Biomedical Chromatography最新文献

An increased risk of developing respiratory diseases has been linked to exposure to cigarette smoking (CS). The flower buds of Tussilago farfara L., also known as Farfarae Flos (FF), can be used for the treatment of cough, bronchitis, and asthmatic disorders in China. In the present study, we used lung and fecal metabolomics, as well as the intestinal flora analysis, aimed to investigate the protective effect of FF against the CS exposure induced lung inflammation on mice. The results showed that FF administration could relieve the lung inflammation as demonstrated by lung index, interleukin-6 (IL-6), and interleukin-1β (IL-1β) levels, as well as the pulmonary pathological change. The lung metabolomics coupled with molecular docking showed that FF could alleviate lung inflammation by regulating lysophosphatidylcholine biosynthesis through the caffeoyl quinic acids distributed in the lung tissue. In addition, fecal metabolome coupled with 16S rRNA gene sequencing showed that FF could regulate the tryptophan metabolism by regulating the intestinal flora disorders. This study provided new insights of FF to relieve CS-induced pulmonary inflammation with the multimechanism.

The high mortality rate of chronic heart failure (CHF) makes it a primary battlefield in the field of cardiovascular diseases. Qiangxin Lishui Prescription (QLP) is a traditional Chinese medicine (TCM) prescription used clinically for treating CHF, but its underlying mechanism remains unclear. This study integrated plasma metabolomics, network pharmacology, and experimental validation to reveal the pharmacological effects of QLP and its potential mechanism of anti-CHF. Using ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS/MS), 119 absorbed prototype compounds of QLP were identified from rat plasma. By applying network pharmacology and molecular docking techniques, a QLP absorption components–target–CHF network was constructed. The IL6/JAK/STAT3 signaling pathway is likely critical to QLP's therapeutic effect on CHF. A CHF mouse model was established using aortic ligation surgery to investigate the regulation of the IL6/JAK/STAT3 pathway by QLP in CHF mice. Network pharmacology analysis and in vivo experimental data indicate that QLP alleviates myocardial injury and inflammatory response in CHF mice by modulating the IL6/JAK/STAT3 pathway, significantly improving cardiac function. This presents a promising therapeutic strategy for CHF treatment.

Paxalisib is a dual PI3K/mTOR inhibitor, being used in advanced cancer treatment. In this research, we report a validated LC–MS/MS method for quantifying paxalisib from mouse dried blood spot (DBS). We validated the method in-line with the FDA guidelines. Liquid–liquid extraction technique was used to extract paxalisib from the DBS discs. We used a Chromolith RP-18 end cap (100 × 4.6 mm) column and isocratic mobile phase for the chromatographic separation of paxalisib and the internal standard (IS, dasatinib). The flow was 0.80 mL/min. In the optimized chromatographic conditions, the retention of paxalisib and the IS was ~2.13 and 2.06 min, respectively. Each injection total run time was 2.50 min. The MS/MS ion transitions monitored were m/z 383.2 → 309.1 and 488.1 → 410.1 for paxalisib and the IS, respectively. We have used a broad calibration range (1.24–3762 ng/mL) with a determination coefficient (r²) of 0.995. All the validation parameters assessed met the acceptance criteria, and hematocrit had no effect on DBS Paxalisib concentrations. We have used the validated method to derive the intravenous and oral pharmacokinetic parameters by quantifying paxalisib in mouse blood and correlated with mice pharmacokinetic data.

Mobocertinib is a potent selective tyrosine kinase inhibitor approved for the treatment of non–small cell lung cancer with activating EGFR exon 20 insertions. The aim of this study was to develop a procedure for liquid chromatography tandem mass spectrometry (LC-MS/MS) for the determination of mobocertinib and its metabolite desmethyl-mobocertinib in human plasma. The human plasma samples were precipitated with acetonitrile and analyzed using a Waters ACQUITY BEH C₁₈ column coupled to a triple quadrupole mass spectrometer. Separation was executed using the acetonitrile–0.1% formic acid solution with gradient elution, at a flow rate of 0.4 mL/min. Mobocertinib and desmethyl-mobocertinib were monitored by multiple reaction monitoring (MRM) with m/z 586.5  >  72.2 and 572.4  >  473.2, respectively. The procedure demonstrated excellent linearity (r > 0.997) within the concentration range of 0.1–200 ng/mL for both analytes. Precision in relative standard deviation was <  9.37% for mobocertinib and <  12.03% for desmethyl-mobocertinib. Accuracy in relative error was within −7.23% to 9.18% for mobocertinib and −2.78% to 9.87% for desmethyl-mobocertinib. Extraction recovery was >  80% for both analytes. The validated LC-MS/MS method was successfully applied to the pharmacokinetic study of mobocertinib and desmethyl-mobocertinib in healthy human volunteers with K₂EDTA as anticoagulant after a single dose of mobocertinib (160 mg).

The objective of this study is to comprehensively to identify the core pharmacological components and their respective targets of three medicinal fungi Sanghuangs including Sanghuangporus vaninii (SV), Sanghuangporus lonicericola (SL), and Inonotus hispidus (IH). Metabolomics analysis indicated that a total of 495 and 660 differential metabolites were obtained in mycelium and fermentation broth samples among three Sanghuangs, respectively. The network pharmacology analysis showed that 6-[1]-ladderane hexanol, R-nostrenol, candidone, ellagic acid, and quercetin were the overlapping active ingredients of three Sanghuang species for diabetes mellitus, immune system disease, and neoplasm. Certonardosterol A, dalamid, and ethylene brassylate are unique active ingredients in SV, and certonardosterol K, kaempferide, and esculetin are unique active ingredients in SL. Asbestinine, neoandrographolide, isosakuranetin, and daucosterin are unique active ingredients in IH. Accordingly, the common core targets of active ingredients of the three Sanghuangs were ESR1, PIK3CA, and LYN. PRKCA, EGFR, and STAT3 were the unique targets of SV, SL, and IH, respectively. The primary active components and their respective targets, in addition to the component–target interaction of Sanghuangs that have been identified in the present study, provide a foundation for future research on the prevention and treatment of disease using Sanghuangs.

Proteolysis targeting chimera (PROTAC) is emerging as a promising medicinal modality, which has aroused widespread interest among the field of pharmaceutical manufacturing in the recent years. ARV-471 is an orally active PROTAC estrogen receptor degrader against breast cancer, which leads to the ubiquitylation and subsequent degradation of estrogen receptors via the proteasome. In this study, we developed a highly sensitive liquid chromatography tandem mass spectrometry method (LLOQ = 0.5 ng/mL) for the measurement of ARV-471 in rat plasma. The acetonitrile precipitated sample was separated on ACQUITY BEH C₁₈ column using acetonitrile-0.1% formic acid as mobile phased with gradient elution. Multiple reactions monitoring in positive ESI mode was employed for the quantification of ARV-471 (m/z 724.4 → 396.2). The assay showed good linearity over the concentration range of 0.5–1000 ng/mL with correlation coefficient > 0.996. The assay was validated according to FDA guidance, and all the validation parameters were within the predefined acceptance criteria. After validation, the assay was applied to the pharmacokinetic study of ARV-471 in rats. Additionally, the metabolites in rat plasma were identified using liquid chromatography–high resolution mass spectrometry. Four metabolites were identified and characterized. Hydrolysis, glucuronidation and deamination were the main metabolic pathways of ARV-471 in rats.

Enantioseparation and enantiorecognition are crucial in the pharmaceutical analysis of chiral substances, impacting safety, efficacy, and regulatory compliance. Enantioseparation refers to the process of separating enantiomers from a mixture, typically achieved through chromatography techniques like HPLC and SFC. In contrast, enantiorecognition involves the identification of enantiomers based on their interaction with a chiral selector without the need for separation. Recent advancements in these techniques have significantly improved enantioseparation efficiency and resolution. Chiral stationary phases (CSPs) have evolved, offering better selectivity, including hybrid organic–inorganic materials and miniaturization. The use of green solvents has also reduced environmental impact. Non-chromatographic methods, such as circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy, enable enantiorecognition through interactions with polarized light or chiral solvents. However, these methods face challenges, including high costs, limited solvent compatibility, and shorter operational lifespans compared with chromatographic techniques. Recent developments in solvent-tolerant hybrid CSPs aim to address these limitations. This review highlights these innovations, focusing on their relevance to the pharmaceutical industry, pollution control, and quality assurance, and emphasizes the growing importance of these techniques in the production and regulation of chiral drugs.

Ginkgolide B (GB) is the main active ginkgolide in Ginkgo biloba leaves extract. Pharmacological study suggested that GB exhibits protective effect on nervous system impaired and can be used in the treatment of dementia, cerebral insufficiency or related cognitive decline. However, the information on pharmacokinetics of GB in vivo was limited. In this study, a sensitive LC–MS/MS analytical method was developed to accurately determinate the concentration of GB in dog plasma and applied to the pharmacokinetic study of GB in dogs after bolus injection of GB. A portion of 100 μL dog plasma was pretreated by liquid–liquid extraction using 1 mL of ethyl acetate. The GB and IS were separated on a Waters ACQUITY HSS T3 column using acetonitrile and water containing 0.1% formic acid as mobile phase, at a flow rate of 0.4 mL/min. Multiple-reaction monitoring (MRM) mode was used for quantitative analysis of GB and tolbutamide (internal standard) in negative electrospray ionization. GB showed excellent linearity with correlation coefficient >  0.99 over the concentration range of 1–5000 ng/mL. The intra and interday RSD% were less than 7.4%, while the RE% ranged from −6.1% to 9.6%. The mean extraction recovery was >  83.1%. The validated method was further successfully applied to pharmacokinetic study of GB in dog plasma. Dose-proportional pharmacokinetics of GB were observed after bolus injection administration in dogs. This comprehensive study of the pharmacokinetics of GB in dogs will provide useful information for its further development in clinic.

High-intensity focused ultrasound (HIFU) is a noninvasive soft tissue ablation technique, which utilizes ultrasound energy to induce thermal coagulation necrosis in targeted tissues. Whether this high energy causes side effects in vivo, such as the formation of peptide bonds, has not been fully investigated. Glycylglycine is the simplest dipeptide and hence is often used as a model compound for peptide studies. In this study, we developed and validated a sensitive quantification method based on ion-exchange solid-phase extraction, liquid chromatography, and tandem mass spectrometry (SPE-LC-MS/MS) for the analysis of glycylglycine without derivatization, and then used it to evaluate whether HIFU promoted peptide bond formation in aqueous solution (without enzymes) and plasma (with enzymes). The results showed that strong cation exchange SPE significantly reduced the matrix effect and improved the sensitivity of the LC-MS/MS method. No formation of glycylglycine in the aqueous solution or plasma was observed following HIFU irradiation.