Biomedical Chromatography最新文献

Compound Nanxing Zhitong Plaster (CNZP) is a topical formulation widely used for rheumatoid arthritis (RA), yet its pharmacodynamic material basis is not fully understood. We established a strategy integrating chemical analysis, network pharmacology, and multi-compartment transdermal pharmacokinetics. Comprehensive chemical profiling by UPLC-Q-TOF-MS identified 100 constituents. Network pharmacology screened 15 core bioactive components. A validated UPLC-QqQ-MS/MS method was developed to simultaneously quantify eight components in the stratum corneum (SC) and viable epidermis plus dermis (VD) and four systemically absorbed components in plasma after transdermal administration, elucidating the formulation's in vivo behavior. Pharmacokinetics revealed structure-dependent transdermal patterns: Phenylpropanoids and phthalides showed rapid absorption and elimination, while organic acids, alkaloids, and flavonoids exhibited reservoir effects and slow release in the SC, contributing to a “fast-slow synergistic” mechanism. Given its high transdermal permeability, notable local and systemic exposure, and established anti-inflammatory evidence, ferulic acid is proposed as a potential quality marker for CNZP. This study provides evidence for the in vivo behavior of topical Chinese medicines and suggests methods for quality control and active substance research.

Massa Medicata Fermentata (MMF), a clinically important fermented traditional Chinese medicine for treating gastrointestinal and metabolic disorders, struggles with quality control due to insufficient reliable reference materials. Because phenolic compounds are key ingredients of MMF, this study investigated how fermentation reshapes the phenolic profile of MMF and enhances its bioactivity. Using UPLC-ESI-MS/MS-based metabolomics, we identified 119 phenolic compounds (PCs), 66 of which were previously unreported in MMF. Comparative analysis showed that fermentation significantly remodeled the PCs composition, notably enriching free phenolic acids, methoxylated flavonoids, and flavonoid C-glycosides compared to the unfermented precursor (BMMF). Consistent with these changes, MMF exhibited significantly stronger antioxidant and anti-inflammatory activities than BMMF (p < 0.01). Correlation analysis revealed strong correlations (∣r∣ > 0.8) between specific PCs and enhanced bioactivities. Our work not only clarifies the molecular basis for fermentation-induced efficacy enhancement but also proposes evidence-based phenolic markers to support the quality standardization of MMF.

Lipids are a diverse class of molecules that mediate numerous structural and functional properties of cells, including trafficking, regulation of membrane proteins, and subcellular compartmentalization. They are the main constituents of biological membranes and mediate transmembrane signaling and structural dynamics. The chemical and structural complexity of lipids makes analysis using a single experimental approach challenging. As such, multiple techniques are used to extract, separate, detect, and quantify lipids, which depend on the specific lipid species and the model being studied. Lipidomics can advance our understanding of the biochemical mechanisms by which lipid-lipid and/or lipid-protein interactions mediate cell growth and death, as well as disease, and provide valuable information for the diagnosis and prognosis of diseases. This review provides an overview of essential methods for the examination of lipids, including extraction methods, chromatographic techniques, and approaches for mass spectrometric analysis. It presents the advantages and disadvantages of commonly used extraction approaches, separation techniques, and mass spectrometry analysis. It emphasizes the need for further standardization of protocols to allow integration of data derived from different platforms. Finally, it discusses the existing opportunities in the field, especially with regard to mass spectrometry imaging and single cell analysis.

Chloroxylon swietenia is a medicinally important tree with documented traditional therapeutic applications. Despite its ethnomedicinal significance, comprehensive phytochemical characterisation and bioactivity evaluation remain limited. Dried leaf powder was subjected to Soxhlet extraction using different solvents with increasing polarity. Phytochemical screening was conducted using standard protocols. Total phenolic and flavonoid contents were determined using Folin–Ciocalteu and aluminium chloride methods, respectively. Total antioxidant capacity was assessed through phosphomolybdenum assay. Functional groups were identified using FTIR spectroscopy, and bioactive compounds were characterised using GC–MS. Phytochemical screening revealed that methanol and acetone extracts contained the highest diversity of secondary metabolites, including alkaloids, flavonoids, phenolics, saponins and tannins. Total phenolic content ranged from 3.387 ± 0.528 to 143.455 ± 1.181 mg/g GAE, with methanol extract showing the highest concentration. Flavonoid content ranged from 10.36 ± 0.468 to 109.85 ± 3.306 mg/g QE. Except aqueous extract, other extracts showed significant antioxidant activity in phosphomolybdenum assay. FT-IR analysis confirmed the presence of diverse functional groups. Through GC–MS analysis, we identified Bioactive compounds, Chalepensin, Chalepin, Lupeol and Isopimpinellin, which are the major compounds with established therapeutic properties. This study demonstrates that Chloroxylon swietenia serves as an exceptional reservoir of therapeutically valuable bioactive compounds, establishing its significance in contemporary pharmaceutical research.

Treosulfan is a bifunctional alkylating agent widely used as a conditioning drug in haematopoietic stem cell transplantation. In this study, a robust reverse-phase UPLC method was developed and validated for the quantitative determination of treosulfan and its related substances. The optimised method employed a Waters Acquity UPLC BEH shield RP-18 (50 × 1.0 mm, 1.7 μm) column with an isocratic mobile phase of acetonitrile and ammonium acetate (pH 2.5)/formic acid in a 40:60 ratio, a flow rate of 0.2 mL/min, injection volume of 5 μL and detection at 239 nm. Validation, performed according to ICH Q2 (R2), confirmed excellent system suitability, specificity, precision (%RSD < 1%), linearity (r² > 0.999), accuracy (recoveries 99%–101%) and sensitivity, with LODs and LOQs meeting acceptable criteria. Forced degradation studies under acid, alkali, oxidative, reductive, photolytic, hydrolytic and thermal conditions confirmed the method's stability-indicating capability, with all purity angles below their respective purity thresholds and acceptable mass balance. LC–MS/MS analysis further enabled structural characterisation of major degradation products. Overall, the developed RP-UPLC method is selective, reproducible and suitable for routine quality control and stability evaluation of treosulfan in bulk drugs.

The aim of this study was to compare blood metabolic profiles between neonates born to propylthiouracil (PTU)-treated hyperthyroid mothers and those born to healthy mothers and to evaluate potential impacts on fetal health despite maternal treatment. Blood metabolites from 31 neonates exposed to PTU and 36 healthy controls were analyzed using liquid chromatography–tandem mass spectrometry (LC–MS/MS) at the Sixth Affiliated Hospital, Sun Yat-sen University, and the Maternal and Child Health Care Hospital of Huadu. Metabolite differences were assessed, and enriched pathway analysis was performed to identify significantly altered metabolic pathways. Neonates exposed to PTU exhibited distinct metabolic profiles compared with controls, characterized by significantly reduced levels of histidine and aspartic acid. These differential metabolites were primarily enriched in ammonia recycling as well as glycine and serine metabolism pathways—closely associated with protein and nucleotide biosynthesis. These findings indicate that prenatal exposure to PTU in hyperthyroid mothers leads to characteristic alterations in neonatal blood metabolic profiles, suggesting the need for close monitoring of growth and developmental outcomes in this population.

Interference is a significant source of laboratory errors that can cause erroneous measurement results. In clinical practice, all the measured results should exclude the influence of interference. In this study, the ion exchange high-performance liquid chromatography of hemoglobin A1c (HbA1c) was evaluated via three brands of HbA1c analyzers, and 12 endogenous and 24 exogenous substances were added to hemoglobin solutions with different HbA1c levels to simulate the corresponding clinical samples. After the potential interfering substances were screened via interference screening testing, the interference concentration thresholds of each potential interfering substance were determined on the basis of dose–response experiment. When these substances exceed the interference concentration thresholds, elevated HbA1c values are caused by endogenous cholic acid and low pH, whereas exogenous ethanol increases HbA1c measurements in samples with low values. However, HbA1c measurements decreased in samples with high values. Although there are some differences in the thresholds of interferent concentrations between different brands of analyzers, the differences are not significant. On the basis of the detailed interference concentration thresholds and interference substances, this evidence can be provided to clinical laboratory experts and help them interpret and determine the testing results of clinical laboratories.