Journal of Chromatography B最新文献

{"title":"Preparation of the immobilized α1A-adrenoceptor column by the ultra-high affinity protein pair CL7/Im7 and its application in drug-protein interaction analysis","authors":"Qiuyu Gao ,&nbsp;Shuangru Wan ,&nbsp;Xinchao Cao ,&nbsp;Yao Chen ,&nbsp;Ning Wang ,&nbsp;Xia Wang ,&nbsp;Yue Ma ,&nbsp;Di Zhang ,&nbsp;Jing Wang ,&nbsp;Dalong Zhi","doi":"10.1016/j.jchromb.2025.124478","DOIUrl":"10.1016/j.jchromb.2025.124478","url":null,"abstract":"<div><div>Immobilizing the target protein on a solid surface with controlled orientation, high specificity, and maintained activity is a proven strategy to enhance the stability of the protein. In this study, we employed an ultra-high affinity protein pair consisting of a mutant of colicin E7 Dnase and its corresponding inhibitor, immunity protein 7(Im7), to develop an immobilized α_1A-adrenoceptor (α_1A-AR) column. Briefly, we expressed α_1A-AR fused with CL7 as a tag at its C-terminus in <em>Escherichia coli</em> cells. Meanwhile, we got His-tagged Im7 at the same manner. Following purification, the His-tagged Im7 was utilized to functionalize the macro-porous silica gel. Leveraging the ultra-high affinity between the protein pair, we achieved robust and specific covalent immobilization of α_1A-AR covalently at ambient conditions in buffer solutions, without the requirement for additional regents. The successful immobilization of the receptor, without extraneous protein adsorption, was confirmed through X ray photoelectron spectroscopy and chromatographic investigations. Frontal analysis and adsorption energy distribution analysis further validated the feasibility of the immobilization method. Our findings align well with those reported in the literature. This work is poised to provide a modular platform for conducting effective investigations into the binding interactions between other functional proteins and the drugs.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124478"},"PeriodicalIF":2.8,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143063041","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":"Potential for application of direct thrombin inhibitors isolated from Euphorbia resinifera O.Berg latex in fibrin clot formation","authors":"Jaruwan Siritapetawee ,&nbsp;Yanling Hua ,&nbsp;Chutima Talabnin ,&nbsp;Nopporn Naewwan ,&nbsp;Ratana Charoenwattanasatien ,&nbsp;Chalermluck Phoovasawat ,&nbsp;Supawan Srichan ,&nbsp;Chortip Kantachot","doi":"10.1016/j.jchromb.2025.124480","DOIUrl":"10.1016/j.jchromb.2025.124480","url":null,"abstract":"<div><div>Direct thrombin inhibitors (designated as EuRL-DTIs) were partially purified from ethanol extracts of <em>Euphorbia resinifera</em> O.Berg latex. The obtained EuRL-DTIs comprised four major compounds: two isomers of phenolic compounds (C₁₉H₂₆O₁₂) and two amide compounds (tentatively identified as C₂₄H₄₄N₄O₄ and C₃₆H₆₆N₆O₆), as identified by liquid chromatography and electrospray ionisation quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS), attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy, and/or nuclear magnetic resonance (NMR) spectroscopy. The effects of EuRL-DTIs on human thrombin-induced fibrin clot production were analysed using thrombin time, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), synchrotron radiation X-ray tomographic microscopy (SRXTM), and scanning electron microscopy (SEM). Kinetic studies revealed that EuRL-DTIs inhibited human thrombin from cleaving the chromogenic substrate S2238, with a <em>K</em>_i of 3.7 μg/mL, in a non-competitive inhibition manner. All results supported the hypothesis that the EuRL-DTIs directly abolished thrombin activity in the production of fibrin clots without requiring a cofactor. The cytotoxicity test showed that EuRL-DTIs were nontoxic to normal human foetal lung fibroblasts (IMR-90). Thus, EuRL-DTIs have potential as antithrombotic agents for application as drugs for thrombosis treatments or in medical devices such as coating surgical sutures.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124480"},"PeriodicalIF":2.8,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143063039","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 and validation of a high-performance liquid chromatography method with fluorescence detection for the quantification of the resistance protein P-gp in cancer cells","authors":"Elodie Gay,&nbsp;Maxime Dubois,&nbsp;Manon Roux,&nbsp;Antoine Goisnard,&nbsp;Marie Depresle,&nbsp;Mahchid Bamdad,&nbsp;Pierre Daumar,&nbsp;Emmanuelle Mounetou","doi":"10.1016/j.jchromb.2025.124475","DOIUrl":"10.1016/j.jchromb.2025.124475","url":null,"abstract":"<div><div>A method using high-performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) was developed and validated to quantify the innovative tool LightSpot®-FL-1, a selective permeability-glycoprotein (P-gp)-targeted fluorescent conjugate used to measure P-gp expression in cell samples. Quantifying P-gp is a major challenge in oncology as its overexpression in many cancer cells results in Multidrug Resistance (MDR) associated with chemotherapy failure. To develop the method reported herein, both sample preparation and analysis parameters were investigated. Optimal chromatographic conditions were achieved with 5 µL injections at a 1 mL/min flow rate on a reversed-phase Zorbax® Eclipse Plus 3.5 µm C18 column (150 × 4.6 mm) with isocratic acetonitrile/water (85/15, by volume) elution. Detection was performed with 505 nm excitation and 510 nm emission wavelengths. Validation studies were designed and performed according to the International Council for Harmonization (ICH) guidelines for bioanalytical method validation. The limit of quantification (LOQ) and limit of detection (LOD) were determined to be 0.5 and 0.2 nmol/L, respectively. The linearity range was demonstrated between 10 and 500 nmol/L, and the trueness and precision of the method were validated. Good stability was shown in three relevant analytical conditions. The greenness of the developed method was also demonstrated with the AGREE, AGREEprep and MoGAPI tools. Finally, the rapid, precise and sensitive validated analytical method was successfully applied to determine the difference in P-gp expression in three cancer cell lines: DU4475, CCRF-CEM and KG-1a.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124475"},"PeriodicalIF":2.8,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035739","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}

{"title":"Analysis of glipizide and pravastatin sodium drug molecules at trace level by using magnetic solid phase extraction and HPLC-DAD system","authors":"Ümmügülsüm Polat ,&nbsp;Halil İbrahim Ulusoy ,&nbsp;İsmail Murat Palabıyık","doi":"10.1016/j.jchromb.2025.124477","DOIUrl":"10.1016/j.jchromb.2025.124477","url":null,"abstract":"<div><div>An easy applicable and selective sample preparation technique has been developed for trace and simultaneously analysis of Glipizide (GLP) and Pravastatin (PST) molecules in biological matrices based on magnetic solid phase extraction (MSPE) and high-performance liquid chromatography (HPLC). A new magnetic adsorbent including Fe₃O₄@TEOS-Melamine has been synthetized and characterized for extraction studies. Experimental variables of MSPE were examined and optimized step by step such as pH, adsorption and desorption conditions, time effect, etc. Simultaneous HPLC analysis of GLP and PST molecules was performed by isocratic elution of a mixture of acetonitrile: phosphate buffer (pH:3.0, 0.02 M) (35:65) with flow rate 1.0 mL min⁻¹. Analytical signals obtained from DAD detector were recorder in 238 and 226 nm for PST and GLP, respectively. The limit of detections (LOD) for proposed method were 3.17 ng mL⁻¹ for PST and 3.03 ng mL⁻¹ for GLP molecules. Consequently, accuracy and precision of developed method were tested by means of recovery tests in synthetic urine and saliva samples. RSD % values were lower than 6.1 % and recovery values were in the range of 98.5–103.3 % for both molecules.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124477"},"PeriodicalIF":2.8,"publicationDate":"2025-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143051215","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 novel HPLC-HRMS method for quantitative analysis of resorcinol in urine: Application to hairdressers’ occupational exposure","authors":"Amandine Cambrai-Erb ,&nbsp;Flavien Denis ,&nbsp;Romain Pons ,&nbsp;Anca Radauceanu ,&nbsp;Sophie Ndaw ,&nbsp;Nathalie Grova","doi":"10.1016/j.jchromb.2025.124472","DOIUrl":"10.1016/j.jchromb.2025.124472","url":null,"abstract":"<div><div>Resorcinol is a widespread substance used in a large variety of manufacturing industries, including cosmetics, with endocrine-disrupting activity on the thyroid function. The aim of the present study was to develop and validate a sensitive, selective and robust method to quantify resorcinol in urine and thereby assess hairdressers’ occupational exposure. As resorcinol is mainly excreted in urine as glucuronide or sulfate forms, the first step consisted in hydrolyzing urine samples with a β-glucuronidase-arylsulfatase enzyme for 16 h. Then, after cleaning with a supported-liquid extraction cartridge, the samples were derivatized with dansyl chloride to improve signal and signal-to-noise ratio. Analysis was carried out using an accurate high-resolution liquid chromatography-mass spectrometry instrument on a Kinetex Biphenyl analytical column. Particular attention was paid to the chromatographic separation of resorcinol from its two isomers, catechol and hydroquinone, also present in urine. Acquisition was performed in positive ESI mode, at <em>m</em>/<em>z</em> 577.14615 for dansylated resorcinol and <em>m</em>/<em>z</em> 581.17126 for dansylated resorcinol-d₄, with respective retention times of 8.63 and 8.60 min. The method passed all the performance tests included in the validation process. The lowest limit of quantification (LLOQ) was 0.3 µg/L resorcinol, which was sufficient to quantify resorcinol in all samples tested. The calibration curves were linear from LLOQ to 2000 µg/L, with coefficients of determination R² ranging from 99.82 % to 100 %. The method was accurate, reaching 95.6 to 101.7 % of target intraday and 99.8 to 105.0 % interday, and precise with RSDs between 0.88 and 1.99 % intraday and with RSDs between 1.75 and 8.65 % in interday assessments. It also proved robust, with a matrix effect of 8.25 %. Resorcinol stability was determined by studying long-term stability at −20 °C for sample storage up to 6 months, short-term stability (at + 20 °C and + 4°C for possible short-term storage), freeze–thaw cycles, and post derivatization stability. This method was successfully applied on samples from 17 women working as hairdressers. Urinary resorcinol concentrations ranged from 2 µg/L to 1824 µg/L (6 to 4475 µg/g creatinine) and were in line with those reported in the literature.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124472"},"PeriodicalIF":2.8,"publicationDate":"2025-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035742","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}

{"title":"Preparation of halloysite nanotube-based monolithic column for small molecules and protein analysis","authors":"Qian Zhao ,&nbsp;Huixuan Li ,&nbsp;Yuanyuan Guo ,&nbsp;Moqiong Duan ,&nbsp;Mu Li ,&nbsp;Tao Li ,&nbsp;Hongya Li ,&nbsp;Shuna Li ,&nbsp;Shuxiang Wang ,&nbsp;Quan Wang","doi":"10.1016/j.jchromb.2025.124476","DOIUrl":"10.1016/j.jchromb.2025.124476","url":null,"abstract":"<div><div>s: This study aimed to prepare a new separation medium, silane coupling agent KH570- modified halloysite nanotube (MPS-HNT) monolithic column, with excellent separation performance for small molecular compounds and macromolecular proteins. This was prepared using the principle of redox polymerization with modified HNTs as monomers. The optimal monomer proportion was obtained by optimizing the ratio of monomer, cross-linker, and pore-forming agent, which was evaluated using scanning electron microscopy, nitrogen adsorption, and mercury intrusion. The monolithic column exhibited a relatively homogeneous pore structure and good separation performance and permeability. As a high-performance liquid chromatography stationary phase, six small aromatic molecules were successfully separated in 6 min with a theoretical plate count of 35, 640 plates m⁻¹ for benzene. Twenty-one peaks were separated from the fermentation mattress extract containing lipopeptide antibiotics on the MPS-HNT column. The separated chromatographic peaks further identified three lipopeptide antibiotics using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. Twelve chromatographic peaks were isolated from chicken egg whites, and the eighth peak was identified as the lysozyme. The methodological validation of the lysozyme in chicken egg white showed that the linear correlation coefficient was 0.9996. The intraday and inter-day relative standard deviations were 3.1–4.0 % and 1.9–3.4 %, respectively. The spike recovery rate of lysozyme was 94.20–103.31 %. The overall column displayed good stability: the relative standard deviations of the peak area of six aromatic compounds and retention time were &lt; 3.28 %.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1253 ","pages":"Article 124476"},"PeriodicalIF":2.8,"publicationDate":"2025-01-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143035438","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":"Screening of ESR2-targeted anti-postmenopausal osteoporosis chemistry from Rehmanniae Radix Preparata based on affinity ultrafiltration with UPLC-QE-Orbitrap-MS","authors":"Shuo Wang ,&nbsp;Yawen Li ,&nbsp;Nanxi Zhang ,&nbsp;Peitong Wu ,&nbsp;Xueqin Feng ,&nbsp;Xiaochen Gao ,&nbsp;Jiaming Shen ,&nbsp;Wanjie Liu ,&nbsp;Wei Feng ,&nbsp;Jiaming Sun","doi":"10.1016/j.jchromb.2024.124419","DOIUrl":"10.1016/j.jchromb.2024.124419","url":null,"abstract":"<div><div><em>Rehmanniae Radix Preparata</em>, a processed form of the traditional Chinese medicinal plant <em>Rehmannia glutinosa Libosch</em><u>,</u> has long been valued for its medicinal properties and use as a food. It is notably effective in treating postmenopausal osteoporosis. This study utilized C18 to separate and purify different concentrations of its eluent streams. MC3T3-E1 cells were utilized to identify the optimal ESR2 activity fraction from various concentrations of <em>Rehmanniae Radix Preparata,</em> using osteoprotegerin (OPG) as an indicator. A single-target affinity ultrafiltration method was created, combining ESR2 affinity ultrafiltration with liquid chromatography-mass spectrometry (LC-MS). Molecular docking validated the interaction mechanism between small molecule ligands and ESR2 protein. These ligands were then tested in MC3T3-E1 cells to assess survival rate, OPG content, and alkaline phosphatase (ALP) activity, an osteogenic differentiation marker. The study showed that <em>Radix Rehmanniae Praeparata</em> effectively combats PMOP, and the combined method of single-target-affinity ultrafiltration-LC-MS with molecular docking offers a robust approach for identifying its anti-PMOP compounds.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"Article 124419"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142845420","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}

{"title":"Dispersive solid phase extraction of apixaban from human plasma samples prior to capillary electrophoresis determination using zirconium-based metal organic frameworks prepared by different modulator and solvent","authors":"Aysa Abbasalizadeh ,&nbsp;Mohammad Reza Afshar Mogaddam ,&nbsp;Mir Ali Farajzadeh ,&nbsp;Mahboob Nemati ,&nbsp;Saeed Mohammad Sorouraddin","doi":"10.1016/j.jchromb.2024.124417","DOIUrl":"10.1016/j.jchromb.2024.124417","url":null,"abstract":"<div><div>Here, a zirconium-based metal organic framework–dispersive solid phase extraction method was established as an efficient, robust, and accurate approach for quantifying apixaban<!--> <!-->in human plasma samples prior to capillary electrophoresis with diode array detection. Various types of metal organic frameworks based on UiO-66-NH₂ were synthesized by altering modulators and solvents and applied as sorbents in the extraction procedure. Among the tested sorbents, UiO-66-NH₂ prepared in dimethylformamide in the presence of acetic acid was found to be the best sorbent in this method for the extraction of apixaban with high extraction efficiency comparable to other types of UiO-66-NH₂ metal organic frameworks. The extraction and preconcentration of apixaban were carried out by adding 5 mg of synthesized sorbent to a 5 mL sample solution, followed by vortexing for 3 min. After discarding the supernatant, the adsorbed analyte was eluted from the sorbent surface using 60 µL acetonitrile under vortexing for 2 min. The effective parameters of the offered method were optimized and validated using a one-parameter-at-a- time strategy. The detection and quantification limits of the method were 9.9 and 32 ng mL⁻¹ in plasma and 1.5 and 4.9 ng mL⁻¹ in deionized water, respectively. The method was linear ranging from 4.9 to 1000 ng mL⁻¹ in deionized water and from 32 to 500 ng mL⁻¹ in plasma, respectively. The enrichment factor and extraction recovery values were 44 % and 53 %, respectively. The relative standard deviations were ≤6.2 % for intra- and inter-day precisions. Finally, the proposed method was successfully employed to quantify apixaban in human plasma samples.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"Article 124417"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870647","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":"Simultaneous separation and detection of common chiral and achiral metabolites in the urine of human exposed to benzene series by LC-MS/MS","authors":"Liang Li ,&nbsp;Gang Li ,&nbsp;Huiying Xie ,&nbsp;Zhi Zhang","doi":"10.1016/j.jchromb.2024.124428","DOIUrl":"10.1016/j.jchromb.2024.124428","url":null,"abstract":"<div><div>Benzene, toluene, and xylene (BTX) are priority pollutants known for their hematotoxicity and carcinogenic properties. Benzene is further metabolized to phenyl mercapturic acid (PMA), toluene and xylene also generate benzyl mercapturic acid (BMA) in human urine. To confirm whether the exposure to benzene series comes from the workplace or from the external environment such as smoking is a very meaningful work, so accurate measurement of their biomarkers in biological samples is crucial. This study developed a novel chiral stationary phase using 6-ethylenediamine mono-derivatized-β-cyclodextrin for the simultaneous separation and detection of four chiral and achiral biomarkers PMA, BMA, N-acetyl-S-(3,4-dihydroxybutyl)-<em>L</em>-cysteine (DHBMA), and N-acetyl-S-(1-hydroxymethyl-2-propenyl)-<em>L</em>-cysteine (MHBMA) in human urine. The method demonstrated high sensitivity with detection limits below 0.211 μg/L for PMA, 0.467 μg/L for BMA, 0.246 μg/L for DHBMA, and 0.109 μg/L for MHBMA, excellent recoveries ranging from 78 to 116 % as well as the high resolutions of PMA, BMA and MHBMA-2 enantiomers being up to 1.62, 2.23 and 1.79, respectively, within 50 min under reversed-phase chromatography. Application to 60 urine samples from an automobile manufacturing plant revealed that benzene as a paint solvent has been strictly limited, while toluene and xylene are widely used, potentially posing health risks to occupational groups. The simultaneous detection of PMA, BMA, MHBMA, and DHBMA provides a more accurate assessment of non-smoking populations, enhancing the evaluation of occupational exposure to benzene series compounds.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"Article 124428"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870651","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":"Comparison of urinary excretion patterns among exposures to cosmetic preservative, herbicide, and nootropic stimulant in anti-doping analysis","authors":"I-Wen Lu ,&nbsp;Mei-Chich Hsu ,&nbsp;Yu-Tse Wu ,&nbsp;William Chih-Wei Chang","doi":"10.1016/j.jchromb.2024.124430","DOIUrl":"10.1016/j.jchromb.2024.124430","url":null,"abstract":"<div><div>Doping with meclofenoxate, a nootropic stimulant prohibited in-competition by the World Anti-Doping Agency (WADA), is identified through the primary marker of urinary 4-chlorophenoxyacetic acid (4-CPA). However, the presence of 4-CPA can also arise from permissible sources. This study ventured into comparing urinary excretion patterns among exposures to permitted chemicals (chlorphenesin and 4-CPA) and the banned stimulant (meclofenoxate) and interpreting the analytical findings according to the reporting requirements. A validated method, utilising direct injection and ultra-performance liquid chromatography–tandem mass spectrometry, was employed for urine analysis. In the first experiment, participants applied chlorphenesin-containing cosmetics with varied functions, dosages, frequencies, and application sites. Sunscreen usage led to significantly higher urinary 4-CPA concentrations (up to 1049 ng/mL) as compared to others, highlighting the impact of cosmetic formulation composition for chlorphenesin delivery. The diagnostic marker for preservative exposure included 3-(4-chlorophenoxy)-2-hydroxypropanoic acid (4-CPP) and chlorphenesin and its conjugated metabolites, with 4-CPP reaching higher concentrations (C_max of 903–7629 ng/mL) and for a longer period, up to 7–14 days. In the second experiment involving meclofenoxate supplement administration, urinary C_max levels of 4-CPA were observed between 36,287 and 39,769 ng/mL at 3–10 h post-dosing, with the parent meclofenoxate undetected in all participants’ samples. The third experiment, focused on occupational herbicide exposure in agricultural environments, detected minimal 4-CPA (&lt; 10 ng/mL) in urine. WADA’s current guidance for meclofenoxate aligns with reporting correct analytical results. Investigations, such as the experimental approach herein, offer valuable evidence addressing accuracy concerns in anti-doping tests, contributing insights for future amendments.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"Article 124430"},"PeriodicalIF":2.8,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870611","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}