Biomedical Chromatography最新文献

A sensitive, selective, and robust ultra-high-performance liquid chromatography method was developed for the simultaneous quantification of anastrozole and five process-related impurities at trace levels (≤ 0.1%). Traditional univariate optimization was replaced with a systematic chemometric approach using Box–Behnken Design to evaluate critical chromatographic parameters—mobile phase pH (3.0–4.0), column temperature (25°C–35°C), and flow rate (0.20–0.60 mL/min). Response Surface Methodology was employed to model retention behavior, resolution, and peak symmetry, with desirability function optimization identifying the ideal conditions (pH 3.20, 29.3°C, 0.33 mL/min). The method achieved baseline separation on an ACQUITY BEH C18 column (2.1 × 50 mm,  1.7 μm) using 10-mM ammonium formate (pH-adjusted) and acetonitrile as the mobile phase. Validation per International Council for Harmonisation Q2(R1) guidelines confirmed excellent linearity (R² ≥ 0.999), precision (%RSD ≤ 1.5% for ANZ; ≤ 5.3% for impurities), accuracy (recoveries: 94%–101%), and sensitivity (limit of detection: 0.011–0.014 μg/mL; limit of quantitation: 0.020–0.025 μg/mL). This study reports the first validated ultra-high-performance liquid chromatography method enabling trace-level quantification of two newly identified (IMP-1 and IMP-2) and three previously reported process-related impurities of anastrozole in a single run, using a Design of Experiments–based optimization approach.

This metabolomic study aimed to investigate the mechanism of intranasal acupuncture in treating allergic rhinitis (AR) by analyzing local metabolic changes in nasal secretions. Twenty AR patients (receiving 14 consecutive days of intranasal acupuncture) and 20 age- and gender-matched healthy controls were enrolled, with samples analyzed via ultraperformance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-Q-Exactive-HRMS). Baseline comparisons identified 342 differential metabolites between AR patients and healthy controls. Biliverdin, a key intermediate in porphyrin metabolism, was significantly downregulated in AR patients (p < 0.05). After treatment, AR patients showed marked symptomatic improvement, as evidenced by reduced Total Nasal Symptom Score (TNSS), Visual Analog Scale (VAS), and Total Nonnasal Symptom Score (TNNSS), along with a significant rebound in biliverdin levels (p < 0.05). Metabolic pathway analysis revealed the porphyrin metabolic pathway was significantly enriched in both the comparison between AR patients and healthy controls and the comparison between posttreatment and pretreatment AR patients. These findings suggest intranasal acupuncture alleviates AR by regulating the abnormal expression of biliverdin in porphyrin metabolism and activating heme oxygenase-1 (HO-1)-mediated antioxidant and anti-inflammatory responses, highlighting biliverdin as a potential biomarker for AR diagnosis and treatment monitoring.

Doxycycline (DC), a second-generation tetracycline antibiotic, is extensively used in human and veterinary medicine for its broad-spectrum activity, favorable pharmacokinetics, and high bioavailability. This review focuses on analytical methods for detecting and quantifying DC and its metabolites in pharmaceutical, biological, and environmental samples. Spectroscopic, chromatographic, and electroanalytical techniques have been explored, with high-performance liquid chromatography (HPLC) emerging as the most reliable due to its high sensitivity, selectivity, and reproducibility, particularly when coupled with fluorescence or mass spectrometric detectors. Although spectroscopic approaches are simple and economical, they often suffer from matrix interferences. Electroanalytical and voltammetric methods, by contrast, offer rapid and cost-effective alternatives suitable for field applications. Major analytical challenges include DC instability, complex sample matrices, and intricate preparation steps affecting quantification accuracy. Future research should focus on green, miniaturized, and automated analytical platforms that integrate nanomaterials, advanced separation techniques, and artificial intelligence to achieve rapid, accurate, and sustainable monitoring of DC and its metabolites in clinical and environmental settings.

Serum pharmacochemistry was applied to identify the blood-absorbed constituents of dry ginger (DG) and processed ginger (PG). Network pharmacology was employed to construct compound–target–pathway interaction networks, which were further validated through molecular docking to assess the binding affinities between ligands and key targets. Furthermore, a dextran sulfate sodium (DSS)-induced murine model of ulcerative colitis (UC) was established to systematically investigate the multi-target regulatory mechanisms of DG and PG. The processing of DG transforms gingerols into shogaols, thereby modifying its bioactive composition and physiological functions. DG, which is rich in 6-gingerol, mitigates intestinal inflammation through the inhibition of the PI3K-Akt signaling pathway, whereas PG increases the levels of shogaols and zingerone, promoting gut barrier restoration and reducing occult bleeding. Our findings illustrate how processing alters the functional properties of ginger, offering valuable insights for the development of ginger-derived products tailored to specific gastrointestinal health benefits.

Zhi-Zi-Hou-Po Decoction (ZZHPD) is a classic traditional Chinese medicine (TCM) formula for treating depression and anxiety. Considering the disadvantages of decoction, our research group prepared ZZHPD into Zhi-Zi-Hou-Po Granule (ZZHPG). However, the chemical composition and mechanism of ZZHPG for the treatment of depression–anxiety comorbidity remain unclear. In this study, the UHPLC-Q-TOF-MS technology was applied to investigate the chemical constituents in ZZHPG, and network pharmacology was used to reveal the mechanism of ZZHPG. A total of 92 compounds were identified in ZZHPG. The network pharmacological analysis showed that 92 compounds regulated 132 pathways through 90 targets, among which 17 compounds maybe the material basis of ZZHPG for the treatment of depression–anxiety comorbidity, and osthole, obovatol, and naringenin may be the main pharmacodynamic components. Further research indicated that the mechanism of ZZHPG in treating depression–anxiety comorbidity may be related to the inhibition of nervous system disorders, activation of GABA receptors, anti-inflammation, and reduction of oxidative stress. The molecular docking results showed that osthole, obovatol, and naringenin exhibited strong binding affinity with the core targets AKT1, ALB, TNF, EGFR, ESR1, and GSK3B. This research laid foundation for clinical application of ZZHPG and provided new ideas for the study of other TCM prescriptions.

A stability-indicating HPLC method was developed to quantify belzutifan and its related and degradation impurities, and LC–MS/MS technology was used for the characterization of stress-induced degradation products (DPs). Adequate separation was attained on an waters X Bridge C18 column with dimensions 150 × 4.6 mm (3.5 μm) at 30°C, utilizing acetonitrile and 0.1% formic acid in water (60:40) v/v as the mobile phase. This method was set at a flow rate of 0.8 mL/min, and a PDA detector was used for detection. The method demonstrated excellent specificity, accuracy (98.24%–101.01% recovery), precision (%RSD < 2), linearity (R² > 0.999), and sensitivity, with resolution > 2.0. Forced degradation studies showed no degradation under UV and thermal stress but significant degradation under acidic, basic, and oxidative conditions, yielding two acid-induced (DP A/B-1 and A-2), one base-induced (DP A/B-1), and one peroxide-induced (DP O-1) DPs. The degradation pathways of DPS were established by using the mass fragmentation patterns. This method represents stability, is compliant with ICH guidelines, and is well suited for routine quality control and comprehensive stability assessment of belzutifan. A representation of the stress study of belzutifan is shown.

Monothioglycerol predominantly acts as an antioxidant in injectable solutions, inhibiting the oxidation and degradation of medications that are sensitive to oxygen, alongside other formulation ingredients. It scavenges free radicals and ceases the chemical chain reactions that result in instability. A thorough, swift, straightforward, and eco-friendly stability-indicating liquid chromatography (LC) method has been developed and validated, facilitating the measurement of monothioglycerol concentrations in injectable solutions. The antioxidant was studied through several tests in accordance with ICH guidelines, including characteristics such as forced degradation studies. Through isocratic separation, monothioglycerol and its related degradants were effectively resolved. The antioxidant was determined using an end-capped octadecylsilyl (C18) column, and the method was validated under room temperature conditions. The isocratic mobile phase operates at a flow rate of 1.0 mL/min and consists of fixed ratios of filtered water, acetonitrile, and orthophosphoric acid. The overall AES grade of 86, AGREE grade of 0.64, MoGAPI grade of 76, BAGI grade of 75.0, and ultimate whiteness of 95.8 all highlight the ecological advantages of the HPLC method. The current research indicates that the proposed methodology is unique, precise, accurate, stable, and environmentally beneficial at a microscale, making it suitable for routine quality control analysis.

Metoclopramide (MCP) is a commonly used prokinetic and antiemetic medication prescribed for managing gastrointestinal motility disorders, nausea, and vomiting. The risk of adverse reactions, as well as accurate quantification of MCP in pharmaceutical products and biological samples, is essential for quality control, pharmacokinetic assessment, and therapeutic monitoring. Relevant studies for this review were gathered from major scientific databases, including Scopus, Web of Science, ScienceDirect, Google Scholar, and PubMed, restricted to publications written in English. This review provides a critical overview of analytical techniques reported over the last 25 years for MCP determination, focusing on chromatographic, spectroscopic, and electroanalytical methods. Chromatographic techniques—particularly high-performance liquid chromatography (HPLC)—remain the gold standard due to their robustness, precision, and compatibility with diverse sample matrices. Advanced LC–MS/MS systems offer exceptional sensitivity, reaching pg–ng/mL levels, which is particularly valuable in bioanalytical applications. HPLC-UV approaches, though less sensitive, are cost-effective and suitable for routine quality assessment. Spectroscopic methods such as UV–Vis spectroscopy are rapid and affordable but limited by poor selectivity in complex matrices. Electrochemical and voltammetric methods have recently gained attention, offering low cost, speed, and remarkable sensitivity, especially with nanomaterial-modified electrodes. Remaining challenges include MCP instability, matrix interferences, and balancing sensitivity with accessibility.

Cnidii Fructus (CF) is a Chinese herbal medicine with bioactive immune properties and potential applications in treating ulcerative colitis (UC), but its well-defined bioactive constituents are understudied. The current study aimed to investigate CF's anti-UC bioactivity and elucidate its bioactive constituents via chromatographic analysis, in vitro/in vivo tests, and spectrum-effect analysis. Results showed 10 batches of CF inhibited the inflammatory response induced by lipopolysaccharide (LPS)–stimulated RAW 264.7 macrophages. High-performance liquid chromatography (HPLC) fingerprint profiles of the 10 batches were established, with 16 common peaks identified. Subsequently, 15 of these peaks (including eight reference-validated compounds) were characterized by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS). Four anti-UC-related compounds were identified via spectrum-effect relationships, namely, diosmin, xanthotoxol, imperatorin, and osthole. Experimental verification confirmed these as key components potentially linked to alleviating inflammatory responses in UC. This study is the first to establish a spectrum-effect relationship for CF's anti-UC effects, identify its critical active constituents, and provide a scientific basis for CF quality control and UC therapeutic development.

The leaves of Dimocarpus longan Lour. (LYY), a plant belonging to the Sapindaceae family, are a traditional Chinese medicine (TCM) with Guangxi national characteristics. However, its quality standards remain inadequate, and considering the impact of the current standard harvest period on medicinal plants, it is necessary to study how different harvest periods affect the component contents of LYY. In this study, high-performance liquid chromatography (HPLC) combined with chemometrics was employed to analyze the chemical components of LYY across various harvest periods. By comparing 12 batches of LYY fingerprints, 8 compounds were identified. The results of similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and weighted technique for order preference by similarity to ideal solution (TOPSIS) revealed differences in LYY samples from different harvest periods, with the samples of January and February being of better quality, finding that gallic acid, quercitrin, and quercetin might be potential markers for these compositional variations. Quantitative analysis demonstrated that the contents of different compounds varied with the harvest period. In conclusion, the established method can be applied to comprehensively evaluate the LYY quality, providing a scientific basis for its reasonable harvest.