Analytical and Bioanalytical Chemistry最新文献

{"title":"Intrinsic OPTIR imaging and spectroscopy enable label-free monitoring of viral infection in living cells.","authors":"Shravan Raghunathan, Kristina Worch, Christoph Krafft, Volker Deckert, Antje Burse, Juergen Popp, Tanveer Ahmed Shaik","doi":"10.1007/s00216-026-06801-x","DOIUrl":"https://doi.org/10.1007/s00216-026-06801-x","url":null,"abstract":"<p><p>Resolving biochemical signatures associated with viral infection progression within individual living cells, without introducing fluorescent labels that demand genetic accessibility and may perturb the process under study, remains a challenge for conventional optical approaches. Optical photothermal infrared (OPTIR) spectroscopy addresses this by providing label-free vibrational contrast with molecular bond specificity in live cells, combining submicron spatial resolution with strong suppression of the water background in aqueous media. Here, OPTIR discrete wavenumber imaging and point spectroscopy were applied to follow baculovirus infection in live Spodoptera frugiperda (Sf) 9 cells carrying no fluorescent reporter. Cells were measured within a CaF₂ sandwich configuration at 3, 6, 12, and 24 h post-infection. Imaging at the amide II protein band (1545 cm⁻¹) and a nucleic acid band (1086 cm⁻¹) provided spatial discrimination between infected and uninfected cells at each time point, with balanced accuracy reaching 0.90, while point spectroscopy raised the pairwise balanced accuracy to 0.94 and confirmed the biochemical basis of the image contrast. Multiclass separation across time points reached an overall accuracy of 0.71. Single-cell heterogeneity was quantified by uniform manifold approximation and projection (UMAP), and cross-replicate reproducibility served as internal validation. These findings demonstrate that OPTIR enables label-free monitoring of viral infection in living cells using intrinsic biochemical signatures alone, providing a practical alternative to genetically encoded fluorescent reporters.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Long-time measurements of calcium release by dental resin-based composites using a liquid-contact calcium ion-selective electrode.","authors":"Paula Calli Falcoswki, Bruna Marin Fronza, Handially Dos Santos Vilela, Roberto Ruggiero Braga, Mauro Bertotti","doi":"10.1007/s00216-026-06810-w","DOIUrl":"https://doi.org/10.1007/s00216-026-06810-w","url":null,"abstract":"<p><p>Liquid-contact ion-selective electrodes have been successfully applied in electrochemical analysis over the past 60 years and remain a highly reliable platform in modern analytical chemistry. Owing to their structural simplicity, low fabrication cost, and reproducible operational behavior, these sensors offer an attractive and robust alternative for real-time measurements when their key analytical parameters are thoroughly understood. In this study, we fabricated and characterized a micro calcium ion-selective electrode (micro-ISE) based on a commercial membrane cocktail containing the ETH-1001 ionophore. Critical operational parameters were systematically evaluated to tailor the microelectrode for localized sensing, including expected measurement error, background electrolyte influence, signal stability, memory effects, and accuracy. The microsensor was subsequently positioned 20 μm above dental composite samples designed for dentin remineralization with varying inorganic compositions (100% CaHPO₄/0% glass, 50% CaHPO₄/50% glass, and 0% CaHPO₄/100% glass) to map dynamic calcium release over the first 2 h of immersion in an aqueous solution. Sensor accuracy and precision were independently validated by ICP-OES, confirming the micro-ISE's suitability for localized ion flux monitoring.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of metabolites of the epoxy resin component bisphenol A diglycidyl ether (BADGE) in human urine using gas chromatography-tandem mass spectrometry.","authors":"Jennifer Rosin, Eleonore Rissler, Pavel Dietz, Bernd Rossbach","doi":"10.1007/s00216-026-06802-w","DOIUrl":"https://doi.org/10.1007/s00216-026-06802-w","url":null,"abstract":"<p><p>Bisphenol A diglycidyl ether (BADGE) is used as an epoxy resin component in numerous applications in industry and the consumer sector. Its use for inner coatings of food and beverage cans poses a source for BADGE exposure in the general population since migration of BADGE or its derivatives from coatings into food items takes place. The compound is currently under assessment as an endocrine disruptor and a potent skin sensitiser. Nevertheless, a reliable biomarker for estimating BADGE intake by humans has yet to be identified. The objective of the study was to develop and validate a human biomonitoring method that is suitable to detect BADGE exposure down to the concentration range of environmental background exposure. The analytical method presented here enables a sensitive and specific determination of the BADGE metabolites BAGE*H₂O, BADGE*H₂O*AcOH, and BADGE*2H₂O in urine by gas chromatography-tandem mass spectrometry (GC-MS/MS). Following enzymatic hydrolysis and liquid-liquid extraction of the samples, three derivatisation steps are performed to enhance the volatility of the comparatively polar analytes. Metabolite concentrations are subsequently determined by GC-MS/MS using external calibration and isotope-labelled internal standardisation. Validation of the method yielded limits of quantification between 0.07 and 0.10 μg/L, with day-to-day precision between 4.4 and 15.2% and relative recoveries between 81 and 114% depending on the particular analyte. An analysis of n = 40 spot urine samples from the general population showed quantifiable analyte concentrations in isolated samples. Compared to the most commonly used biomarker BADGE*2H₂O, the downstream metabolite BADGE*H₂O*AcOH could be a promising alternative as it is less susceptible to contamination from external sources.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated Fe(III)-driven liquid-phase microextraction-spectrophotometry for selective and high-throughput detection of ethyl maltol in complex food samples.","authors":"Junfeng Qi, Junhe Liu, Chanjuan Hu, Qingqing Wang, Yanhong Guo, Mengdie Zhang, Li Hu, Bin Lin, Wansen Li","doi":"10.1007/s00216-026-06819-1","DOIUrl":"https://doi.org/10.1007/s00216-026-06819-1","url":null,"abstract":"<p><p>Excessive addition of ethyl maltol (EMA) to foods poses potential health risks, highlighting the need for reliable monitoring. Herein, an integrated Fe(III)-driven liquid-phase microextraction (LPME)-spectrophotometry system was developed for selective and high-throughput EMA determination. Fe(III) in the acceptor phase served dual roles: as a chemical trap achieving >90% extraction efficiency and as an in situ chromogenic probe enabling direct detection. This unique design combines membrane-mediated matrix exclusion with coordination-assisted extraction, resulting in enhanced sensitivity and improved resistance to matrix interference. Density functional theory calculations revealed that Fe(III) facilitated the forward extraction by reducing the transmembrane transfer free energy barrier while suppressing back-extraction via weakening EMA-membrane solvent interactions. Under the evaluated conditions, the system offered a broad linear range (7.8-1500 μM, R² > 0.999), low detection limit (0.67 μM), and satisfactory selectivity toward structural analogues. Validation in diverse food matrices yielded recoveries of 88-107% with good agreement with HPLC-UV (R² > 0.99). The minimal solvent consumption, high throughput, and favorable greenness further highlight its practical utility. More than an EMA detection method, this work provides a proof-of-concept for the coordination-driven LPME strategy that intrinsically integrates sample preparation with in situ signal generation, offering new insights into the design of a mechanism-driven microextraction system.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148981836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a sequential digestion workflow for dual-platform microplastic analysis in pasteurized donor human milk using µ-FTIR and Py-GC/MS.","authors":"Iffah Madiha Muhamad Shafiry, Nur Rasyiqah Shamsul, Dhea Maisarah Ahmad Nasri, Fikriah Faudzi, Hamizah Ismail, Sabiqah Tuan Anuar, Chingakham Chinglenthoiba, Norafiza Zainuddin","doi":"10.1007/s00216-026-06790-x","DOIUrl":"https://doi.org/10.1007/s00216-026-06790-x","url":null,"abstract":"<p><p>Reliable determination of microplastics in complex biological matrices remains analytically challenging because of high lipid and protein content, low particle abundance, and the risk of polymer degradation during sample preparation. This study developed and evaluated a sequential digestion and extraction workflow for isolating and characterizing microplastics in pasteurized donor human milk (PDHM). The workflow combined hexane-based lipid removal, proteinase K digestion, and controlled hydrogen peroxide treatment. Particle-count recovery was assessed using polypropylene (PP), polyethylene terephthalate (PET), and nylon 6,6 fragments measuring 0.8-1.3 mm. Recoveries ranged from 96.3% to 100%, indicating minimal loss of these fragments during digestion and filtration. The recovered fragments were analyzed by µ-FTIR. Py-GC/MS-based mass recovery was not evaluated because the dimensions of the recovered reference fragments were incompatible with the capacity of the pyrolysis sample cup, precluding their quantitative transfer and analysis. Blank-corrected microplastic abundance in PDHM ranged from 2.3 to 15.3 particles per 30 mL, equivalent to 76.7-509.9 particles L⁻¹. µ-FTIR identified polyamide (PA), polyethylene (PE), PET, polyurethane (PU), polytetrafluoroethylene (PTFE), polystyrene (PS), poly(methyl methacrylate) (PMMA), and rayon, predominantly in the <100 µm size class. Py-GC/MS analysis indicated polycarbonate at 1.5 µg per sample in two of the three replicates, whereas the remaining replicate was below the laboratory reporting limit. This finding was interpreted as preliminary because independent peak-level confirmation was not possible using the analytical information available. The developed workflow provides a basis for microplastic isolation and particle-level characterization in PDHM but requires further size- and polymer-specific validation.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148966145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient determination of ranitidine, famotidine, and cimetidine in environmental and biological matrices using a NH₂-bimetallic MIL-88(Fe/Ni)@SiO₂/Fe₃O₄ composite and dispersive micro solid-phase extraction.","authors":"Fahimeh Motamedifar, Mahboubeh Masrournia, Razieh Sanavi Khoshnood","doi":"10.1007/s00216-026-06764-z","DOIUrl":"https://doi.org/10.1007/s00216-026-06764-z","url":null,"abstract":"<p><p>The widespread use of gastrointestinal drugs like ranitidine, famotidine, and cimetidine has led to their emergence as persistent environmental contaminants, while their monitoring in biological fluids remains crucial for clinical toxicology. This study presents the development of a sensitive and efficient analytical method based on dispersive micro solid-phase extraction (d-μ-SPE) coupled with high-performance liquid chromatography-photodiode array detector (HPLC-DAD) for their determination. A novel magnetic adsorbent, NH₂-bimetallic MIL-88(Fe/Ni)@SiO₂/Fe₃O₄, was synthesized and characterized. The unique bimetallic system and amine functionalization of the metal-organic framework (MOF) provided specific interactions with the target analytes, while the silica shell ensured stability, and the magnetic core facilitated separation. The critical factors affecting the d-μ-SPE efficiency were optimized using a two-stage experimental design: a definitive screening design (DSD) identified pH, sorbent dose, desorption time, and solvent volume as significant, and a subsequent central composite design (CCD) modeled their quadratic effects. Under optimal conditions, the method demonstrated low limits of detection (0.02-0.07 ng mL⁻¹), high enrichment factors (59.3-72.5), and good precision (RSD < 5%). The method was successfully applied to tap, river, and well water, as well as human urine samples, achieving excellent relative recovery rates (89.8-95.3%), even in samples from individuals who had ingested the drugs. The adsorbent could be reused for three cycles without significant loss in performance.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of oxidized low-density lipoprotein (oxLDL)-induced inflammation in atherosclerosis using a paper-based immune-vascular model.","authors":"Praveen Kumar Gurubatha Rajadurai, Ping-Ching Pai, Kin Fong Lei","doi":"10.1007/s00216-026-06817-3","DOIUrl":"https://doi.org/10.1007/s00216-026-06817-3","url":null,"abstract":"<p><p>Atherosclerosis is driven by complex interactions between endothelial cells, smooth muscle cells, and immune cells, yet conventional in vitro models often fail to capture these multicellular dynamics. Here, we present a biomimetic paper-based immuno-vascular model that vertically integrates endothelial cells, smooth muscle cells, and THP-1-derived macrophages to simulate the vascular intima environment. Using this model, CD36-mediated inflammatory responses were investigated under oxLDL stimulation. The results revealed that CD36 expression was the highest in multicellular co-cultures, indicating synergistic effects of oxLDL and macrophage-derived paracrine signaling. The model also maintained homogeneous protein distribution and high cell viability over extended culture periods, demonstrating spatially precise assessment of immune-vascular crosstalk, including layer-to-layer propagation of inflammatory signaling consistent with translaminar paracrine communication. These immunoassay-based findings were independently corroborated within the same experimental batch by layer-separated RT-qPCR of CD36, IL-6, IL-1β, and TNF-α performed on RNA recovered directly from the destacked paper layers, which reproduced the same rank order of conditions at the transcript level. These findings highlight the critical role of intercellular signaling in potentiating inflammatory responses and validate the model as a versatile tool for studying early atherogenic processes. Beyond physiological relevance, the model offers reproducible fabrication, experimental accessibility, and compatibility with in situ molecular analyses, supporting mechanistic studies, hypothesis-driven experimentation, and early-stage therapeutic screening in cardiovascular research.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Solvent-free grinding synthesis of Hemin@P₂Mo₁₈ with peroxidase-mimicking activity for dopamine detection in bovine serum.","authors":"Shiqi Chu, Lian Yang, Zhongmei Chi, Jiahui Ma, Huanxiang Wang, Lihe Yan, Zhaoye Li, Guocheng Liu","doi":"10.1007/s00216-026-06814-6","DOIUrl":"https://doi.org/10.1007/s00216-026-06814-6","url":null,"abstract":"<p><p>Dopamine (DA) in bovine serum served as a vital physiological biomarker for evaluating livestock health status and conducting translational biomedical research. Accurate and rapid detection of DA was therefore essential for agricultural breeding monitoring and clinical biological analysis. In this work, a novel Hemin@P₂Mo₁₈ nanocomposite with peroxidase-mimicking activity was successfully prepared via a facile, green solvent-free grinding strategy. The introduction of water-soluble H₆P₂Mo₁₈ effectively suppressed the aggregation of free hemin molecules and yielded solid-phase Hemin@P₂Mo₁₈. Relative to natural peroxidases, the synthesized Hemin@P₂Mo₁₈ featured convenient recovery and excellent reusability. Given the excellent peroxidase-mimicking activity of Hemin@P₂Mo₁₈, a sensitive and selective colorimetric platform was constructed for DA detection. Moreover, the proposed approach showed high selectivity against common interfering substances and excellent stability under different conditions. It was successfully applied to the quantitative determination of DA in bovine serum samples with satisfactory recovery rates ranging from 96.8% to 100.2%. This study provided a green and efficient solvent-free grinding synthesis strategy for high-performance POM-based nanozymes. It also offered a feasible and practical approach for rapid biomarker detection in complex biological matrices, which showed great application potential in veterinary monitoring and biosensing analysis.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in the measurement of vitamin D binding protein.","authors":"Karen W Phinney, Lisa E Kilpatrick","doi":"10.1007/s00216-026-06815-5","DOIUrl":"10.1007/s00216-026-06815-5","url":null,"abstract":"<p><p>Vitamin D binding protein (DBP) is a polymorphic protein known for its function in transporting vitamin D metabolites. DBP also plays several other important roles in the body's response to tissue damage and inflammation. Measurement of DBP is fundamental to understanding its relationship to vitamin D status and to elucidating potential links between DBP concentration and various health outcomes. Early studies on DBP primarily used antibody-based techniques for quantification, and mass spectrometry-based methods have been reported during the past decade. This review will outline the different methodologies used in DBP studies and to assign values to reference materials, describe their advantages and limitations, and highlight some future trends.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13673995/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structure-guided parameter optimization of feature-based molecular networking for DIA-HRMS screening of antihistamines in cosmetics.","authors":"Guangqian Xu, Li Li, Zixuan Yang, Guiwen Guo, Siyu Peng, Jishuang Wang, Yitong Ma, Haiyan Wang","doi":"10.1007/s00216-026-06813-7","DOIUrl":"https://doi.org/10.1007/s00216-026-06813-7","url":null,"abstract":"<p><p>Feature-based molecular networking (FBMN) can support prioritization of library-absent features in high-resolution mass spectrometric screening, but parameter choices may create chemically misleading links, particularly for data-independent acquisition (DIA) spectra. We developed a structure-guided multi-metric parameter-optimization framework using 51 authenticated antihistamines assigned to structural classes independently of retention time, MS/MS similarity, and network topology. Across 595 GNPS-FBMN parameter combinations, the selected application-specific setting P264 (minimum cosine similarity, 0.475; matched ions, 8; MAX_SHIFT, 150 Da) gave an adjusted Rand index of 0.216, a same-class edge ratio of 0.697, and a cross-class edge ratio of 0.303, compared with 0.101, 0.621, and 0.379, respectively, for the controlled GNPS edge-default setting. In 120 commercial cosmetics, in-house library screening detected diphenhydramine in one sample; authentic-standard comparison confirmed the retention time, precursor ion, and MS/MS evidence, and post-confirmation measurement gave 3.28 mg kg^-1. Independent 51-target triple-quadrupole MRM analysis produced no discordant known-target result in the evaluated commercial-sample set. In a separate library-absent proof-of-concept experiment, buclizine fortified into blank cream satisfied the predefined three-anchor prioritization rule, ranked first with SIRIUS/CSI:FingerID, and was subsequently confirmed with an authentic standard. These results support the optimized FBMN as a complementary approach for structural-context assessment and library-absent candidate prioritization under the evaluated DIA-HRMS conditions.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2026-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148954724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}