Biomedical Chromatography最新文献

The medicinal plant Tribulus terrestris L. (TT) contains antioxidant and pharmacologically active phenolic compounds. However, extraction procedures can greatly affect extract yield, composition and biological activity. This study evaluated how conventional and advanced extraction methods affected TT extracts' phenolic content and antioxidant activity. TT was extracted using maceration, Soxhlet, ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) with ethanol–water solvent systems applied for different extraction times. The extracts were assessed for total phenolic content (TPC), total phenolic acid content (TPAC) and antioxidant capacity through 2,2-diphenyl-1-picrylhydrazyl (DPPH), cupric ion reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) assays. Phenolic acid composition was analysed by HPLC. TPC values ranged between 27.66 and 37.98 mg GAE/g and TPAC between 8.273 and 24.337 mg CAE/g. Antioxidant activity (IC₅₀) varied from 0.1132 to 0.1904 mg/mL, with UAE (80:20, 10 min) demonstrating the highest DPPH activity. CUPRAC and FRAP values extended from 97.462 to 308.769 mg TE/g and 8.917–17.291 mg TE/g, respectively, with UAE consistently yielding the most active extracts. HPLC detected eight phenolic acids, with protocatechuic acid (2.794 mg/g extract) being the most abundant, followed by p-coumaric, chlorogenic and caffeic acids. The extraction method affected the TT extracts' phenolic content and antioxidant capability. The UAE produced the highest phenolic-rich and bioactive extract. To maximise T. terrestris' phytochemical and nutraceutical potential, extraction procedures need to be optimised.

Antimicrobial resistance is one of the most important global problems, and new antibiotic requirements have been emerging as a key point in this issue. In the present work, we focused on the efficiency of two novel promising fluoroquinolone derivatives on resistant Escherichia coli isolates at the molecular level. Their mode of action and adaptation process were evaluated by using proteomics and metabolomics analysis. Proteomics analysis showed that two compounds have an effect mainly on the ribosomal process and energy metabolism. Moreover, we observed compounds that affect various important antimicrobial targets, such as ribosomal subunits, phosphotransacetylase, and chaperone proteins. In metabolomics analysis, we found that compounds altered bacterial metabolism directly. Pathway analysis showed that cofactor biosynthesis and energy metabolism were affected mainly by undertreated groups. Our experiments demonstrated that novel fluoroquinolone derivatives have promising results at the molecular level and results will contribute to further studies.

Topoisomerase inhibitors (TIs) are a cornerstone class of anticancer and antimicrobial agents, yet their accurate quantification in pharmaceutical formulations and biological matrices remains analytically challenging. These difficulties stem from marked structural diversity, poor aqueous solubility, narrow therapeutic windows, very low circulating concentrations, chemical instability, and, for camptothecin derivatives, pH-dependent interconversion between active lactone and inactive carboxylate forms. This review critically evaluates current analytical approaches for determining widely used TIs, including topotecan, irinotecan, etoposide, epirubicin, dexrazoxane, and camptothecin, with emphasis on selectivity, sensitivity, applicability, and robustness in complex matrices. Conventional spectroscopic methods and HPLC with UV or fluorescence detection remain useful for formulation analysis and preliminary screening but often lack sufficient selectivity and sensitivity for biological samples. Electrochemical techniques, especially those employing nanomaterials or molecular recognition elements, offer high sensitivity and low sample consumption; however, their routine application is limited by matrix effects and scarce regulatory validation. In contrast, ultrahigh-performance liquid chromatography coupled with tandem or high-resolution mass spectrometry (UHPLC–MS/MS or UHPLC–HRMS) provides superior selectivity, sub-ng mL⁻¹ sensitivity, and reliable discrimination of parent drugs and metabolites across diverse matrices. Overall, UHPLC–MS-based methods emerge as the current gold standard for TI quantification, supporting clinical, pharmacokinetic, and regulatory applications in modern analytical science.

This study reports the first bioanalytical method for simultaneous estimation of Tramadol HCl and dexketoprofen trometamol in rat plasma using Tapentadol as an internal standard. A validated LC-MS/MS method was developed following USFDA guidelines. Analytes were extracted from plasma by protein precipitation technique using acetonitrile. Chromatographic separation was achieved on a Waters Symmetry Shield RP-18 column (250 × 4.6 mm, 5 μm) with an isocratic mobile phase of acetonitrile and 0.1% formic acid in HPLC grade water (30:70 v/v) at a flow rate of 1.0 mL/min, yielding a 7-min runtime. Detection was performed using electrospray ionization with ion transitions of m/z 222.3398 → 70.0307 for Tapentadol, m/z 264.4351 → 96.4527 for Tramadol HCl, and m/z 376.4239 → 100.4520 for dexketoprofen trometamol. The method showed good accuracy, with mean recoveries of 92.79%–98.15% for Tramadol HCl and 91.78%–98.37% for dexketoprofen trometamol. Excellent linearity was obtained, with r² values of 0.99983 (22.5–900 ng/mL) and 0.99963 (7.5–300 ng/mL), respectively. All validation parameters met acceptable criteria. The method is suitable for evaluating pharmacokinetic parameters that indicate drug efficacy and safety.

Fuzheng pill (FZP) is a traditional Chinese medicine formula with anti-hepatocellular carcinoma (HCC) effect in clinical practice, but its therapeutic mechanism is still unclear. In this study, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method was used to analyze the chemical composition of FZP, and combined with in vivo and in vitro experiments, to explore its potential mechanism of action in the treatment of liver cancer. A total of 236 components were identified in FZP. In vivo experiments revealed that FZP suppressed tumor growth, improved immune function, and reversed liver and kidney injury caused by tumors, as shown by significant increases in the immune organ index, and that the levels of IFN-γ, IL-2, and TNF-α significantly increased after its administration. Serum biochemical indicators of liver and kidney function decreased. FZP inhibited the proliferation of HepG2 cells. Flow cytometry and Western blot results revealed that FZP modulated the expression levels of Bcl-2, Bax, Caspase-3, and cleaved Caspase-3, and promoted cell apoptosis. FZP may promote the apoptosis of tumor cells by activating the endogenous Bax/Bcl-2/cleaved Caspase-3 apoptosis pathway, which is mediated by mitochondria. These findings revealed the potential of FZP in the treatment of HCC and provided more treatment options for patients.

Wine-steaming of Cnidii Fructus is a traditional processing method believed to enhance efficacy, but its mechanistic basis remains unverified. This study integrated HPLC-based chemical profiling with in vivo anti-osteoporotic evaluation in zebrafish to elucidate the effects of processing. Comparative fingerprinting and multicomponent quantification delineated distinct chemical profiles between raw Cnidii Fructus (RCF) and wine-steamed Cnidii Fructus (WCF). Multivariate analysis (PCA, OPLS-DA) pinpointed six differential coumarins. Quantitative analysis confirmed a marked decrease in imperatorin but increases in osthole, alloimperatorin, and xanthotoxol after processing. Critically, the WCF extract demonstrated superior efficacy over RCF in promoting bone mineralization and enhancing alkaline phosphatase (ALP) activity in an osteoporotic zebrafish model. The differential compounds counteracted dexamethasone-induced impairment, with xanthotoxol showing the most potent osteogenic activity, followed by alloimperatorin, diosmin, osthole, and imperatorin. These findings establish that wine-steaming potentiates the anti-osteoporotic activity of WCF by modulating its coumarin composition, providing a chemical and pharmacological rationale for its traditional use and quality control.

This study investigated the chemical profile and therapeutic mechanisms of the Yishen Ningxin Formula (YSNXF) in treating hypertension comorbid with insomnia through an integrated approach involving UPLC-QE-MS/MS, network pharmacology, and molecular docking. Chemical profiling identified a total of 870 components within YSNXF, with 13 compounds confirmed as blood-entering components in rat plasma, corresponding to 133 potential targets. Intersection analysis with disease-related targets revealed 46 core targets, notably ACE, HTR2A, HTR2B, and HTR1A. Network pharmacology and enrichment analyses indicated that key bioactive components-including hirsutine, geissoschizine methyl ether, hypaphorine, and N-acetylleucine-exert their effects via critical pathways such as the renin–angiotensin system, neuroactive ligand–receptor interactions, and serotonergic synapses. Molecular docking further validated the strong binding affinities of geissoschizine methyl ether and hypaphorine to the identified core targets. These findings suggest that YSNXF alleviates hypertension and insomnia through a multi-component, multi-target, and multi-pathway synergistic mechanism, highlighting geissoschizine methyl ether and hypaphorine as pivotal active constituents for further pharmacological validation.

Hypericum species have been used for medicinal purposes since ancient times because of their important pharmacological properties. Hypericum triquetrifolium Turra belongs to the Hypericaceae family and is native to Eastern Europe and the Mediterranean region. In this study, the essential oil and aroma contents of H. triquetrifolium collected from 21 different localities in the eastern region of Turkey were determined via GC–MS/FID. In addition, the antioxidant and enzyme inhibition activities (cholinesterase, tyrosinase and urease) of the essential oils and major components were determined. Finally, the essential oil and aroma contents were evaluated chemically. The major components of H. triquetrifolium collected from 21 different locations were generally determined to be octane, 2-methyl (1.35%–25.18%), α-pinene (1.32%–19.1%), nonane, 3-methyl (2.43%–24.59%) and caryophyllene (3.79%–30.78%). When the aroma content was examined, octane, 2-methyl (14.22%–44%), nonane (6.37%–18.29%), nonane, 3-methyl (7.62–23.12%) and decane, 2-methyl (3.46%–22.74%) were generally identified as the major components. Overall, all samples were found to have moderate antioxidant capacity. In the AChE enzyme inhibition test, at a concentration of 100 μg/mL, samples S-19, S-9 and S-11 (inhibition %: 87.61 ± 0.63; 84.29 ± 1.51; 82.28 ± 0.53, respectively), and in the BChE enzyme inhibition test, the S-1, S-4 and S-6 samples (88.76 ± 0.79; 91.29 ± 1.58; 93.15 ± 2.16, respectively) were found to be more active than the reference galantamine. In the tyrosinase enzyme inhibition test, samples S-5 and S-13 (inhibition %: 57.09 ± 2.53 and 66.86 ± 0.50, respectively) were found to have greater activity than the other samples. A total of 21 H. triquetrifolium samples collected from different regions of Diyarbakır, Mardin, Elazığ, Siirt, Batman and Şanlıurfa Provinces were analysed via principal component analysis (PCA) and cluster analysis (CA) techniques on the basis of volatile and aroma components. PCA revealed that the dominant volatile components in the samples formed three groups, and regional differences had no significant effect on the samples. The first two principal components explained 59.2% of the variance. The antioxidant capacities of essential oils, their high contents of compounds such as α-pinene and caryophyllene, and their strong acetyl and butyrylcholinesterase enzyme inhibition activities are considered to have potential for use in the food, cosmetics and pharmaceutical industries.

Coccidiosis, caused by Eimeria spp., is a major parasitic disease affecting livestock and laboratory animals, with increasing drug resistance driving the search for natural alternatives. Tirmania nivea, an edible desert truffle rich in bioactive compounds, remains underexplored for this purpose. This study assessed in vitro anticoccidial activity of T. nivea methanolic extract (TNE) against oocyst sporulation and investigated molecular interactions of its major constituents with two key Eimeria enzymes, L-lactate dehydrogenase (LDH) and calcium-dependent protein kinase 1 (CDPK1), using molecular docking. TNE was prepared by methanol–water extraction of fruiting bodies and characterized by GC–MS, identifying 15 major compounds. In vitro assays showed a significant, dose-dependent inhibition of oocyst sporulation, analyzed by one-way ANOVA with post hoc multiple comparisons (p < 0.05). The highest concentration (300 mg/mL) achieved 94.12% and 92.86% inhibition at 72 and 96 h, respectively, exceeding the reference drug amprolium. Docking analyses revealed strong binding affinities of lupeol and estradiol to LDH and CDPK1 (−7.31 to −7.46 and −8.23 to −8.96 kcal/mol, respectively), supported by hydrogen bonding and hydrophobic interactions. ADMET predictions indicated favorable pharmacokinetic properties and low toxicity. These findings support T. nivea as a promising natural anticoccidial candidate targeting essential metabolic and signaling pathways.

In this study, phenolic acid content of bee pollen ethanol extract was investigated using TLC and LC-MS-MS and HPLC-RID methods, and vanillic acid, caffeic acid, gallic acid, and ellagic acid were identified. Antimicrobial activity of bee pollen ethanol extract mixture was screened against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) species by using TLC bioautography, minimum inhibitory concentrations (MIC), and disc diffusion methods. Pollen extract showed antibacterial activity against S. aureus and E. coli; disc diffusion test results showed 16 and 10 mm inhibition zone at 6000 μg/mL concentration. Caffeic acid and gallic acid showed inhibitory action against both S. aureus and E. coli in TLC bioautography studies. Glucose (3.78 ± 0.14 mg/mL), fructose (6.56 ± 0.17 mg/mL), and sucrose (0.13 ± 0.02 mg/mL) were detected as main carbohydrates in pollen extract. Extract did not show significant cytotoxic effect at all tested concentrations on HEK-293 cells.