Biomedical Chromatography最新文献

We aim to establish the fingerprint profile of Er Miao San (EMS) that aims to accurately evaluate the quality of EMS samples and to identify quality markers for different batches of EMS and explore its related targets and pathways for the treatment of rheumatoid arthritis. HPLC was employed with an Agilent C18 column. Cluster analysis and principal component analysis were conducted by using SPSS and SIMCA software. Network pharmacology techniques were utilized to explore the potential targets and pathways of EMS in the treatment of rheumatoid arthritis. The HPLC fingerprint profile of EMS was established, featuring a total of 15 common peaks, among which nine were identified. The 10 batches of medicinal materials could be clustered into three categories. With VIP > 1 as the criterion, four index differential components were screened out. These intersections were imported into the STRING database and Cytoscape software, resulting in 228 intersection targets. GO and KEGG enrichment analyses were performed on the related targets and pathways. The established method can effectively analyze the quality differences of EMS from different origins. The screened targets and pathways are of great significance for analyzing the differences in the treatment of rheumatoid arthritis by different batches of EMS.

Dalbergia hancei Benth (D. hancei) is a plant belonging to the Fabaceae family. It has analgesic and anti-inflammatory effects and is used by the Zhuang people to relieve pain. However, the mechanism underlying its analgesic effects remains unclear. This study investigates the analgesic effect of D. hancei based on metabolomics to explain the mechanism of its analgesia from a metabolomics perspective. The analgesic effect was evaluated through the acetic acid-induced writhing test and hot plate test. Three treatment groups received different dosages of D. hancei (0.91 g/kg, 3.64 g/kg, 7.28 g/kg). Its analgesic mechanism was investigated using analgesic behavioral tests and metabolomics. The results of analgesic behavioral experiments showed that all dose groups of D. hancei could relieve pain. A total of eight differential metabolites were identified in the metabolomics results. These biomarkers are associated with five metabolic pathways. Following treatment with D. hancei, eight differential metabolites were identified as regulated, primarily affecting amino acid metabolism, pantothenate and CoA biosynthesis, and steroid hormone biosynthesis. This study revealed the mechanism of analgesia from a metabolomic perspective to provide a basis for screening TCM drugs in pain treatment.

The precise separation of enantiomers is crucial in the pharmaceutical industry for the production of single enantiomer drugs. In this study, a novel chiral composite membrane was developed by embedding L-alanine glutaraldehyde condensation product (L-AGCP) onto a polysulfone (PSf) matrix for enantioselective separation. The PSf-L-AGCP membranes were systematically characterized using spectroscopic techniques to confirm the successful incorporation of L-AGCP and to evaluate the structural properties. Permeation studies using DL-alanine demonstrated that the membrane achieved an enantiomeric excess (ee) of 94% for D-alanine, with a flux of 68.1 mmol·m⁻²·h⁻¹ under optimal conditions of 4 bar of transmembrane pressure and 25 mL·min⁻¹ flow rate. The enantioselective performance of the membrane was influenced by varying in feed concentration, transmembrane pressure, and flow rate, achieving the highest ee at lower feed concentrations and moderate pressures. Computational study using density functional theory (DFT) revealed a significant interaction energy difference between L-AGCP and the alanine enantiomers, with values of −32.63 kJ·mol⁻¹ for L-alanine and −16.94 kJ·mol⁻¹ for D-alanine, explaining the selective retention of L-alanine on the membrane. This study demonstrates the potential of L-AGCP embedded PSf membranes in overcoming the permeability-selectivity trade-off in enantioselective separations, offering a promising avenue for scalable, efficient chiral separations in pharmaceutical applications.

Pectolinarin is a flavonoid compound known for its wound-healing properties, including anti-inflammatory and antibacterial effects. In this study, we employed UPLC-MS/MS to quantify pectolinarin in rat plasma and investigate its pharmacokinetics. Plasma samples were processed using an acetonitrile precipitation method. Chromatographic separation was performed on a UPLC BEH column with a gradient mobile phase of acetonitrile-water (containing 0.1% formic acid). Detection was carried out using electrospray ionization (ESI) tandem mass spectrometry in multiple reaction monitoring (MRM) mode with positive ionization, targeting transitions of m/z 623.3 → 315.3 for pectolinarin and m/z 370.5 → 125.0 for the IS. The results demonstrated that pectolinarin exhibited acceptable linearity in rat plasma within the concentration range of 1.2 to 2300 ng/mL (r > 0.995). The intraday and interday precision, expressed as relative standard deviation (RSD), was below 9.2%. Accuracy ranged from 97.3% to 108.3%, with average recovery exceeding 94.7%. The matrix effect was between 97.8% and 105.3%. The method was successfully applied to evaluate the pharmacokinetics of pectolinarin in rats following both oral and intravenous administration. The absolute bioavailability of pectolinarin in rats was determined to be 0.28%.

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.