Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Ultrasonication-assisted extraction and exploration of untargeted metabolomics in Agaricus bisporus fruiting body and stipe using LC/MS Q-TOF","authors":"Charu Agnihotri ,&nbsp;Bhim Pratap Singh ,&nbsp;Shwet Kamal ,&nbsp;Vijay Singh Sharanagat","doi":"10.1016/j.jpba.2025.117257","DOIUrl":"10.1016/j.jpba.2025.117257","url":null,"abstract":"<div><div>The present study was undertaken to compare the bioactive potential and presence of metabolites in <em>Agaricus bisporus</em> fruiting body (ABF) and <em>Agaricus bisporus</em> stipe (ABS) using Ultrasound-assisted extraction (UAE), which enhanced the extraction yield by 1.3-fold and 1.7-fold in ABF and ABS, respectively. ABF expressed a higher amount of total phenol content, which correlates with the antioxidant potential determined by DPPH, FRAP, and ABTS assays. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) determined 1225 and 1249 compounds in ABS and ABF, respectively. Among the untargeted compounds, gamma-sitosterol and phenylalanine were found abundant in ABS and ABF, respectively. Principal Component Analysis (PCA) further validated the correlation among the obtained metabolites from ABS and ABF. Furthermore, KEGG analysis was performed to understand the dominance of metaboloic pathways. To the best of our knowledge, herein we report the first comprehensive LC/MS-QTOF metabolite profiling of ABS, revealing its nutraceutical potential.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117257"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622392","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}

{"title":"Recent advances in electrochemical sensors for the detection of anticancer drugs","authors":"Sevda Hasanova ,&nbsp;Eda Gumus ,&nbsp;Erhan Zor","doi":"10.1016/j.jpba.2025.117278","DOIUrl":"10.1016/j.jpba.2025.117278","url":null,"abstract":"<div><div>The incidence and mortality rates of cancer are rapidly increasing worldwide. As the global population grows and ages, cancer has become a leading cause of death, partly due to significant reductions in mortality rates from stroke and coronary heart disease in many countries. Despite significant advances in cancer therapy, there is a substantial interest in developing new anticancer agents with different mechanisms of action, as cancer cells have developed resistance to current treatments. Noteworthy anticancer agents in this category include doxorubicin, 5-fluorouracil, and methotrexate among others. There has been a concerted effort to develop rapid, sensitive, and non-destructive methods for detecting the effects and mechanisms of anticancer drugs both in vitro and in vivo. Currently, methods such as mass spectrometry-based liquid chromatography (LC-MS), capillary electrophoresis (CE), and mass spectrometry-based gas chromatography (GC-MS) are still widely used to study anticancer drugs. However, electroanalytical techniques have recently gained popularity due to their higher sensitivity, greater selectivity, eco-friendliness, shorter analysis time, and lower cost. These techniques are now frequently used to detect anticancer drugs. Identifying anticancer drugs at low concentrations and with high sensitivity is crucial for tracking these medications. In this study, we discuss the recent advances in electrochemical sensors for the detection of anticancer drugs between 2019 and 2024.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117278"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622394","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}

{"title":"Efficient separation of chemical constituents from shilajit by an inner-recycling counter-current chromatography and their molecular docking analysis for osteoporosis","authors":"Shahid Aziz ,&nbsp;Iftikhar Ali ,&nbsp;Manzoor Hussain ,&nbsp;Sujjad Hyder ,&nbsp;Ajmal Khan ,&nbsp;Kashif Ali ,&nbsp;Salma Batool ,&nbsp;Xiao Wang","doi":"10.1016/j.jpba.2025.117274","DOIUrl":"10.1016/j.jpba.2025.117274","url":null,"abstract":"<div><div>Shilajit (<em>Asphaltum punjabianum</em>), a herbomineral ethnomedicinal substance from layers of rocks in the Himalayas and Hindukush ranges, is reported to treat urinary bladder cancer, it reduces inflammation, oxidative stress and bone loss. To date, there are no such authentic reports on the detailed separation or isolation of the chemical constituents from shilajit. In the current study, the high-speed counter-current chromatography technique is used for the separation of chemical constituents from shilajit for the first time. Thus, the current investigation depicts an efficient separation of six components namely boscartin J (<strong>1</strong>), 3<em>α</em>-<em>O</em>-acetyl-11-keto-<em>β</em>-boswellic acid (<strong>2</strong>), boscartin K (<strong>3</strong>), hemerocallal A (<strong>4</strong>), 3<em>α</em>-hydroxytirucalla-8,24-dien-21-oic acid (<strong>5</strong>), and centrolobol (<strong>6</strong>) using HSCCC employing a solvent system MTBE/n-But/ACN/H2O (2:2:1:5, v/v). Using the molecular docking, compounds <strong>2</strong> and <strong>5</strong> exhibited a significant affinity for ERα (PDB 3OCB) and Cathepsin K (PDB 6WOK). It is worth mentioning that the simulated results reflect a profound alignment with established and renowned bone-protective pharmacological principles. 3α-O-Acetyl-11-keto-β-boswellic acid and 3α-hydroxytirucalla-8,24-dien-21-oic acid exhibit promising dual activity, modulation of ERα and inhibition of Cat. K for the treatment of osteoporosis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117274"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145622389","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}

{"title":"Key considerations for phosphatidylserine quantification by 31P NMR in pharmaceutical analysis","authors":"Daniela Rebollar-Ramos,&nbsp;Shao-Nong Chen,&nbsp;David C. Lankin,&nbsp;Robert A. Kleps,&nbsp;Guido F. Pauli","doi":"10.1016/j.jpba.2025.117248","DOIUrl":"10.1016/j.jpba.2025.117248","url":null,"abstract":"<div><div>Phosphatidylserine (PS) is a phospholipid essential for cell membrane composition and with high concentrations in the brain. PS is widely used as a dietary supplement due to its neuroprotective effects and the benefits shown to patients with Central Nervous System (CNS) disorders. This work aimed to identify critical aspects in the development of a quantitative NMR (qNMR) method using quantitative ³¹P NMR (qPNMR) with triphenyl phosphate (TPP) as an internal calibrant (IC) for the analysis of PS samples. The findings indicate several pivotal aspects in the method development: i) sample preparation is a critical step for successful qNMR analysis; the addition of EDTA 0.2 M (pH 7.2–7.5, adjusted with Cs₂CO₃) enhances the lineshape and, thus, the overall resolution of the spectra; ii) the molecular weight of the analyte significantly impacts the analytical outcomes; and iii) post-acquisition processing conditions, including window functions and integration criteria, have slight but noticeable effects on the quantitative outcomes and should, therefore, be considered an integral part of any convention method for PS quantitation. To overcome this latter aspect, peak fitting was found to be an effective approach for reducing operator bias, especially in integration, and is viable due to the essential near-singlet nature of the ³¹P NMR peak patterns for each individual chemical species.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117248"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145668888","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}

{"title":"Integrating multi–wavelength HPLC fingerprinting and electrochemical profiling with chemometrics for comprehensive quality assessment of Kangfuxin Liquid","authors":"Xiang Li ,&nbsp;Ting Yang ,&nbsp;Fan Zhang ,&nbsp;Guoxiang Sun ,&nbsp;Mingxiao Hou ,&nbsp;Yunen Liu","doi":"10.1016/j.jpba.2025.117308","DOIUrl":"10.1016/j.jpba.2025.117308","url":null,"abstract":"<div><div>Kangfuxin Liquid (KFX), a complex zoogenic medicine, faces challenges in quality control due to its multi-component nature and batch-to-batch variability. This study presents a novel integrated quality assessment strategy combining Four-wavelength maximization profiling (Four-WMP) HPLC with electrochemical fingerprinting (ECFP) for comprehensive evaluation of KFX. The Four-WMP method integrated chromatographic data from four wavelengths (236, 254, 265, and 280 nm) into a unified fingerprint, significantly enhancing chemical comprehensiveness compared to conventional single-wavelength approaches. Concurrently, ECFP provided unique insights into redox-active properties. Strong correlations were established between chemical fingerprints and antioxidant efficacy through Partial Least Squares (PLS) modeling (R²=0.8672, Q²=0.789), identifying critical quality markers including protocatechuic acid and adenosine. Substantial inter-manufacturer variation was observed, particularly in sodium benzoate content, highlighting significant quality inconsistencies. This multi-parametric framework provides a holistic and robust paradigm for quality control of KFX and similar complex medicines, effectively bridging chemical composition with bioactive relevance.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117308"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733479","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}

{"title":"Characterizing the in vitro metabolic features of nine alicyclic fentanyl-type new psychoactive substances","authors":"Xuan Luo ,&nbsp;Wenting Liu ,&nbsp;Kejian Huang ,&nbsp;Xiaofeng Liu ,&nbsp;Ning Yang ,&nbsp;Qiulian Luo","doi":"10.1016/j.jpba.2025.117276","DOIUrl":"10.1016/j.jpba.2025.117276","url":null,"abstract":"<div><div>New psychoactive substances (NPS) represent a third-generation class of drugs characterized by high addictive potential, enhanced potency, low lethal doses, rapid <em>in vivo</em> metabolic rates, and fast structural diversification. Among these, fentanyl-type NPS (F-NPS), which are derived from the opioid analgesic “fentanyl”, present a category of NPS that simultaneously exhibit all five characteristics (in addition to nitazenes). Consequently, global F-NPS use poses a critical challenge, as metabolic data from F-NPS have often proven insufficient to support law enforcement agencies in combating emerging analogs. In this study, an <em>in vitro</em> human liver microsome model was used to incubate nine alicyclic F-NPS derivatives and the metabolites was analyzed by using liquid chromatography-ion trap tandem time-of-flight mass spectrometry. We systematically characterized the metabolic profiles of the alicyclic F-NPS and elucidated the structural determinants that influence metabolism. Our findings revealed that <em>sp</em>³-carbon monohydroxylation occurring at the alicyclic ring, a distinctive metabolic pathway in this subclass, is governed by the C–H Laplacian bond order, intermediate radical stability, alicyclic ring rigidity, and steric hindrance, while also competing with analogous metabolic pathways at other sites. Notably, alicyclic dehydrogenation, a previously unreported metabolic pathway, correlated with the stability of alicyclic alkenes and conjugative stabilization through adjacent carbonyl groups. <em>N</em>-oxidation metabolism was significantly modulated by <em>N</em>-dealkylation metabolism at adjacent positions and substituents on the phenyl group of the core. Furthermore, &gt;N–CH₂–CH₂– serves as the primary metabolic hotspot within the F-NPS skeleton structure, with the total relative contents of corresponding metabolites accounting for &gt; 80 % (and up to 98 %). The monohydroxylation of the phenyl group adhered to an electrophilic substitution mechanism governed by the electron density and both the theoretical calculation results and mass spectrometric data consistently indicated that <em>sp</em>²-carbon monohydroxylation occurred exclusively at the phenethyl group in the tail. The findings provide data and analytical strategies for the development of a universal F-NPS metabolic prediction model.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117276"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145661123","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}

{"title":"Chemical stability of esomeprazole in biorelevant bicarbonate buffer and an exploratory study for specific degradation products","authors":"Ayaka Takasusuki ,&nbsp;Takahiro Takayama ,&nbsp;Koichi Inoue ,&nbsp;Kiyohiko Sugano","doi":"10.1016/j.jpba.2025.117305","DOIUrl":"10.1016/j.jpba.2025.117305","url":null,"abstract":"<div><div>This study aimed to evaluate the impact of biorelevant bicarbonate buffer solutions (BCB) on drug degradation relative to phosphate buffer solutions (PPB). Esomeprazole (ESM) was selected as a model compound. Stability studies were conducted at 37 °C using 10 mM BCB and 31 mM PPB (both at pH 5.5; buffer capacity (β) = 4.0 mM/pH; ionic strength (<em>I</em>) = 0.14 M). Degradation products were quantified at 4 time points by HPLC. No significant difference was observed in the degradation rate constant of ESM between BCB and PPB. However, LC–MS analysis revealed a distinct tendency that two degradation products with <em>m/z</em> values of 330 and 298 were more abundant in BCB than in PPB. The putative structures of these degradation products matched the previously reported compounds. These findings suggest that buffer species can influence degradation pathways even under equivalent β and <em>I</em>. Consequently, the use of biorelevant buffer systems is essential for accurate assessments of degradation products.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117305"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145724126","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}

{"title":"Development of a reliable and convenient lateral flow immunoassay for dihydromyricetin detection in Ampelopsis grossedentata samples","authors":"Xueyan Cui ,&nbsp;Zhen Cao ,&nbsp;Hailong Yu ,&nbsp;Hui Li ,&nbsp;Ying Ying ,&nbsp;Ting Luo ,&nbsp;Jing Wang","doi":"10.1016/j.jpba.2025.117299","DOIUrl":"10.1016/j.jpba.2025.117299","url":null,"abstract":"<div><div>Dihydromyricetin, a flavonoid abundant in <em>Ampelopsis grossedentata</em> distributed across southern China, exhibits diverse pharmacological activities. In this study, a precise, sensitive, rapid, and reliable lateral flow immunoassay (LFIA) was developed for dihydromyricetin quantification in <em>Ampelopsis grossedentata</em>. A dihydromyricetin hapten was synthesized by introducing a carboxylic acid spacer via a four-carbon chain at the 3-position. A hybridoma cell line (2H3) secreting a highly specific monoclonal antibody (mAb) against dihydromyricetin was established. Dihydromyricetin demonstrated excellent stability in most organic solvents and acidic conditions. The LFIA exhibited a sensitivity (IC₅₀) of 33.61 ng/mL and a linear range of 4.54–281.83 ng/mL. The recovery rates of dihydromyricetin were in the range of 70.83–96.17 %. These findings indicate that LFIA, leveraging the high specificity of the dihydromyricetin mAb, offers a simple, rapid, and accurate method for dihydromyricetin analysis in <em>Ampelopsis grossedentata</em>.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117299"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145714500","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}

{"title":"Identification and extraction of bioactive constituents from Gardenia jasminoides Ellis: Evaluation of their acetylcholinesterase inhibitory potential","authors":"Xuanlin Liu ,&nbsp;Yang Zhou ,&nbsp;Ming Chen ,&nbsp;Yuchi Zhang ,&nbsp;Sainan Li ,&nbsp;Long Fei","doi":"10.1016/j.jpba.2025.117240","DOIUrl":"10.1016/j.jpba.2025.117240","url":null,"abstract":"&lt;div&gt;&lt;div&gt;Research in functional foods faces two major challenges due to the complexity of natural food components: accurate and rapid identification of bioactive compounds, and their precise extraction. To address these challenges, this study developed an integrated strategy to identify and isolate acetylcholinesterase inhibitors (AChEIs) from Gardenia jasminoides Ellis (&lt;em&gt;GJE&lt;/em&gt;)—a plant with both medicinal and edible value—while evaluating their potential against Alzheimer’s disease (AD). First, affinity ultrafiltration-liquid chromatography-mass spectrometry (AUF-LC-MS) (a technique that uses AChE as a target to specifically capture binding compounds from complex extracts) was employed to screen candidate AChEIs from &lt;em&gt;GJE&lt;/em&gt;. To verify the anti-dementia potential of these candidates and analyze their binding stability with AChE, we used molecular docking integrated with molecular dynamics simulation (a type of kinetic simulation approach that predicts static binding affinity and dynamic complex stability, respectively). Next, based on activity screening results, offline two-dimensional countercurrent chromatography (Offline 2D-CCC) (a separation technique that first groups compounds with similar polarity via 1st-dimensional separation and then purifies each group with optimized solvent systems via 2nd-dimensional separation) was used to isolate the active components. To clarify the inhibitory mechanism of the isolated compounds on AChE, we performed targeted enzymatic kinetics analysis (including determining inhibition type, calculating the half-inhibitory concentration IC₅₀, and measuring the inhibition constant K&lt;em&gt;ᵢ&lt;/em&gt;—parameters that characterize how compounds block AChE activity). To systematically confirm the anti-AD mechanism of &lt;em&gt;GJE&lt;/em&gt; components and assess potential toxic effects, we applied network pharmacology—a method that constructs “compound-target-disease” interaction networks to reveal multi-component, multi-target, and multi-pathway relationships (instead of focusing on single targets). This approach helped identify cross-talk between &lt;em&gt;GJE&lt;/em&gt;’s therapeutic effects and potential toxicity. Nine potential AChEIs were finally screened from &lt;em&gt;GJE&lt;/em&gt;: Geniposidic acid, Genipin-1-&lt;em&gt;O&lt;/em&gt;-gentiobioside, Gardenoside, Croceic acid, Rutin, Crocin I, Crocin II, Quercetin, and Nevadensin. Their binding energies with AChE (a key parameter for evaluating binding strength) were −8.8, −8.9, −8.2, −8.7, −9.3, −10.4, −10.6, −10.2, and −10.3 kcal/mol, respectively. The terpenoids and flavonoids isolated via Offline 2D-CCC exhibited &gt; 90 % purity (a critical standard for natural product research). Enzymatic kinetics analysis confirmed that these compounds exert reversible AChE inhibition—a property essential for developing safe therapeutic agents for cholinergic dysfunction disorders like AD. To improve the efficiency and accuracy of active compound screening, we combined computer-aided virtual screening (in this s","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117240"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145555397","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}

{"title":"Comprehensive characterization of the pharmacokinetics, tissue distribution, and metabolism of Bufei Formula in rats","authors":"Hou-Xu Ma ,&nbsp;Na-Wei Qi ,&nbsp;Xiao-Lin Yang ,&nbsp;Fei Li","doi":"10.1016/j.jpba.2025.117315","DOIUrl":"10.1016/j.jpba.2025.117315","url":null,"abstract":"<div><div>Bufei Formula (BF), consisting of nine traditional Chinese medicinal herbs, is clinically applied to tonify qi, nourish the lung, relieve cough, and alleviate asthma. However, the chemical complexity of BF has hindered a clear understanding of its pharmacodynamic material basis and underlying <em>in vivo</em> mechanisms. This study aims to systematically elucidate the pharmacokinetics, tissue distribution, and metabolic profiles of BF in rats, thereby laying the groundwork for understanding its pharmacodynamic material basis. First, ultra-performance liquid chromatography (UPLC) was used to identify 18 major active constituents of BF. Subsequently, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed to identify prototype compounds and metabolites in rat plasma, urine, feces, and tissues following oral administration of BF. In addition, a validated UPLC-MS/MS method was established for quantifying 14 active constituents. In total, 63 constituents were identified <em>in vivo</em>, including 17 prototype compounds and 46 metabolites. Among them, eight compounds showed the mean time to maximum plasma concentration (T_max) of 0.112–0.987 h, maximum plasma concentrations (C_max) of 4.995–213.333 μg/L, and the area under the plasma concentration–time curve (AUC) of 7.557–692.082 μg·h/L. Tissue distribution analysis revealed that the 14 quantified compounds were widely distributed in multiple organs, with predominant accumulation in the lung and colon, suggesting them as potential target organs. These findings provide a valuable reference for the clinical application of BF and evidence for elucidating its pharmacodynamic material basis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"270 ","pages":"Article 117315"},"PeriodicalIF":3.1,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733397","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}