Journal of pharmaceutical and biomedical analysis最新文献_第7页

Integrating 16S rDNA sequencing analysis and targeted metabolomics to explore the mechanism of Xiexin Tang in treating atherosclerosis mice induced by high-fat diet

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116760

Junling Li , Qianru Gao , Hongtao Liu , Songlin Liu , Yanchun Wang , Xiongjie Sun , Junping Zheng , Huabing Yang , Baifei Hu

{"title":"Integrating 16S rDNA sequencing analysis and targeted metabolomics to explore the mechanism of Xiexin Tang in treating atherosclerosis mice induced by high-fat diet","authors":"Junling Li , Qianru Gao , Hongtao Liu , Songlin Liu , Yanchun Wang , Xiongjie Sun , Junping Zheng , Huabing Yang , Baifei Hu","doi":"10.1016/j.jpba.2025.116760","DOIUrl":"10.1016/j.jpba.2025.116760","url":null,"abstract":"<div><div>Xiexin Tang (XXT) is a classic Chinese medicine formula that can be used to treat Atherosclerosis (AS). This study aimed to investigate the mechanism by which XXT regulated AS lipid levels. Firstly, the mixture components of XXT were analyzed by High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Then, the AS model based on Apolipoprotein E knockout (ApoE<sup>-/-</sup>) mice was established. Cytokines related to lipid metabolism and bile acid metabolism were detected by Quantitative Real-time PCR (qRT-PCR). 16S rDNA gene sequencing was performed to analyze differential bacterial populations, and the mechanism of XXT regulation of bile acids affecting lipid metabolism was further explored by targeted metabolomics. Further, antibiotic-treated mice were used to investigate the role of gut microbiota in the anti-AS effect of XXT. The results showed that XXT attenuated the lipid levels and reversed the abnormal elevation of cytokines, such as hepatic lipid metabolism and inflammatory reaction in AS mice. XXT also repaired the gut barrier damage and reversed gut microbiota disorders in AS mice. Furthermore, the metabolic levels of bile acids were reshaped by XXT. Whereas, in the absence of gut microbiota, XXT failed to attenuate lipid levels and inhibit the expression of cytokines related to inflammation and bile acid metabolism in AS mice and failed to play a role in ultimately treating AS. In conclusion, XXT could effectively inhibit the inflammatory reaction and lipid accumulation in AS mice, and this effect was closely related to its remodeling of gut microbiota to regulate bile acid metabolism.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116760"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Illuminating green fluorescent protein: Characterizing tri-peptide fluorescent chromophore, probing reactivity of cysteines, and unveiling site-directed modifications through mass spectrometry

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116771

Jianmin Zhang, Bing Wang

{"title":"Illuminating green fluorescent protein: Characterizing tri-peptide fluorescent chromophore, probing reactivity of cysteines, and unveiling site-directed modifications through mass spectrometry","authors":"Jianmin Zhang, Bing Wang","doi":"10.1016/j.jpba.2025.116771","DOIUrl":"10.1016/j.jpba.2025.116771","url":null,"abstract":"<div><div>Bioconjugation technologies enable covalent attachment of diagnostic or therapeutic effectuators onto biological targets, allowing for the precise delivery of desired drugs to the intended targets with enhanced potency, selectivity, specificity, and prolonged duration of action. As the number of bioconjugation techniques has grown enormously, identification and in-depth characterization of in-process products play a critical role in the development of covalent drug conjugates. This is especially significant in light of the increased complexity of novel biotherapeutics derived from biological matrices. This paper describes liquid chromatography-mass spectrometry (LC-MS/MS)-based studies that have contributed to the development of site-specific genetic incorporation of non-natural amino acids (nnAAs) into proteins. A holistic approach was implemented to characterize a wild type green fluorescent protein (wtGFP) and an enhanced green fluorescent protein (eGFP). By using the wtGFP as a pilot and model system, the reactivity of cysteine residues was investigated under different sample processing conditions, followed by a stability evaluation using intact mass measurement. The subsequent complementary proteolytic peptide mappings were performed to achieve full sequence coverage of the proteins, identification of predominant modifications, and granular details of the fluorescent chromophore. The developed method was successfully applied to isolate the eGFP incorporated with nnAA from cells. This enables the verification of the specific site of nnAA incorporation, and the characterization of complex variants using <em>de novo</em> sequencing techniques. MS studies demonstrated that <em>p</em>-azido-phenylalanine <em>(p</em>AzF) was specifically incorporated into the desired site of eGFP with high efficiency and fidelity.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116771"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The active metabolite hydroxyitraconazole has substantially higher in vivo free fraction and free concentrations compared to itraconazole

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116775

Motoshi Iwao , Ryota Tanaka , Yosuke Suzuki , Ryosuke Tatsuta , Takehiro Hashimoto , Kazufumi Hiramatsu , Hiroki Itoh

{"title":"The active metabolite hydroxyitraconazole has substantially higher in vivo free fraction and free concentrations compared to itraconazole","authors":"Motoshi Iwao , Ryota Tanaka , Yosuke Suzuki , Ryosuke Tatsuta , Takehiro Hashimoto , Kazufumi Hiramatsu , Hiroki Itoh","doi":"10.1016/j.jpba.2025.116775","DOIUrl":"10.1016/j.jpba.2025.116775","url":null,"abstract":"<div><div>Itraconazole (ITCZ) is a triazole antifungal agent that is metabolized to many products. Hydroxyitraconazole (OH-ITCZ) is the major metabolite with antifungal activity comparable to that of ITCZ. Protein-free drug concentration has been reported to be a better biomarker for pharmacodynamics compared with total drug concentration. We developed an assay for quantification of free ITCZ and free OH-ITCZ concentrations using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) with equilibrium dialysis. The assay fulfilled the requirements of the US Food and Drug Administration guideline for assay validation, with lower limit of quantification of 0.025 and 0.25 ng/mL for free ITCZ and OH-ITCZ, respectively, than previous studies. Using this validated assay, we measured plasma free ITCZ and OH-ITCZ concentrations in 18 samples of 11 adult patients who received oral ITCZ between July 2016 and November 2022 at Oita University Hospital, and compared the <em>in vivo</em> percent free fraction and free concentration between the two compounds. Average plasma free concentrations and percent free fraction in 18 samples were, respectively, 0.188 ± 0.123 ng/mL and 0.024 ± 0.016 % for ITCZ, and 1.449 ± 1.017 ng/mL and 0.251 ± 0.109 % for OH-ITCZ, indicating that OH-ITCZ was 8.52-fold higher in percent free fraction and 10.42-fold higher in free concentration compared to ITCZ. Given that OH-ITCZ and ITCZ have similar <em>in vitro</em> antifungal activity, OH-ITCZ may contribute more to <em>in vivo</em> antifungal efficacy than ITCZ, suggesting that monitoring OH-ITCZ would be more beneficial.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116775"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of pH-adjusted NMR methodology for quantitation of caffeoylquinic acid derivatives and evaluation of their antimalignant pleural mesothelioma potential

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116776

Hau Thi Hong Bui , Yoon-Jin Lee , Trung Huy Ngo , Punam Thapa , Yun-Seo Kil , Chang Yeol Lee , Hyukjae Choi , Kyu Joon Lee , Joo-Won Nam

{"title":"Development of pH-adjusted NMR methodology for quantitation of caffeoylquinic acid derivatives and evaluation of their antimalignant pleural mesothelioma potential","authors":"Hau Thi Hong Bui , Yoon-Jin Lee , Trung Huy Ngo , Punam Thapa , Yun-Seo Kil , Chang Yeol Lee , Hyukjae Choi , Kyu Joon Lee , Joo-Won Nam","doi":"10.1016/j.jpba.2025.116776","DOIUrl":"10.1016/j.jpba.2025.116776","url":null,"abstract":"<div><div>Quantitative analysis of caffeoylquinic acids (CQAs) is challenging due to their structural similarities and susceptibility to isomerization and degradation. To overcome these limitations, a pH-adjusted quantitative <sup>1</sup>H NMR (qHNMR) method is proposed for evaluating different CQAs in the extracts using a MeOD/HEPES-<em>d</em><sub><em>18</em></sub> buffer. This method was applied to measure the concentrations of chlorogenic acid (<strong>1</strong>) and 3,5-di-CQA (<strong>4</strong>) in <em>Cuscuta japonica</em> extract, and of chlorogenic acid (<strong>1</strong>) in green coffee bean extracts. A comparison of quantum mechanics (QM)-based and non-quantum mechanics-based approaches in qHNMR has shown that the application of QM-qHNMR improves the precision and sustainability of quantitative NMR. In addition, <em>C. japonica</em> extract and its isolated CQAs (<strong>1</strong>−<strong>7</strong>) were investigated for anticancer effects against NCI-H2452 cells, showing that methyl di-CQA esters (<strong>5</strong> and <strong>7</strong>) inhibit proliferation and induce apoptosis of NCI-H2452 cells, indicating their potential as therapeutic agents for treating malignant pleural mesothelioma.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116776"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unraveling the cytotoxicity and cellular pharmacokinetic of mPEG5-NH2 polymers by UHPLC-MS/MS

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116767

Yingxia Guo , Meichen Liu , Yajie Cheng , Jiye Tian , Chunpeng Feng , Qingbin Wang , Xuan Zhao , Lei Yin

{"title":"Unraveling the cytotoxicity and cellular pharmacokinetic of mPEG5-NH2 polymers by UHPLC-MS/MS","authors":"Yingxia Guo , Meichen Liu , Yajie Cheng , Jiye Tian , Chunpeng Feng , Qingbin Wang , Xuan Zhao , Lei Yin","doi":"10.1016/j.jpba.2025.116767","DOIUrl":"10.1016/j.jpba.2025.116767","url":null,"abstract":"<div><div>Methoxy-polyethylene glycol amine (mPEG-NH₂), an important pharmaceutical excipient, has been extensively utilized in the field of biomedicine. To advance the development of mPEG-NH₂-related drug delivery systems, it is crucial to understand the safety profile and cellular pharmacokinetic behavior of mPEG-NH₂ polymers. In this study, a straightforward analytical assay based on ultra-high performance liquid chromatography and tandem mass spectrometry (UHPLC-MS/MS) for quantifying mPEG<sub>5</sub>-NH₂ in biological matrices was established and validated. Multiple reaction monitoring (MRM) transitions were selected for quantification of mPEG<sub>5</sub>-NH₂ (mass-to-charge ratio, <em>m/z</em> 252.1 → 87.7) and octaethylene glycol (OH-PEG<sub>8</sub>-OH) (mass-to-charge ratio, <em>m/z</em> 371.2 → 89.2). The UHPLC-MS/MS assay demonstrated excellent linearity within the concentration range of 0.01–10 μg/mL, with an R² value of 0.9996. Both intra-day and inter-day accuracies and precisions of the analytical method were within ± 9.19 %. This analytical assay was successfully applied to study the <em>in vitro</em> cellular toxicity and uptake behavior of mPEG<sub>5</sub>-NH₂ in MCF-7 cells. The results indicate that high doses of mPEG<sub>5</sub>-NH₂ may have potential toxicity to MCF-7 cells.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116767"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143521016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Machine learning enabled protein secondary structure characterization using drop-coating deposition Raman spectroscopy 利用滴涂沉积拉曼光谱进行机器学习，确定蛋白质二级结构特征

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116762

Jeremy Peters , Chunguang Jin , Anna Luczak , Brendon Lyons , Ravi Kalyanaraman

{"title":"Machine learning enabled protein secondary structure characterization using drop-coating deposition Raman spectroscopy","authors":"Jeremy Peters , Chunguang Jin , Anna Luczak , Brendon Lyons , Ravi Kalyanaraman","doi":"10.1016/j.jpba.2025.116762","DOIUrl":"10.1016/j.jpba.2025.116762","url":null,"abstract":"<div><div>Protein structure characterization is critical for therapeutic protein drug development and production. Drop-coating deposition Raman (DCDR) spectroscopy offers rapid and cost-effective acquisition of vibrational spectral data characteristic of protein secondary structures. Amide I region (1600 −1700 cm<sup>−1</sup>) and amide II region (1500–1600 cm<sup>−1</sup>) of DCRD Raman spectra measured for model proteins of varying molecular size and structural distribution were first analyzed by peak fitting for their proportions of six secondary structure motifs: α-helices, 3<sub>10</sub>-helices, β-sheets, turns (β-turns and γ-turns), bends, and random coil. The high spectral resolution and superior signal-to-noise of DCDR spectra made it possible to estimate all six structural motifs at accuracy comparable to X-ray crystallographic measurement. The ease of DCDR measurement was further explored by introducing machine learning algorithm to spectroscopic data analysis. Partial Least Squares (PLS) regression modeling was used as a machine learning tool to predict the protein secondary structural composition from the amide I band of model proteins. Once developed on a training sample set, the PLS model was tested by applying to a sample set that was not used previously for model development. Low prediction errors were achieved at 1.36 %, 0.78 %, 0.42 % 0.41 %, 0.81 %, and 0.52 %, respectively for the six structural component, α-Helix, β-Sheet, 3<sub>10</sub>-helices, random, turns, and bends. The PLS model was further tested on an independent sample set that contains three IgG proteins. The proportion ofα-Helix, β-Sheet, 3<sub>10</sub>-Helix were estimated with an error of 3.1 %, 2.3 % and 2.8 %, respectively.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116762"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143527319","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development and validation of an LC–MS/MS method for pharmacokinetic assessment of a bispecific antibody (YBSW015) targeting SARS-CoV-2

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-25 DOI: 10.1016/j.jpba.2025.116770

Yuan-Yuan Chai , Ling Zhou , Yan Yang , Lin Tan , Ke Ma , Zhi-Jiang Huang

{"title":"Development and validation of an LC–MS/MS method for pharmacokinetic assessment of a bispecific antibody (YBSW015) targeting SARS-CoV-2","authors":"Yuan-Yuan Chai , Ling Zhou , Yan Yang , Lin Tan , Ke Ma , Zhi-Jiang Huang","doi":"10.1016/j.jpba.2025.116770","DOIUrl":"10.1016/j.jpba.2025.116770","url":null,"abstract":"<div><div>YBSW015 is a bispecific antibody that targets severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A pair of high-affinity parental monoclonal antibodies (B38 and H4) were discovered from the sera of convalescent patients, and YBSW015 was developed based on these two parental antibodies. To support pharmacokinetic research, a simple, specific and reproducible liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantifying YBSW015 in human serum. Two signature peptides (SPs) were selected as surrogate analytes for quantification and qualitative detection, respectively. Two stable isotope-labeled synthetic peptides with the same sequence of two SPs were set as internal standards (ISs). Total antibody was precipitated with 1.0 % (w/v) trichloroacetic acid (TCA) in isopropanol (IPA) following by trypsin enzymatic and further purified by solid phase extraction (SPE). The performance of this bioanalytical assay was well evaluated with respect to linearity, accuracy and precision, selectivity, specificity, matrix effect, recovery, and stability. YBSW015 concentrations in human serum were determined over the concentration range of 2.00–1000 μg/mL, and the quantification was not interfered by the matrix components and pre-existing antibodies. This precise and sensitive method was successfully employed in a phase I clinical study.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116770"},"PeriodicalIF":3.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143549087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of oral fluid in urine samples on the analysis of selected erythropoietin receptor agonists and detection of saliva-specific markers for doping control purposes

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-24 DOI: 10.1016/j.jpba.2025.116769

Ann-Marie Garzinsky , Judith Harth , Florine Leipp , Katja Walpurgis , Philipp Reihlen , Andreas Thomas , Mario Thevis

{"title":"Effect of oral fluid in urine samples on the analysis of selected erythropoietin receptor agonists and detection of saliva-specific markers for doping control purposes","authors":"Ann-Marie Garzinsky , Judith Harth , Florine Leipp , Katja Walpurgis , Philipp Reihlen , Andreas Thomas , Mario Thevis","doi":"10.1016/j.jpba.2025.116769","DOIUrl":"10.1016/j.jpba.2025.116769","url":null,"abstract":"<div><div>Due to their performance-enhancing effect, erythropoiesis-stimulating agents (ESAs) are banned at all times by the World Anti-Doping Agency (WADA) in competitive sports. Doping control analyses for such compounds are routinely performed using gel electrophoretic and immunoblotting techniques, and degradation of the analytes can severely impair detection, results evaluation and interpretation. As oral fluid (OF) contains significant amounts of proteases, the question of whether its addition to a doping control urine sample may impede anti-doping analysis needs to be addressed. Intentional tampering attempts are likewise prohibited by WADA and require a detection strategy. It was observed that the addition of OF can indeed lead to impairments of ESA analyses, though the fraction of unidentifiable ESA signals varies depending on several factors, such as the individual composition of the OF, the sex of the OF donor, the time of sampling, the OF volume and the incubation conditions. Overall, 20 % of all generally valid analyses were classified as unidentifiable, 12 % as impaired, and 69 % as identifiable, highlighting the relevance for strategies that allow for the identification of OF in urine. While human salivary α-amylase was found insufficiently reliable as a marker, peptides of salivary proline rich proteins (saPRP) were shown to be both specific for OF and traceable with adequate sensitivity using a newly developed LC-HRMS/MS method. The approach was comprehensively characterized shown to be fit-for-purpose for routine doping controls where tampering attempts with OF are suspected.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116769"},"PeriodicalIF":3.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143509678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification of chemical components and metabolites in rats after oral administration of Epimedium-Astragalus granule pair by liquid chromatography-high resolution mass spectrometry combined with diagnostic fragment ions and mass defect filtering

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-24 DOI: 10.1016/j.jpba.2025.116768

Song Wang , Xinnan Chang , Jing Li , Zuoqiao Shi , Guowen Li

{"title":"Identification of chemical components and metabolites in rats after oral administration of Epimedium-Astragalus granule pair by liquid chromatography-high resolution mass spectrometry combined with diagnostic fragment ions and mass defect filtering","authors":"Song Wang , Xinnan Chang , Jing Li , Zuoqiao Shi , Guowen Li","doi":"10.1016/j.jpba.2025.116768","DOIUrl":"10.1016/j.jpba.2025.116768","url":null,"abstract":"<div><div>Herbal pairs are combinations of two relatively fixed herbs that are frequently used in clinical practice to achieve specific therapeutic effect. Epimedium and Astragalus are frequently used together in clinical settings. However, there is a lack of an in-depth understanding of the active components of these herbs <em>in vivo</em>. In this study, a method based on ultra-high-performance liquid chromatography coupled to high resolution mass spectrometry together with diagnostic fragment ions (DFIs) mass defect filtering (MDF) was developed to systematically screen and identify the chemical ingredients presenting in Epimedium-Astragalus granule pair (EAGP) and the absorbed components and their metabolites in rat plasma following oral administration. Using accurate mass determination, mass defect filtering and diagnostic fragment ion screening strategies, a total of eighty-five ingredients were identified in EAGP. By comparing the total ion chromatograms obtained from dosed rat plasma, blank rat plasma and EAGP solution, a total of forty-six compounds were detected in dosed rat plasma, including twenty-five prototypes and twenty-one metabolites. Among these, seventeen parent compounds were derived from Epimedium and eight were from Astragalus. These metabolites were associated with ononin (M1, M2, M9 M10 and M17), calycosin-7-<em>O</em>-<em>β</em>-D-glucoside (M6, M7, M8 and M13), icariin (M3, M4, M5, M11, M14, M15, M18, M19, M20 and M21) and methylnissolin (M12). The metabolic pathways included hydroxylation, demethylation, deglycosylation and glucuronidation. This study elucidated the potential pharmacologically active components of EAGP and provided essential data for the further study on its pharmacological materials basis and mechanism of action.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"259 ","pages":"Article 116768"},"PeriodicalIF":3.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development and validation of a UPLC-MS/MS method for almonertinib with its active metabolite HAS-719 and anlotinib in human plasma

IF 3.1 3区医学

Journal of pharmaceutical and biomedical analysis Pub Date : 2025-02-24 DOI: 10.1016/j.jpba.2025.116766

Yi Qin , Xiaojun Guan , Shichao Zhang , Zijie Zhang , Chenrong Huang , Liyan Miao

{"title":"Development and validation of a UPLC-MS/MS method for almonertinib with its active metabolite HAS-719 and anlotinib in human plasma","authors":"Yi Qin , Xiaojun Guan , Shichao Zhang , Zijie Zhang , Chenrong Huang , Liyan Miao","doi":"10.1016/j.jpba.2025.116766","DOIUrl":"10.1016/j.jpba.2025.116766","url":null,"abstract":"<div><div>Almonertinib and anlotinib are tyrosine kinase inhibitors used to treat malignant tumors, with their combination currently applied to non-small cell lung cancer (NSCLC) patients. This study aimed to develop a simple and rapid UPLC–MS/MS method for simultaneously detecting almonertinib, its active metabolite HAS-719, and anlotinib in human plasma. The analytes were separated on a Waters HSST3-C18 column following protein precipitation with acetonitrile. Mass detection was performed on a Waters TQS triple quadrupole mass spectrometer under positive electrospray ionization mode. The MRM ions were <em>m/z</em> 526.5→71.96 for almonertinib, <em>m/z</em> 512.4 → 455.41 for HAS-719, <em>m/z</em> 408.16→ 339.03 for anlotinib and <em>m/z</em> 518.5→372.26 for d6-HAS-000719 (internal standard). Method validation followed FDA guidelines and Chinese Pharmacopoeia regulations, demonstrating acceptable accuracy, precision, matrix effects, recovery, and stability. The method showed excellent linearity over 0.5–500 ng/mL for all three analytes, with correlation coefficients (r²)≥ 0.99. The validated UPLC-MS/MS method successfully monitored almonertinib and anlotinib concentrations in clinical, particularly for NSCLC patients.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"260 ","pages":"Article 116766"},"PeriodicalIF":3.1,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143549090","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0