Biomedical Chromatography最新文献

Danatinib, a brand-new compound synthesized in our laboratory for the treatment of acute myeloid leukemia (AML), demonstrates remarkable antitumor activity. However, the pharmacokinetic profile of Danatinib in mice was short with its half-life was only 0.476 h. To address this issue and optimize its therapeutic efficacy, this study investigated the metabolic profiles of Danatinib in mice, both in vitro and in vivo, using ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) technology. In the in vitro study, Danatinib was analyzed using mouse liver microsomes, resulting in the identification of eight metabolites. In the in vivo study, Danatinib was administered orally to mice at 20 mg/kg; the samples of plasma, bile, feces, and urine were collected and analyzed, leading to the identification of 34 metabolites. The results showed that those metabolites were formed through various metabolic reactions including hydroxylation, carboxylation, acetylation, hydrogenation, and glucuronidation. This study provides a systematic investigation of the metabolism of Danatinib, offering valuable information for further structural modification to improve its pharmacokinetic profiles.

Tanreqing injection (TRQI), a well-known traditional Chinese medicine, has a marked curative effect on lower respiratory tract infections. A strategy using ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC–Q–TOF–MS^E) for rapid identification of metabolites from TRQI was proposed by UNIFI informatics platform combined with multiple data processing techniques. Target prediction was then performed based on the original compounds in vivo, and a network between the active compounds and common targets was established using Cytoscape v3.9.0. As a result, 64 original compounds were characterized in TRQI, and 54 original compounds and 76 metabolites of TRQI were detected in human plasma and urine, two of which (M28 and M45) were novel metabolites. A novel metabolic pathway for lonicerin was identified. The compound-target-pathway network identified 22 target genes and 20 signaling pathways that were linked to these mechanisms. The key mechanism is related to the JAK-STAT signaling pathway and PI3K-Akt signaling pathway. The bioactive ingredients and mechanisms of action of TQRI against lower respiratory tract infections based on original compounds in vivo were explored through network pharmacology and molecular docking. This is the first study in which the mechanism of action of TRQI in humans has been clarified.

To investigate the material basis for the increased efficacy of Anemarrhenae Rhizoma (AR) after salt processing, this study employed UPLC-Q-TOF-MS/MS technology to analyze metabolites in both AR and salt-processed AR (SAR) extracts, as well as in serum samples treated with these extracts. The results identified 54 chemical components in AR and SAR under both positive and negative ion modes, with 21 prototype components detected in the bloodstream. A comparative analysis of serum absorption levels revealed higher concentrations of neomangiferin, mangiferin, and other chemical constituents in the SAR group than in the AR group. Additionally, 29 differential metabolites were identified, with 12 unique to AR and 17 to SAR. These findings highlight significant alterations in the chemical composition of AR after salt processing and provide a scientific basis for understanding the enhanced efficacy of SAR. This study offers valuable insights for further research into the clinical efficacy of SAR and its applications.

A precise and stability-indicating high-performance liquid chromatographic (HPLC) method was developed and validated for the simultaneous estimation of lobeglitazone sulfate (0.5 mg) and metformin hydrochloride (500 mg) in pharmaceutical formulations. This method effectively addresses the analytical challenge posed by the significantly different dosage strengths and contrasting polarities of lobeglitazone (water insoluble) and metformin (water soluble). Utilizing a Phenomenex Strong Cation Exchange (250 mm × 4.6 mm, 10 μm) column, the method employed a mobile phase of ammonium dihydrogen phosphate (pH 3.0) and a premix of methanol and acetonitrile (80:20), delivered at a flow rate of 1.0 mL/min. The PDA detector was set at 251 nm to monitor eluents, and the column temperature was maintained at 30°C for optimal separation. The method was validated as per ICH guidelines, covering specificity, linearity, accuracy, precision, LOD, LOQ, solution stability, and robustness. Forced degradation studies under various stress conditions (acidic, basic, oxidative, photolytic, and thermal) revealed no significant degradation products. The method was also assessed for greenness using a AGREE tool, achieving a score of 0.68, demonstrating its eco-friendly nature. Overall, this method is suitable for the routine analysis of these drugs in pharmaceutical formulations.

Clozapine is an antipsychotic medication primarily used to treat severe schizophrenia symptoms. This research aimed to develop and validate a reliable, rapid, and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method for measuring organic impurities in clozapine drug substances and products. The chromatographic separation was achieved using an Agilent Poroshell 120 EC-C₁₈ column with gradient elution, using mobile phases consisting of acetonitrile, methanol, pH 2.4 phosphate buffer, and ethanol. The method operated at a flow rate of 1.0 mL/min, with a 10-μL injection volume, a column temperature of 35°C, and detection at 257 nm. Validation was performed according to ICH Q2(R2) guidelines and USP <1225> general chapter. The method demonstrated optimal resolution between adjacent peaks, with values greater than 1.5, and provided accurate estimations free from interference. Recovery and regression values ranged from 80.0% to 120.0%, with R² values exceeding 0.9995. The quantification range spanned from the limit of quantification to 150% of the specification level. The stability-indicating capability of the RP-HPLC method was confirmed through forced degradation studies. This method has been successfully implemented in a quality control laboratory for real-time analysis of clozapine drug substances and products.

Studies reporting on the analysis of free plasma (platelet-poor plasma, PPP) serotonin (5-hydroxytryptamine, 5-HT) in plasma obtained from healthy humans have been systematically reviewed. The review covered the period from 2010 to July 2024 and is a follow-up of a similar review published in 2011 which found that nearly all published reported of PPP 5-HT were clearly and markedly erroneously high. This problem has persisted unabated with nearly all retrieved 47 reports from the past 14 years also apparently being erroneously high. Possible causes and consequences of the problem are discussed along with potential approaches to improving the analysis and reporting of plasma 5-HT. Most of the errors appear to arise from pre-analytical problems that occur during the preparation of PPP due to residual platelets and/or due to the release of platelet 5-HT. The large number of fields interested in plasma 5-HT and the disparate publication venues appear to have contributed to a general lack of awareness of the difficulties with analyzing plasma 5-HT. The reporting of erroneous plasma 5-HT values has led to unsupported conclusions about possible alterations in plasma 5-HT and about the role of plasma 5-HT across a wide range of biomedical research areas.

The chemical biology method is a cutting-edge technology that utilizes chemical research methods and ideas to study life and biomedicine at the molecular level. Recently, it has gradually been applied to screening chemical markers in traditional Chinese medicines. This article reviews the principles and applications of common chemical biology methods, including cell membrane chromatography, affinity ultrafiltration, magnetic bead enrichment, capillary electrophoresis, molecular exclusion chromatography, immobilized fusion target affinity chromatography, etc., in traditional Chinese medicines research. Applying chemical biology methods to screen chemical markers in traditional Chinese medicines has the advantages of simple and fast operation, strong specificity of results, good reproducibility, and high sensitivity, which can achieve high-throughput screening.

Sangge Jiangzhi pill, as a traditional Chinese medicine compound preparation, can be used to treat hyperlipidemia. However, the specific material basis and mechanism of action of the Sangge Jiangzhi pill are not clear. In this study, gut microbiota and host metabolism were investigated using 16S rRNA sequencing and metabolomics techniques. The changes in body weight, organ index, serum biochemical levels, and liver pathology demonstrated that the Sangge Jiangzhi pill reduced body weight and attenuated fat accumulation in hyperlipidemic rats. The results of gut microbiota analysis demonstrated that the Sangge Jiangzhi pill significantly enhanced the richness and diversity of gut microbiota in hyperlipidemia rats. Moreover, it regulated the relative abundance of Ligilactobacillus, Limosilactobacillus, Lactobacillus, Blautia, Parabacteroides, Helicobacter, Colidextribacter, Desulfovibrio, Rikenella, and Tuzzerella to protect the intestinal barrier and relieve the oxidative stress reaction. Metabonomics analysis showed that a total of 32 differential metabolites were identified in feces. The results of the correlation analysis revealed a significant association between intestinal microbiota and fecal metabolites. These results suggested that the Sangge Jiangzhi pill could achieve the purpose of treating hyperlipidemia by regulating gut microbiota imbalance and metabolic activity disorder.

Effective enantiomer separation is vital in many important sectors like pharmaceuticals, agrochemicals, food safety, and biomedical imaging, yet conventional methods are costly, slow, and chemical intensive. This has sparked interest in exploring novel materials like chiral lead halide perovskite nanocrystals to address these challenges. This newly emerging chiral material combines the superior properties of traditional halide perovskites with the unique attributes of chirality, resulting in distinct optoelectronic behaviors. This perspective provides a discussion on future research opportunities in the usage of these chiral LHP NCs for enantiomeric recognition and separation. LHPs exhibit extraordinary photophysical properties, easier surface functionalization, range of bonding interactions, and high surface area to volume ratio that can be used for detecting enantiomers. To the best of our knowledge, the use of chiral halide perovskites for the chiral discrimination of enantiomers has been scarcely reported, presenting a novel opportunity to explore their potential in enantiomeric recognition and separation.

In the current study, a simple ultra-high performance liquid chromatography–tandem mass spectrometry method was developed and fully validated for the quantitation of deacetyl asperulosidic acid methyl ester in rat plasma. The plasma sample was precipitated with acetonitrile and then separated on the Waters ACQUITY UPLC HSS T3 column. The mobile phases, water and acetonitrile, were added with 0.1% formic acid. The mass spectrometry detection was performed in negative-ion multiple reaction monitoring. In the range of 1–1000 ng/mL, the linearity meets the requirements with correlation coefficient more than 0.99. The parameters of accuracy, precision, carryover, matrix effect, extraction recovery, stability, and dilution integrity are within accepted ranges. The validated method has been successfully used for pharmacokinetic study of deacetyl asperulosidic acid methyl ester in rats. After oral administration, deacetyl asperulosidic acid methyl ester was quickly absorbed into blood and reached the maximum plasma drug concentration of 4047.49 ng/mL at 2 h. The half-life of deacetyl asperulosidic acid methyl ester is 5.6 h, which suggests that it has a moderate metabolic process. Since the absolute bioavailability of deacetyl asperulosidic acid methyl ester is only 3.74%, its gastrointestinal stability, first-pass effect, and transmembrane properties remain to be studied.