Journal of pharmaceutical and biomedical analysis最新文献

{"title":"Pharmacokinetic investigation on eight primary active constituents of Weifuchun capsules in healthy and chronic atrophic gastritis model rats","authors":"Wentao Shao ,&nbsp;Xinxin Zhao ,&nbsp;Qiuya Zhou ,&nbsp;Lvli Ma ,&nbsp;Wei Zhang ,&nbsp;Yi Tao","doi":"10.1016/j.jpba.2026.117335","DOIUrl":"10.1016/j.jpba.2026.117335","url":null,"abstract":"<div><div>Weifuchun capsule (WC) is clinically employed in treating chronic atrophic gastritis (CAG), yet differences in its pharmacokinetic profile between normal and chronic atrophic gastritis model rats remain insufficiently characterized. In this study, a reliable and precise ultra-high performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of eight major active constituents including <em>p</em>‑coumaric acid, ginsenoside Rg1, kaempferol, luteolin, hesperetin, apigenin, naringenin, and tangeretin in rat plasma. This method was subsequently applied to investigate the pharmacokinetics of Weifuchun capsule. A chronic atrophic gastritis model was established using a modified chemical stimulation method. Plasma samples were pretreated via protein precipitation with chloramphenicol as the internal standard and then analyzed. The intra- and inter-day precision for all eight analytes was below 6.32 %, with accuracies within ±5.19 %. Extraction recoveries ranged from 89.15 % to 112.50 %, and matrix effects were between 95.69 % and 104.83 %. All analytes demonstrated satisfactory stability under various storage conditions. The validated method was successfully applied to a comparative pharmacokinetic study. Compared with the normal group, the chronic atrophic gastritis model group exhibited significantly increased C_max and AUC_0→t values for ginsenoside Rg1, hesperetin, and naringenin (<em>p</em> &lt; 0.05), along with significantly elevated C_max for kaempferol and apigenin (<em>p</em> &lt; 0.05). Additionally, t_1/2 was significantly shortened for ginsenoside Rg1 and <em>p</em>-coumaric acid. These findings suggest enhanced absorption and accelerated elimination of certain bioactive components of Weifuchun capsules under chronic atrophic gastritis pathological conditions. The altered pharmacokinetic behavior of multiple active compounds of Weifuchun capsules in chronic atrophic gastritis model rats provides important insights into the pharmacological mechanisms of Weifuchun capsules in the treatment of chronic atrophic gastritis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117335"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917927","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":"Fluorescence spectral-chromaticity mapping for non-destructive authentication of herbal medicines","authors":"Jiawei Feng ,&nbsp;Guirong Bu ,&nbsp;Xigui Song ,&nbsp;Peiyu Wu ,&nbsp;Zhiyuan Yan ,&nbsp;Shenfei Chen ,&nbsp;Qi Yao","doi":"10.1016/j.jpba.2025.117307","DOIUrl":"10.1016/j.jpba.2025.117307","url":null,"abstract":"<div><div>Identification of the authenticity of herbal medicines is crucial to ensure their safety and efficacy, yet quantitative, accurate, and practical identification methods remain limited. This work introduces a fluorescence-based approach for distinguishing authentic and counterfeit herbal medicines by analyzing their spectral and chromaticity features under 365 nm UV excitation. A hyperspectral imaging camera was employed to collect the spectral data from three groups of herbal medicines, Ziziphi Spinosae Semen (ZSS), Aurantii Fructus (AF), Carthami Flos (CF) and their common counterfeits (a total of 326 pieces of spectral data in the three groups, 158, 80, 88 respectively). The critical difference between ZSS, AF, CF and their counterfeits was obtained by extracting the fluorescence spectrum features. The mapping relationships between the spectrum features of ZSS and the colorimetric features were established by using the Fast Formula Fitting (FFF) and Neural Network Fitting (NNF) methods. The correlation coefficients of the two fitting methods in three groups were 0.985, 0.995, 0.999 and 0.992, 0.998, 0.989 respectively. The spectral power distribution in the range of 400–780 nm (good for AF, CF and their counterfeits) and 470–780 nm (good for ZSS and its counterfeits) were identified as optimal for color-based discrimination, guiding future filter selection for portable devices. Both methods enabled pixel-level authentication with high precision, supported by a defined chromaticity visualization zone. This non-destructive, quantitative technique facilitates simple and accurate herbal medicine verification, paving the way for low-cost, portable authenticity detection systems.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117307"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145760867","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":"Rapid appearance of caffeine in plasma of rats exposed to e-cigarette smoke, analyzed using a validated LC-MS/MS method","authors":"Keisuke Muranaka ,&nbsp;Junpei Mutoh ,&nbsp;Makoto Takada ,&nbsp;Yutaro Yanagita ,&nbsp;Kanta Noguchi ,&nbsp;Ken Tachibana ,&nbsp;Naoya Kishikawa ,&nbsp;Naotaka Kuroda ,&nbsp;Mitsuhiro Wada","doi":"10.1016/j.jpba.2025.117317","DOIUrl":"10.1016/j.jpba.2025.117317","url":null,"abstract":"<div><div>Although electronic cigarettes (e-cigarettes) that enable inhalation of caffeine (Caf) are commercially available, the pharmacokinetic (PK) properties of Caf intake via e-cigarette smoking are unclear. The present study used a validated LC-MS/MS method to compare the PK parameters of Caf and its metabolites (Cafs) in rat plasma after oral (<em>p.o.</em>) administration of Caf and exposure to e-cigarette smoke. The LC-MS/MS method was validated for Caf and metabolites such as paraxanthine (PX), theophylline (TP), theobromine, 1,3,7-trimethyluric acid, 3-methylxanthine, 1,3-dimethyluric acid, and 1,7-dimethyluric acid. Quantitation of Cafs in deproteinated rat plasma was achieved. Rats were exposed to e-cigarette smoke for 1 and 3 min, and then plasma was collected until 90 min. Although the Caf exposure dose for rats was estimated as very low, quantitation of Caf, PX, and TP was possible using the proposed LC-MS/MS method. The maximum concentrations of Caf after 1 and 3 min of exposure to e-cigarette smoke were similar to those observed after <em>p.o</em>. administration of 40 and 90 μg/kg, respectively. Furthermore, PK parameters such as the concentration peak time (3.0 min for 1- and 3-min exposure) and absorption rate constant (0.62 for 1 min and 0.91 min⁻¹ for 3 min) for e-cigarette smoke exposure differed significantly from p.o. administration (50.0 min and 36.7 min, and 0.045 and 0.070 min⁻¹) (<em>P</em> &lt; 0.05). By contrast, no difference was observed between PX and TP. This is the first report demonstrating the rapid appearance of Caf in rat plasma after exposure to e-cigarette smoke.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117317"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145781430","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":"Determination of N-Lactoyl-phenylalanine, phenylacetylglutamine and trimethylamine-N-oxide related metabolites in hypertensive patients by liquid chromatography-mass spectrometry and the application to blood pressure regulation","authors":"Xin Tan ,&nbsp;Xianyun Jiang ,&nbsp;Lu Ren ,&nbsp;Ziyu Meng ,&nbsp;Jingyi Yang ,&nbsp;Yuan Wang","doi":"10.1016/j.jpba.2025.117327","DOIUrl":"10.1016/j.jpba.2025.117327","url":null,"abstract":"<div><div>Metabolites, such as N-lactoyl-phenylalanine (Lac-Phe), phenylacetylglutamine, and trimethylamine-N-oxide related precursors, may be associated with hypertension. A rapid, sensitive and accurate method, with high throughput, low sample volume, and easy operation to analyze these metabolites would facilitate their measurement and provide important insights into their role in hypertension. We developed an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous measurement of eight metabolites in 5 µL human plasma samples with a 4.2-min chromatographic run and easy operation. The method achieved good linearity (R² &gt; 0.99), sensitivity (limit of quantification: 0.1–1 ng/mL), accuracy (within-day: −5.33 % to 11.38 %; between-day: −3.35 % to 11.50 %), precision (within-day: 1.35–13.25 %; between-day: 1.74–12.73 %), matrix effect (−5.93 % to 14.81 %), recovery rates (91.49 % to 103.31 %), bench-top stability (coefficient of variations [CVs]: 1.02–6.16 %), long-term stability (CVs: 3.63–12.94 %), on-instrument stability (CVs: 0.53–5.09 %) and freeze-thaw stability (biases: −12.60 % to −0.13 %). We successfully utilized the UHPLC–MS/MS method to measure plasma samples obtained from 204 healthy individuals and 204 patients with hypertension. Plasma Lac-Phe was reduced in patients with hypertension, and the potential effects of blood pressure regulation by Lac-Phe were explored. In angiotensin II-induced hypertensive mice, Lac-Phe alleviated the increases in systolic and diastolic blood pressure. These findings highlight the therapeutic potential of Lac-Phe in hypertension.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117327"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145881327","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":"Banxia Xiexin Decoction modulates short-chain fatty acid metabolism and mitigates ulcerative colitis by reshaping the intestinal microbiota","authors":"Ji Wanli ,&nbsp;Song Xingyue ,&nbsp;Liang Xinning ,&nbsp;Zheng Yashuo ,&nbsp;Qian Xiaojing ,&nbsp;Hu Cheng ,&nbsp;Huo Yan","doi":"10.1016/j.jpba.2025.117332","DOIUrl":"10.1016/j.jpba.2025.117332","url":null,"abstract":"<div><div>Ulcerative colitis (UC) is a chronic inflammatory disorder that significantly impacts the quality of life for patients. Dysbiosis of gut microbiota and changes in short-chain fatty acid (SCFA) metabolism play a role in both the initiation and progression of UC. Banxia Xiexin Decoction (BXD), a formula in traditional Chinese medicine, has shown therapeutic effects; however, its underlying mechanism remains unclear. A rat model of colitis induced by dextran sulfate sodium (DSS) was created, and various doses of BXD were administered. The principal components of BXD were analyzed through high-pressure liquid chromatography (HPLC). In order to clarify the mechanisms, 16S rRNA sequencing, serum metabolomics, targeted profiling of SCFAs via gas chromatography-mass spectrometry (GC-MS), and assessments of enzyme activity were conducted. BXD exerted protective effects against DSS-induced UC, as indicated by attenuated histological damage and reduced expression of pro-inflammatory cytokines. Untargeted metabolomics revealed that BXD modulated multiple metabolic pathways, enhancing SCFA-related processes, such as propanoate and butanoate metabolism. GC–MS revealed that BXD could increase the level of acetate, propionate, butyrate, isobutyrate, valerate, isovalerate and hexanoate. Microbiome sequencing indicated that BXD increased the abundance of beneficial taxa (e.g. Firmicutes, Bacteroidetes), while reducing pro-inflammatory genera. Altogether, BXD restored the microbial balance and promoted anti-inflammatory metabolite production. Our results demonstrated that BXD ameliorated UC by reprogramming gut microbial composition and enhancing SCFA biosynthesis, thereby suppressing intestinal inflammation. The potential of BXD as a therapy aimed at the microbiota for UC is emphasized in these studies, which also offer mechanistic insights using multi-omics approaches.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117332"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917938","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":"A standard-free strategy for the differentiation of isomers and deep profiling of chemicalome in Chinese medicinal formulas","authors":"Ying-Shan Li ,&nbsp;Xing-Ting Dong ,&nbsp;Gui-Zhong Xin ,&nbsp;Li-Fang Liu ,&nbsp;Zi-Qi Shi","doi":"10.1016/j.jpba.2026.117349","DOIUrl":"10.1016/j.jpba.2026.117349","url":null,"abstract":"<div><div>The precise differentiation of isomers is crucial for comprehensive mass spectrometry-based analysis of complex Chinese medicinal formulas (CMFs). To address this challenge, a standard-free strategy was proposed by integrating chromatographic behavior, mass spectrometric behavior, and quantum chemical calculations. Firstly, structural isomers and certain stereoisomers were differentiated based on characteristic fragment ions and calculated partition coefficient (<em>clogP</em>). For remaining indistinguishable isomers, fragment ions with significant intensity differences were identified, and 3D molecular structures of the precursor ions, product ions, and neutral loss fragments were constructed. The bond dissociation enthalpies (BDE) for critical fragmentation pathways were computed using density functional theory (DFT) and correlated with relative fragment ion abundances to achieve differentiation, following the principle that higher cleavage energies correspond to reduced fragmentation efficiency and lower product ion intensities. The methodology was validated using limited standards and the optimal collision energy (OCE) fitting. Using Qiangli Pipa syrup as a case, this integrated approach enabled the identification of 203 compounds, including 40 isomer groups comprising 27 structural isomers and 13 stereoisomers. This strategy overcomes the limitations of conventional methods that are typically restricted to specific structural classes, demonstrating robust capability for systematic isomer differentiation in CMFs.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117349"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977943","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":"Systemic exposure characteristics and pharmacokinetic studies of intranasally delivered qinhao nasal drops by UHPLC-Q/TOF-MS and UHPLC-MS/MS","authors":"Maojie Zhou ,&nbsp;Mireyi Bahatijiang ,&nbsp;Wenkang Liu ,&nbsp;Junyi Wang ,&nbsp;Yan Mao ,&nbsp;Zhengyi Gu ,&nbsp;Changhong Wang","doi":"10.1016/j.jpba.2025.117320","DOIUrl":"10.1016/j.jpba.2025.117320","url":null,"abstract":"<div><div>Qinhao nasal drops (QHND) is a formulation developed based on the integration theory of ethnomedical theory and traditional Chinese medicine, which plays a significant role in the treatment of acute, chronic, and allergic rhinitis. However, its primary active constituents and <em>in vivo</em> pharmacokinetic profile remain unclear. This study aimed to characterize the chemical composition of QHND using ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry, as well as to identify the systemic circulating components following intranasal administration of QHND. The results showed that 78 chemical constituents were identified in QHND, including flavonoids, terpenoids, alkaloids, and others. Notably, 19 prototype compounds and their 12 phase I/II metabolites were detected in rat plasma. Further, a sensitive, robust, and accurate ultra-high-performance liquid chromatography-tandem mass spectrometry method was established for the quantitative determination of rupestonic acid (RA), casticin (CT), baicalin (BC), wogonin (WG), ephedrine (EP), and pseudoephedrine (PE) in rat plasma. The method underwent comprehensive validation (including selectivity, linearity, precision, accuracy, matrix effect, extraction recovery, stability, and dilution integrity), confirming its suitability for the intended applications. With this validated method, the plasma concentration-time profiles of eight target components were monitored following both intranasal and intragastric administration of QHND. The results revealed that intranasal delivery significantly reduced the onset time of pharmacological effects, facilitated more efficient and rapid systemic absorption, and subsequently enhanced the bioavailability of key active constituents (RA, EP, and PE) in rats.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117320"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145838482","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":"Revolutionizing ELISA development: The transformative impact of automation on monoclonal antibody potency assays","authors":"Xiaoran Xin ,&nbsp;Douglas Pung ,&nbsp;Amy Hsu ,&nbsp;Monisha Dey ,&nbsp;Qian Wu ,&nbsp;Bo Feng ,&nbsp;Dengyun Sun ,&nbsp;Kevin Gurney ,&nbsp;Anka Ehrhardt","doi":"10.1016/j.jpba.2025.117319","DOIUrl":"10.1016/j.jpba.2025.117319","url":null,"abstract":"<div><div>Enzyme-linked immunosorbent assays (ELISA) are a fundamental technique for determining the potency of monoclonal antibodies (mAbs), yet the traditional ELISA development process is often time-consuming and labor-intensive. In this study, we explored how automation and miniaturization in a 384-well format can revolutionize ELISA development, significantly enhancing throughput and accuracy. By employing automated liquid dispensing technology, we streamlined the optimization of assay parameters, leading to faster and more reliable potency measurements. Our findings not only demonstrate the feasibility of this innovative approach but also highlight its potential to transform the landscape of analytical assays in biopharmaceutical development.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117319"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145978445","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 analytical set-up for the monitoring of albumin adduction on Cysteine 34 exposed to mustard agents with optimized digestion and on-line SPE-LC-MS analysis","authors":"Lorenzo Avigo ,&nbsp;Marine Charpin ,&nbsp;Audrey Combès ,&nbsp;Charlotte Desoubries ,&nbsp;Christine Albaret ,&nbsp;Anne Bossée ,&nbsp;Emmanuel Joubert ,&nbsp;Valérie Pichon","doi":"10.1016/j.jpba.2025.117324","DOIUrl":"10.1016/j.jpba.2025.117324","url":null,"abstract":"<div><div>Adduction of mustard agents such as sulfur mustard (SM), sesquimustard (Q), and nitrogen mustards (HN-1, HN-2, HN-3) on nucleophilic sites of proteins, particularly Cysteine34 of human serum albumin (HSA), can be monitored in plasma to confirm retrospective exposure. The main purpose of this work is to optimize an <em>on-line</em> solid phase extraction (SPE) of HSA tripeptides containing adducted Cys34, coupled to LC-MS. To improve sensitivity of these biomarkers’ detection and data robustness the impact of sample preparation steps such as protein precipitation and reduction with dithiothreitol (DTT) prior to digestion with proteinase K (Prot. K) were systematically evaluated. Neither improved digestion efficiency, and at low incubation levels of SM, precipitation even reduced signal intensity. Removing these steps decreased variability and consumable use. Washing conditions were optimized to favour the retention of adducted tripeptides on the SPE sorbent while removing more polar digest compounds. Optimal extraction conditions defined using spiked digests involved washing with 7 % (v/v) acetonitrile (ACN) before direct on-line transfer to LC-MS. The optimized method was then applied, as a proof of concept, to plasma incubated with SM and HN-1 at two concentration levels in order to demonstrate its applicability to real biological samples. Limits of quantification (LOQs), determined within this proof-of-concept framework, were established at 10 ng/mL for SM and 40 ng/mL for HN-1 in <em>in vitro</em> adducted plasma.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117324"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145838483","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":"Separation of mephentermine from other structural analogues of phenethylamine and amphetamine in biological matrices using Liquid Chromatography-Tandem Mass Spectrometry","authors":"Tusha Tripathi ,&nbsp;Kamna Sharma ,&nbsp;Awanish Upadhyay ,&nbsp;Ashok Singh ,&nbsp;Puran Lal Sahu ,&nbsp;Tarun Handa ,&nbsp;Meghna Choudhary ,&nbsp;Bikash Medhi","doi":"10.1016/j.jpba.2025.117328","DOIUrl":"10.1016/j.jpba.2025.117328","url":null,"abstract":"<div><div>Stimulants exert euphorigenic and relaxing effects which warrant intervention of regulatory authorities in non-medical usage of these doping agents. Thus, a semi-quantitative method utilizing Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS) was developed to monitor potential misuse of 10 structural analogues of amphetamine and phenylethylamine derivatives. The extraction protocol for urine was dilute and shoot (DS) with 1 mL aliquot size, while that for plasma/serum was solid-phase extraction (SPE) with a 100 µL aliquot size. Chromatographic separation was achieved over an 18-minute gradient on a Phenomenex Luna Omega UHPLC C18 column. The mobile phase consisted of water and acetonitrile, both containing formic acid as a modifier, specifically at 1 % (v/v) for urine and 0.1 % (v/v) for plasma/serum analyses. Positive electrospray ionization (ESI+) was employed for mass spectrometric detection with acquisition in multiple reaction monitoring (MRM) mode. Method validation was performed in accordance with World Anti-Doping Agency (WADA) requirements for analyzing prohibited substances with minimum reporting limits (MRLs). Limits of identification (LOI) determined for urine and plasma/serum matrices were as follows: 0.5 ng/mL and 1 ng/mL for Oxethazaine; 1 ng/mL and 2.5 ng/mL for BHPT and BHMPT; 12.5 ng/mL and 25 ng/mL for PT and 5 ng/mL and 12.5 ng/mL for MPT, NEAMP, DMAMP, 4MMAMP, NEBMPEA and NNBTPEA, respectively. An analytical method characterized by rapidity, simplicity and cost-effectiveness was implemented for the chromatographic separation of stimulant structural analogues, facilitating the determination of potential stimulant abuse within human performance sports.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"271 ","pages":"Article 117328"},"PeriodicalIF":3.1,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145926823","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}