Philipp Willmer, Adam C. Hundahl, Rodolphe Marie, Henrik Jensen
{"title":"Cover Picture: Continuous Titration Based Method for Rapid In-Solution Analysis of Non-Covalent Interactions (Chem. Methods 5/2025)","authors":"Philipp Willmer, Adam C. Hundahl, Rodolphe Marie, Henrik Jensen","doi":"10.1002/cmtd.202580501","DOIUrl":"10.1002/cmtd.202580501","url":null,"abstract":"<p><b>The Front Cover</b> illustrates a new method (cSPRING) to study the interaction of a target molecule exposed to a continuous titration of a small ligand inside a flow capillary. The fluorescence emission of the target is detected in two separate spectral bands. A ratiometric measurement allows to extract the binding affinity (<i>K</i><i><sub>d</sub></i>) from a single measurement. Requiring only two samples and less than a minute, this method is well-suited for high-throughput screening applications. For more details, see the Research Article by Henrik Jensen and co-workers (10.1002/cmtd.202400059).\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 5","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202580501","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143944423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Low-Micromolar Quantification of Fluorinated Analytes Using Hyperpolarized 19F Benchtop Nuclear Magnetic Resonance","authors":"Ana I. Silva Terra, Meghan E. Halse","doi":"10.1002/cmtd.202400094","DOIUrl":"10.1002/cmtd.202400094","url":null,"abstract":"<p>Benchtop nuclear magneic resonance (NMR) spectrometers are more affordable and accessible than high-field NMR instruments but suffer from higher limits of detection (LOD) and lower chemical shift resolution due to their moderate magnetic fields. Herein, reductions in the single-scan LOD values for <sup>19</sup>F benchtop (1 T) NMR of 3,5-difluoropyridine (DFP) and 2,4,6-trifluorobenzylamine (TFBA) to (6.84 ± 0.45) μM and (162 ± 28) μM are demonstrated <i>via</i> signal amplification by reversible exchange (SABRE) hyperpolarization. Further reductions in LOD to (600 ± 20) nM and (17.5 ± 2.0) μM are achieved by combining SABRE with sensitive, homogeneous, and resolved peaks in real time detection. Quantification is demonstrated over the micromolar concentration range using linear external calibration. The effects of polarization transfer to the solvent <i>via</i> proton exchange in the case of TFBA are shown to be minimized in methanol-<i>d</i><sub>4</sub>; however, <sup>19</sup>F detection and quantification are also achieved in protio methanol, with a LOD of (42.6 ± 3.6) μM.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144999059","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Thomas Julien, Boris Gouilleux, Bernard Rousseau, Michael Reggelin, Philippe Lesot
{"title":"Deuterium NMR in Chiral Polymer-Based Bimesophasic Lyotropic Mixtures: From the Demixing Process to Analytical Applications","authors":"Thomas Julien, Boris Gouilleux, Bernard Rousseau, Michael Reggelin, Philippe Lesot","doi":"10.1002/cmtd.202500011","DOIUrl":"10.1002/cmtd.202500011","url":null,"abstract":"<p>Among the order-sensitive (anisotropic) nuclear magnetic resonance (NMR) interactions, the residual <sup>2</sup>H quadrupolar couplings measured at the level of natural abundance or on deuterated compounds dissolved in (chiral) oriented solvents, is a unique and relevant source of molecular information (isotopy, chirality, structure, configuration, etc.). Until now, the aligned media used as NMR solvents (lyotropic or not) have generally been monophasic, although isotropic-anisotropic biphasic systems have recently been exploited. As a promising possibility, herein, the potential of lyotropic bimesophasic samples is examined based on a mixture of immiscible helical polymers, which result in two demixed chiral nematic domains (stable over time), each with its own orientation properties. The extraction of two independent and nonlinearly related sets of anisotropic NMR data is achieved with spatially resolved <sup>2</sup>H <i>n</i>D experiments. First applied to small achiral, prochiral and chiral deuterated compounds, the approach is extended to investigate nonenriched molecules by natural abundance deuterium 2D NMR. Here lyotropic bimesophasic chiral systems combining a polypeptide (PBLG) and a polyacetylene polymer (PLA or <span>l</span>-MSP) in organic solvents are examined in depth, in relation with several applications. The dynamic behavior of these systems (polymer demixing and exchange process) is studied by analyzing <sup>2</sup>H EXSY, <sup>2</sup>H relaxation and <sup>2</sup>H DOSY data.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 6","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202500011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Margherita Bazzoni, Yuliia Horbenko, Nour El Sabbagh, Achille Marchand, Magdalena Grochowska-Tatarczak, Aurélie Bernard, Patrick Giraudeau, François-Xavier Felpin, Jean-Nicolas Dumez
{"title":"Fast Multidimensional Flow Nuclear Magnetic Resonance at High Field for Real-Time Reaction Monitoring and Flow Synthesis","authors":"Margherita Bazzoni, Yuliia Horbenko, Nour El Sabbagh, Achille Marchand, Magdalena Grochowska-Tatarczak, Aurélie Bernard, Patrick Giraudeau, François-Xavier Felpin, Jean-Nicolas Dumez","doi":"10.1002/cmtd.202400061","DOIUrl":"10.1002/cmtd.202400061","url":null,"abstract":"<p>Flow nuclear magnetic resonance (NMR) at high field is a powerful approach for the online monitoring of chemical reactions, which provides real-time quantitative and structural information. While 1D <sup>1</sup>H spectra are commonly collected for NMR monitoring applications, their information content is limited because of peak overlap and assignment ambiguity. 2D NMR methods provide an opportunity to resolve peaks that overlap in 1D spectra and obtain correlation information that helps assignment. Their fast implementations additionally have durations (from sub-second to minute) that remain compatible with a large range of online reaction monitoring applications. They are also expected to benefit flow synthesis applications in which a flow reactor is hyphenated with a high-field NMR spectrometer. Herein, a selection of fast 2D NMR methods that are compatible with flow NMR applications is described: ultrafast 2D NMR and fast diffusion-ordered NMR spectroscopy. After a description of an experimental setup and current best practices, the principle behind each method and the way in which it can be implemented for continuously flowing samples are described. Also, the flow effects that are specific to these methods, for different settings, are described. These fast 2D NMR methods should be useful to users of flow NMR with an interest in reaction monitoring.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 6","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ekaterina V. Pokochueva, Nghia H. Le, Sylvie Guibert, Chloé Gioiosa, Quentin Stern, James Tolchard, Charlotte Bocquelet, Olivier Cala, Matthieu Cavaillès, Laurent Veyre, Otto Mankinen, Ville-Veikko Telkki, Chloé Thieuleux, Sami Jannin
{"title":"Hybrid Polarizing Solids with Extended Pore Diameters for Dissolution Dynamic Nuclear Polarization","authors":"Ekaterina V. Pokochueva, Nghia H. Le, Sylvie Guibert, Chloé Gioiosa, Quentin Stern, James Tolchard, Charlotte Bocquelet, Olivier Cala, Matthieu Cavaillès, Laurent Veyre, Otto Mankinen, Ville-Veikko Telkki, Chloé Thieuleux, Sami Jannin","doi":"10.1002/cmtd.202400068","DOIUrl":"10.1002/cmtd.202400068","url":null,"abstract":"<p>Dissolution dynamic nuclear polarization (dDNP) has emerged as a powerful technique to address the inherent sensitivity limitations in nuclear magnetic resonance imaging and spectroscopy. The technique relies on the use of stable paramagnetic polarizing agents that act as a source of hyperpolarization but unfortunately, their presence contaminates the sample and significantly enhances paramagnetic relaxation of the hyperpolarized solutions. In the last decade, novel sample formulations have been proposed to combat this issue, including silica-based hybrid polarizing solids (HYPSO), which have enabled the production of contamination-free hyperpolarization. Herein, a new generation of polarizing solids with pore diameters increased from 4 up to 28 nm is presented. Compared to the previously described HYPSOs with smaller pore sizes of 4 nm, larger pores ultimately achieve the ambitious contradictory goals of 1) providing rapid and high solid-state polarization and improving cross-polarization efficiency, while 2) decreasing hyperpolarization losses upon melting, transfer, and filtration, and 3) reducing the overall concentration of the polarizing agent by a factor of 2. These HYPSO materials can be used in virtually any existing dDNP settings without modification, apart from the sole addition of an in-line commercial filter to retain the HYPSO powder upon dissolution and transfer.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 6","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400068","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144214036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nikolett Varró, Beáta Mándityné Huszka, Eszter Erdei, András Mándoki, István M. Mándity
{"title":"Upscaled Synthesis of α- and β-Peptides in a Continuous-Flow Reactor Using Propylene Carbonate as an Eco-Friendly Solvent","authors":"Nikolett Varró, Beáta Mándityné Huszka, Eszter Erdei, András Mándoki, István M. Mándity","doi":"10.1002/cmtd.202500010","DOIUrl":"10.1002/cmtd.202500010","url":null,"abstract":"<p>A rapid, eco-friendly, and scalable continuous-flow solid-phase peptide synthesis (CF-SPPS) technology has been developed using propylene carbonate (PC) as a solvent. PC, a greener alternative to DMF, is selected based on the GSK solvent selection guide. To evaluate its compatibility with CF-SPPS, four α-peptides were synthesized, achieving high yields and purity. The method is further validated with challenging sequences, including four β-peptide foldamers and an α/β-peptide chimera. To scale up this innovative approach, larger column geometries are optimized to ensure the appropriate resin load and flow rate for desired residence times. The technology demonstrated its capability to synthesize peptides on a >4 g scale with satisfactory efficiency. A key feature is the use of PC in minimal amounts, significantly enhancing the process's green credentials. The low solvent consumption and high efficiency align with green chemistry principles, reducing waste and improving sustainability. The low process mass intensity of the method highlights its environmental benefits, making it a sustainable alternative for large-scale peptide synthesis. This breakthrough showcases the potential of CF-SPPS with PC as a solvent to meet the demands of greener and more efficient peptide production.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 8","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202500010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144869103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Claudia Adsuar Fuster, Francesco Caruso, Olivia Gómez-Laserna, Martina Romani, Ramón J. Barrio, Josep Maria Vergès, Vincenza Forgia, Maite Maguregui, Asier Vallejo
{"title":"Analytical Study of Lanolin as Potential Biomarker of Prehistoric Sheep-Shearing Practice","authors":"Claudia Adsuar Fuster, Francesco Caruso, Olivia Gómez-Laserna, Martina Romani, Ramón J. Barrio, Josep Maria Vergès, Vincenza Forgia, Maite Maguregui, Asier Vallejo","doi":"10.1002/cmtd.202400093","DOIUrl":"10.1002/cmtd.202400093","url":null,"abstract":"<p>Sheep-shearing practices probably started in the Neolithic Age when the genetic selection of sheep began to be successful in obtaining animals with abundant wool. Flint and obsidian were the main materials probably used to manufacture tools for this purpose. From shearing, lanolin wax is accumulated onto the shearing tools. Lanolin is a complex mixture of esters and polyesters, alcohols, fatty acids, and hydrocarbons produced mainly by sheep to protect the skin and the wool from environmental agents. Due to its complex chemistry, lanolin is relatively difficult to characterize. This work investigates the possible use of lanolin as a biomarker of sheep-shearing practices in the prehistoric era. For this purpose, lanolin degradation pathways have been studied by an experimental archeology approach adjuvated by Fourier transformed infrared spectroscopy, hyperspectral imaging, and gas chromatography coupled with mass spectrometry.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 8","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400093","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144869472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lanze Li, Jiexin Wen, Tsz Woon Benedict Lo, Jun Yin, Qiong Lei
{"title":"Wettability-Controlled Electrocatalytic Carbon Dioxide Reduction","authors":"Lanze Li, Jiexin Wen, Tsz Woon Benedict Lo, Jun Yin, Qiong Lei","doi":"10.1002/cmtd.202400080","DOIUrl":"10.1002/cmtd.202400080","url":null,"abstract":"<p>The electrocatalytic CO<sub>2</sub> reduction reaction (eCO<sub>2</sub>RR) offers a promising pathway for converting greenhouse gases into valuable fuels and chemicals using renewable energy. Beyond advancements in catalyst and electrolyzer design, significant opportunities lie in the strategic modulation of the gas–liquid–solid three-phase interface (TPI) on the catalyst surface. After revisiting the evolution from traditional liquid–solid double-phase interfaces to advanced gas–liquid–solid TPIs, this concept outlines major challenges in constructing stable TPIs on eCO<sub>2</sub>RR gas diffusion electrodes and reviews recent progress in TPI modulation through hydrophobicity enhancement. Further, achieving a delicate balance between hydrophobicity and hydrophilicity—optimal wettability—is crucial for optimizing TPI construction, and enhancing overall electrocatalytic performance is emphasized. This work provides valuable insights for designing efficient TPIs in eCO<sub>2</sub>RR and other gas-involved electrochemical processes, contributing to advancements in sustainable energy technologies.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 8","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400080","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144869102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Multiplex Cellular Microenvironment Detection Based on Surface-Enhanced Raman Scattering Patch","authors":"Xiaoyu Wu, Zhaodong Zhang, Feng Zhu, Yuanheng Kuang, Shuqi Chen, Chongling Sun, Bohua Liu, Yanyan Wang","doi":"10.1002/cmtd.202400083","DOIUrl":"10.1002/cmtd.202400083","url":null,"abstract":"<p>Monitoring cellular microenvironment is of vital significance to study cell process, reveal pathological state, and predict early disease. Herein, a surface-enhanced Raman scattering (SERS) patch is proposed to sense the hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) concentration and pH variation of extracellular microenvironment in a multiplex manner. Scale-up and ordered gold nanorods (AuNRs) assembly is introduced by interfacial assembly and absorption transferring on the flexible polydimethylsiloxane (PDMS) film, providing the patch with great SERS enhancement for biosensing. The modification of H<sub>2</sub>O<sub>2</sub>-responsive and pH-susceptive Raman reporter molecule endows the SERS patch with high sensitivity to multiple analytes, and the construction of multivariate curve resolution (MCR) analysis facilitates the quantification of H<sub>2</sub>O<sub>2</sub> and pH value in spite of the characteristic peaks overlap. Furthermore, the SERS patch is used to compare the extracellular microenvironments of different living cells. Results demonstrate that highly migratory cells (i.e., MDA-MB-231) secreted more H<sub>2</sub>O<sub>2</sub> under phorbol 12-myristate 13-acetate (PMA) stimulation, and the extracellular microenvironment is more acidic than the cells with weak migration ability (i.e., MCF-7), revealing the relationship between cell migration and microenvironment. The strategy integrates H<sub>2</sub>O<sub>2</sub> detection and pH determination in one patch, which may offer an approach for novel multiplex biosensor and cell migration investigation.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 7","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400083","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144615289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Siwon Yun, Hyeji Han, Muhammad Adnan, Wonjong Lee, Min Kim, Jongchul Lim, Jinseck Kim
{"title":"Electron Transport Bilayer Design for Perovskite-Based Tandem Solar Cells Using Thermal Evaporation","authors":"Siwon Yun, Hyeji Han, Muhammad Adnan, Wonjong Lee, Min Kim, Jongchul Lim, Jinseck Kim","doi":"10.1002/cmtd.202500015","DOIUrl":"https://doi.org/10.1002/cmtd.202500015","url":null,"abstract":"<p>The development of high-performance perovskite solar cells (PSCs) and perovskite-based tandem solar cells (PTSCs) has attracted significant attention from researchers owing to their simple and cost-effective fabrication process. However, careful consideration is required regarding the selection of charge-transport layers, which play a key role in improving the efficiency and stability of devices. In p–i–n (inverted) PSCs, fullerene and its derivatives are widely employed as electron-transport layers (ETLs). Among them, fullerene is compatible with low-temperature and vacuum-based solar cell fabrication processes. However, the single layer of fullerene tends to form nonuniform layers due to its aggregation propensity. Furthermore, it struggles to withstand the damage that occurs during the formation of transparent conductive layers in tandem configurations. To overcome the limitation of fullerene, employing bilayer ETL has emerged as a promising strategy. Therefore, in this study, the evaporation processes for ETLs in inverted PSCs are focused on, emphasizing the importance of bilayer ETL architectures in enhancing the efficiency and durability of PTSCs. The discussion highlights material options for bilayer ETLs and examines optimal thickness parameters to achieve superior efficiency. By providing a comprehensive understanding of bilayer ETLs, this review aims to guide future advancements in the PSCs and PTSCs technologies.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 10","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202500015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145243170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}