mice plasma.

The development of novel active molecules with a clear target has always been an urgent need for the treatment of epilepsy. Previously, we reported 2-(2-fluorophenyl)-5-phenyl-7-propyl-[1,2,4]triazolo[1,5-a]pyrimidine (10C) as a selective and positive modulator of GABA_A receptors, which exhibited excellent antiepileptic activity in mice with an ED₅₀ value of 8.51 mg/kg. However, the pharmacokinetics (PK) profiles of this compound remain unclear. In this study, 10C and four analogs (10A, 10B, 10D, and 10E) as well as deuterated 10C were synthesized with high efficiency under optimized reaction conditions, and deuterated 10C was employed as an internal standard. The concentrations of the five compounds in rat plasma and of 10C in tissue homogenate were assayed using a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method. The results indicated that, compared to the other four analogs, 10C exhibited the highest drug concentration in rat plasma at the same dosage, which provides a good explanation for its superior antiepileptic activity compared to the other four analogs. Following intraperitoneal injection, 10C displayed favorable pharmacokinetic characteristics (F = 41%) and excellent brain penetration potency (B/P = 1.9). Overall, compound 10C is a promising lead for the research and development of new small-molecule therapeutics for epilepsy.

Xuanbi Decoction (XBD) is a classical traditional Chinese medicine (TCM) effective in treating different types of arthritis. This study aimed to integrate metabolomics with network pharmacology to identify active metabolic components of XBD, elucidate its therapeutic targets, and reveal the key signaling pathways involved in the treatment of gout. The study systematically analyzed the material basis and potential mechanisms underlying XBD efficacy in gouty arthritis (GA). First, 352 blood metabolites from XBD were screened by extracting the drug-containing serum and utilizing liquid chromatography–tandem mass spectrometry (LC-MS/MS). Twenty-two key ones were identified through correlation analysis. Two-hundred fifty-five metabolite-related targets and 764 GA-related targets were retrieved from multiple databases. Further analysis of the intersection of targets identified 60 key overlapping targets. PPI network analysis elucidated the interrelationships among the 60 targets. GO and KEGG pathway enrichment analyses were conducted on these crossover targets, identifying 25 GO terms and 20 KEGG pathways. Network diagrams were constructed, featuring “22 metabolites–60 targets–25 GO terms” and “22 metabolites–60 targets–20 KEGG pathways.” Additionally, a comprehensive network map was constructed, featuring “9 XBD drugs–22 active metabolic components–60 core targets–25 signaling pathways,” elucidating the multidimensional intervention mechanism of XBD on GA, offering insights into its clinical application in GA treatment.

Disorders of consciousness (DOC), including unresponsive wakefulness syndrome (UWS) and minimally conscious state (MCS), are complex brain dysfunctions with various causes. Misdiagnosis is common when relying solely on neurological exams, highlighting the need for accurate differentiation to guide clinical rehabilitation. This study explores metabolomic differences between UWS and MCS across cerebrospinal fluid (CSF), serum, and urine samples to identify biomarkers and metabolic pathways. Fifty-one subjects were categorized into UWS (n = 35) and MCS (n = 16) based on coma recovery scale-revised (CRS-R) scores. Ultraperformance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) was used to analyze samples, and statistical methods identified 14, 24, and 22 differential metabolites in CSF, serum, and urine, respectively. CSF metabolites were linked to necrosis, apoptosis, and neuroprotection; serum metabolites to lipid metabolism and immune response; and urine metabolites to cell signaling and neural function. Metabolomic panels showed AUC values of 0.85 (95% CI: 0.73–0.96) for CSF (95% CI: 0.86–1.00), 0.94 for serum, and 0.93 (95% CI: 0.79–1.00) for urine in distinguishing UWS from MCS. This profiling offers valuable insights into DOC pathophysiology and aids in accurate differentiation of these states.

Taxillus chinensis (DC) Danser (T. chinensis) is broadly used in traditional Chinese medicine. Although pharmacological research shows that the ethyl acetate extract of T. chinensis (JEA) has beneficial effects in treating hyperuricemic nephropathy (HN), the active components and potential mechanisms for these effects remain unclear. This study aims to predict the effective components and mechanism of JEA for HN. Firstly, we adopted UHPLC-Q-Exactive HFXMS technology to analyze the chemical profile of JEA and the absorbed prototype ingredients in rat plasma. In addition, network pharmacology methods were utilized by us to elaborate on the active compounds, signaling pathways, and potential mechanisms of JEA in treating HN. Finally, a UPLC method was established to screen potential chemicals that can effectively inhibit the activity of xanthine oxidase (XOD). A total of 92 components were systematically characterized in JEA. Of those, 46 compounds were identified in the plasma of rats administered with JEA extract. Through network pharmacology, 10 potential active components, 10 crucial target genes, and 20 pathways were predicted to be involved in the JEA-mediated treatment of HN. The molecular docking results indicated that oxyresveratrol, isorhammeiol, and robinetwere exhibited strong binding affinities for GAPDH, PPARG, and ALB. The analysis of the XOD inhibition experiment suggested that dihydromyricetin and oxyresveratrol exhibited potent inhibitory effects on XOD, with IC₅₀ values of 0.48 and 0.68 mg·mL⁻¹. This study preliminarily identified the potential effective components of JEA and revealed its underlying molecular mechanisms of action in preventing HN, which will improve our further studies on JEA to treat HN.

Type 2 diabetes mellitus (T2DM) is characterized by pancreatic β-cell dysfunction and insulin resistance, with pyroptosis emerging as a key contributor to β-cell loss. Jiedu Tongluo Tiaogan Formula (JTTF), a traditional Chinese medicine (TCM), has shown clinical efficacy in T2DM management, but its mechanism linking pyroptosis remains unexplored. This study integrates UPLC-MS/MS, network pharmacology, and in vivo experiments to elucidate JTTF's anti-diabetic mechanisms. UPLC-MS/MS identified 441 compounds in JTTF, predominantly alkaloids, flavonoids, phenols, and terpenoids. Network pharmacology revealed JTTF's multi-target effects on T2DM-associated pyroptosis, particularly via the NLRP3/Caspase-1/GSDMD pathway. In diabetic mice, JTTF dose-dependently reduced fasting blood glucose, insulin resistance, and dyslipidemia, while restoring pancreatic β-cell morphology. Mechanistically, JTTF suppressed NLRP3 inflammasome activation, downregulated Caspase-1 and GSDMD expression, and attenuated IL-1β/IL-18 release. Notably, this study provides the first evidence of JTTF's anti-pyroptotic effects in T2DM, highlighting its unique ability to modulate glycolipid metabolism and inflammatory cell death concurrently. These findings underscore JTTF's translational promise for preserving β-cell function and suggest future exploration of non-classical pyroptosis pathways. Our work bridges traditional medicine and molecular pharmacology, paving the way for clinical trials and integrative T2DM therapies.