Biomedical Chromatography最新文献

A simple, sensitive and efficient ultra performance liquid chromatography–mass spectrometry (UPLC–MS/MS) method for simultaneous quantitative determination of olmesartan and hydrochlorothiazide in human plasma has been developed and validated. Using olmesartan-d₄ and hydrochlorothiazide-¹³C-d₂ as stable isotope-labeled internal standard (SIL-IS). Plasma samples were processed by protein precipitation. Separated on a ZORBAX Eclipse XDB-Phenyl column (150 mm × 4.6 mm, 5 μm) using a gradient elution with mobile phases methanol and 5 mM ammonium acetate. Detected by electrospray ionization (ESI) in negative ion mode of multiple reaction monitoring (MRM), mass transition ion pairs were m/z 445.1 → 149.0 for olmesartan, m/z 295.9 → 204.9 for hydrochlorothiazide, m/z 449.1 → 149.1 for olmesartan-d₄, and m/z 301.0 → 272.0 for hydrochlorothiazide-¹³C-d₂. Linear ranges were 10–2000 ng/mL for olmesartan and 1.5–300 ng/mL for hydrochlorothiazide. Mean recoveries of olmesartan, hydrochlorothiazide, olmesartan-d₄, and hydrochlorothiazide-¹³C-d₂ were 91.31%, 99.34%, 92.62%, and 98.61%, respectively. Our method was well validated in selectivity, carryover, lower limit of quantification (LLOQ), calibration curve, accuracy, precision, dilution effect, matrix effect (normal, hyperlipidemic, and hemolyzed matrices), stability, recovery, and IRS. It was successfully applied in a bioequivalence study of olmesartan medoxomil and hydrochlorothiazide tablets (20/12.5 mg) in healthy Chinese volunteers.

An intravenous drug called sugammadex is used to counteract the adverse reactions of the relaxation medications rocuronium and vecuronium, which are given during operations to provide full anesthesia. The principal objective of the presented study is developing and validating a complete, innovative, fast, isocratic, and green shelf life liquid chromatography methodology, Sugammadex sodium levels in injectable therapies can now be measured. A number of tests were performed on sugammadex sodium in accordance with ICH guidelines. Sugammadex sodium and its related degradants were effectively separated via isocratic separation. At ambient conditions, the Zorbax SB ODS stationary phase was used to quantify and validate the drug substance. The isocratic mobile phase runs at a flow rate of 1.0 mL/min. It includes the eluting phase that comprises of 7 mL/L aqueous solution of triethylamine adjusted at pH 3.0 with phosphoric acid: ethanol (750:250) (V:V). The positive environmental impacts of the LC technique are demonstrated by the combined AES grade of 87, CACI rating of 78, AGSA rating of 75, MoGAPI rating of 80, BAGI rating of 82.5, and final whiteness rating of 96.7.

Renshen Zaizao Wan (RSZZW), a traditional Chinese patent drug originated in Qing dynasty, has been used clinically in China to treat stroke. It consists of 65 different types of plant-, animal-, and mineral-derived Chinese medicinal herbs. However, current quality control of RSZZW mainly relies on a thin-layer chromatography (TLC) method. It is difficult to perform quality assessment for this complicated Chinese patent drug. Therefore, the goal of this study is to simplify the identification process and provide a more comprehensive quality assessment method for RSZZW by developing an integrated strategy based on headspace–gas chromatography–ion mobility spectrometry (HS–GC–IMS) and multivariate statistical analysis. Forty-six batches of commercial RSZZW from six different manufacturers were qualitatively analyzed. A total of 129 volatile signals were detected, of which 85 volatile organic compounds were identified, including 19 esters, 15 ketones, 11 terpenes, 11 alcohols, six aldehydes, and several others. By assessment of the ion mobility global profile, the samples from different manufacturers can be well distinguished by principal component analysis.

Aging decreases the function of various organs and cells, accompanied by the loss of physiological functions. In traditional medicine, Morinda officinalis (MO) and its processed products are often used as kidney-reinforcing drugs. This study set out to evaluate the antiaging efficacy of different components of MO in aging rats and determine whether salt-steaming amplifies the pharmacological activity of MO's most potent components. The results showed that total oligosaccharide partial treatment of MO significantly ameliorated the pathological status of the brain tissue, enhanced spatial memory ability, and improved the expression of various oxidative stress-related factors. In addition, this study found that compared with a high dose of the total oligosaccharide MO component, a high dose of salt-steamed Morinda officinalis (SSMO) significantly improved the pathological status of the brain. Additionally, MO and SSMO reduced apoptosis and inflammation in the aging brain via regulating apoptosis-associated proteins and the SIRT1/NF-κB pathway. UPLC/Q-TOF-MS analysis indicated that the compounds with significant differences in total oligosaccharides between MO and SSMO were 1-kestose (GF2), nystose (GF3), 1F-fructofuranosylnystose (GF4), 1,1,1,1-kestohexose (GF5), and fructo-oligosaccharide DP7 (GF6). These results indicated that the total oligosaccharides of SSMO can show vigorous antiaging effects, providing an experimental basis for exploring the processing of MO.

Yi-Mai compound (YMHJ), a traditional Chinese medicine formulation, has demonstrated therapeutic efficacy against atherosclerosis (AS) in clinical observational studies. To elucidate its underlying pharmacological mechanisms, this investigation employed an integrated approach combining ultra-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (UPLC-ESI-MS/MS), network pharmacology analysis, and molecular docking validation. Through systematic chemical profiling, 2136 constituents were identified in YMHJ, with 33 prioritized as potential bioactive components using network pharmacology-based target prediction. Subsequent multiomics analysis revealed 67 disease-related targets and six hub genes (PPARG, PTGS2, STAT3, TLR4, TNF, and TP53) that exhibited critical regulatory roles in AS pathogenesis. Functional enrichment analysis further indicated that YMHJ modulates key atherogenic processes through regulation of lipid metabolism pathways, RAS signaling cascade, and inflammatory response networks. These findings were corroborated by molecular docking simulations, which confirmed direct binding interactions between the identified compounds and core therapeutic targets. This study provides a comprehensive mechanistic framework for the antiatherosclerotic effects of YMHJ, bridging traditional medicinal knowledge with systems pharmacology insights.

To screen for anti-PMO active components in Cornus officinalis based on ERβ enzyme activity using affinity ultrafiltration and molecular docking techniques.ERβ enzyme and UHPLC-Q-Exactive Orbitrap MS were employed to analyze the anti-PMO activity and predict the components of different concentrations of ethanol extracts of C. officinalis. Molecular docking was used to verify the interaction mechanism between small molecule ligands and ERβ. The anti-PMO activity of the predicted components was further verified using MC3T3-L1 cells. The results showed that the ERβ enzyme can serve as a target enzyme for screening anti-PMO components. The 50% ethanol extract exhibited the best activity. UHPLC-Q-Exactive Orbitrap MS identified and analyzed 17 potential active components. Based on the binding rate and molecular docking results, the following three active components were identified: sweroside, 4-hydroxycinnamic acid, and cornuside. In vitro activity validation confirmed that these components have potential anti-PMO effects. The ERβ enzyme can be used as a potential target enzyme for screening anti-PMO active components in traditional Chinese medicine. The combination of affinity ultrafiltration and molecular docking provides an effective and rapid method for active component screening, offering valuable references for the targeted therapy, mechanism exploration, and quality control of traditional Chinese medicine.

Recently, tilapia fish skin (TFS) has been applied to treat wounds. However the biological compounds in TFS have not been identified yet. Therefore, the objective of this study is to isolate and purify specific hydrophobic bioactive peptides using macroporous resin (XAD 7HP). Recovery rates for total peptides obtained by Bacillus licheniformis alcalase and Aspergillus oryzae protease were (81.26% ± 0.19%) and (80.21% ± 0.18%). Both purified hydrophobic bioactive peptides obtained by protease (BBPP) and alcalase (BBPA) presented high antioxidant capacity (30.4 ± 0.5) and (42.1 ± 0.6) mg trolox/g peptides, respectively, measured by the FRAP method. BBPA and BBPP presented significant antimicrobial activity, with inhibition halo diameters of (10 and 11.3 mm) for Staphylococcus aureus and (12.2 and 13 mm) for Pseudomonas aeruginosa, respectively. BBPA and BBPP showed antinociceptive activity presented by the significant effect to decrease the number of abdominal contortions; BBPA (10 mg/kg) showed greater capacity than indomethacin (20 mg/kg). Liquid chromatography-mass spectrometry (LC-MS) identified the primer sequence of amino acids of BBPA as the following: HYP, ASP, SER, GLU. GLY, HIS, ARG, THR, ALA, PRO, TYR, VAL, LYS, ILE, LEU, and PHE. Therefore, BBPA can be used for different applications in the food and pharmaceutical areas.

Bedaquiline (BDQ) is a diarylquinoline used for the treatment of multidrug-resistant tuberculosis (MDR-TB) with a mechanism of action that inhibits the mycobacterial adenosine triphosphate synthase. There is a limited literature on the pharmacokinetics (PK) of BDQ in MDR-TB patients globally and even less from India. We aimed to develop a rapid, selective, sensitive, and robust high-throughput liquid chromatography–tandem mass spectrometry (LC–MS/MS) method to evaluate the PK of BDQ from the plasma of people with MDR-TB. The analyte was extracted from plasma by protein precipitation. The separation was achieved on a reverse phase C18 column with a mixture of aqueous and organic mobile phase in gradient mode at a flow rate of 0.3 mL/min and detected on a triple quadrupole mass spectrometer. Linearity was obtained between 0.0313 to 4.0 mg/L, and the method was validated following bioanalytical method validation guidelines. BDQ concentrations were determined from plasma samples, and intensive PK testing was performed in samples from 23 patients. The median Cmax attained was 1.59 mg/L with a Tmax around 6 h for most of our patients. This method was successfully developed and validated for a pilot PK study in Indian MDR-TB patients.

To explore the therapeutic mechanisms of Modified Si Miao San (mSMS) in the treatment of gouty arthritis (GA) using a combination of LC–MS/MS analysis, network pharmacology, and experimental validation. The pharmacokinetics of mSMS in rat serum were analyzed by LC–MS/MS, and its major active metabolites were identified. Network pharmacology integrated data from multiple databases (ChEMBL, TTD, YaTCM, and DisGeNET) to investigate the interactions between compounds, targets, and pathways. Experimental validation was performed using HE staining, PCR, Western blot, and serum biochemical assays. UHPLC-QE-MS identified 480 metabolites in the mSMS group, 414 in the drug-containing serum group, and 411 in the control group. Venn analysis revealed 23 unique metabolites in the mSMS serum group. Network pharmacology analysis highlighted key biological processes, including extracellular response to stimuli and steroid metabolic processes. KEGG pathway analysis revealed potential therapeutic targets such as TLR4, NLRP3, and CASP-1. Experimental results showed that mSMS alleviated inflammation in GA rats in a dose-dependent manner. It also reduced SUA, BUN, and CREA levels, downregulated TLR4, P65, NLRP3, and CASP-1 expression, and upregulated ARG-1. mSMS exerts anti-inflammatory and urate-lowering effects in GA by modulating the TLR4/NF-κB/NLRP3 signaling axis, enhancing ARG-1 expression, and promoting uric acid metabolism.