Advances in Sample Preparation最新文献

{"title":"A green approach for electromembrane extraction of morphine from urine using sodium alginate-g-polyacrylic acid/agarose hydrogel membrane","authors":"Elnaz Khodabakhshi ,&nbsp;Somayeh Makarem ,&nbsp;Mehran Kurdtabar ,&nbsp;Saeed Nojavan","doi":"10.1016/j.sampre.2025.100204","DOIUrl":"10.1016/j.sampre.2025.100204","url":null,"abstract":"<div><div>The development of novel membranes through green chemistry remains a significant challenge in advancing practical separation science. In this study, a sodium alginate-g-polyacrylic acid/agarose hydrogel was synthesized and, for the first time, applied as a membrane in gel electromembrane extraction (G-EME) for the isolation of morphine, a basic analyte, from urine samples. The extracted morphine was subsequently quantified using differential pulse voltammetry (DPV) with a glassy carbon electrode. The hydrogel membrane (5 mm thick) was prepared using 0.75 % (w/v) sodium alginate, 3.80 % (v/v) acrylic acid, 0.04 % (w/v) ammonium persulfate, and 1.25 % (w/v) agarose. The extraction process was optimized by evaluating factors such as membrane composition, extraction time, applied voltage, and pHs of the sample solution, membrane, and acceptor phase (AP). Under optimal conditions (25 min extraction time, 70 V applied voltage, membrane pH 4.0, AP pH 3.0, and sample solution pH 7.0) morphine was efficiently extracted through the hydrogel membrane and transferred to an aqueous AP. The collected AP was mixed with 0.15 M phosphate buffer (pH 7.0) and analyzed voltammetrically. The method achieved a limit of detection of 0.09 μg/mL and a limit of quantification of 0.28 μg/mL, and was successfully applied for morphine determination in urine sample. The polyacrylic acid-based membrane enhanced extraction efficiency through electrostatic interactions, while the use of DPV offered clearer signal interpretation and reduced background current, facilitating reliable detection of trace analytes. These advantages support the method’s potential for simple, selective, and environmentally friendly analysis of basic drugs in biological samples.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"15 ","pages":"Article 100204"},"PeriodicalIF":6.5,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144780679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improved sample preparation using bead mill homogenization","authors":"Madison E. Walbrun ,&nbsp;Aamna Aijaz ,&nbsp;Gabriella Ryan ,&nbsp;John D. Bunting ,&nbsp;Zachary P. Morehouse ,&nbsp;Rodney J. Nash","doi":"10.1016/j.sampre.2025.100194","DOIUrl":"10.1016/j.sampre.2025.100194","url":null,"abstract":"<div><div>Proper sample preparation is the key to having successful data analysis downstream. Despite the many efforts to improve sample preparation, there are still many inconsistencies and challenges when attempting to produce a sample for downstream analysis. Here we compare common methods used in sample preparation (detergent and manual Dounce homogenization) to mechanical Bead Mill homogenization. These methods have been shown to lyse both cell and tissue, however, concentrations and detectability of desired analytes can be greatly reduced. These results show that mechanical Bead Mill homogenization requires less starting sample, and less time, while producing higher protein concentrations with increased reproducibility, therefore improving downstream analysis.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"15 ","pages":"Article 100194"},"PeriodicalIF":5.2,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144556795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of performance and matrix compatibility of mixed mode C18-SCX SPME fibers for compounds with different physicochemical properties","authors":"Marcos Tascon ,&nbsp;Ezel Boyaci ,&nbsp;Nathaly Reyes-Garcés ,&nbsp;Janusz Pawliszyn","doi":"10.1016/j.sampre.2025.100195","DOIUrl":"10.1016/j.sampre.2025.100195","url":null,"abstract":"<div><div>Solid-phase microextraction (SPME) has evolved significantly since its inception, yet challenges remain in developing coatings fully compatible with LC-MS that combine broad polarity coverage with biocompatibility for complex matrices. This study evaluates mixed-mode C₁₈-SCX (strong cation exchange) SPME fibers designed to extract analytes of a wide range of physicochemical properties, addressing limitations in current methodologies. The fibers were tested for extraction efficiency, reproducibility, and matrix compatibility using a group of model compounds with different physicochemical properties, namely, codeine (logP=1.19), carbamazepine (logP=2.45), diazepam (logP=2.82), and propranolol (logP=3.47). Furthermore, the biocompatibility was tested in diverse matrices, such as PBS, blood, plasma, urine, and grape juice.</div><div>Results demonstrated exceptional inter-fiber reproducibility (RSD ≤ 15 %, <em>n</em> = 96 fibers) and robust performance in biomatrices, with relative matrix effects primarily governed by analyte binding affinities to matrix macromolecules rather than coating fouling. Absolute matrix effects were negligible (93–111 %), underscoring the fibers’ ability to deliver clean extracts for LC-MS analysis. Fiber reusability was validated over three consecutive extractions (RSD ≤ 10 %), and morphological integrity was preserved post-extraction, even in challenging matrices like whole blood. This work represents the versatility of mixed-mode SPME fibers for high-throughput bioanalysis, offering a significant advancement for in vivo and in vitro targeted and untargeted applications.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"15 ","pages":"Article 100195"},"PeriodicalIF":5.2,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144549565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A 3D-printed handheld device for quick citrus tissue lysis and nucleic acid extraction","authors":"Chia-Wei Liu ,&nbsp;Brent Kalish ,&nbsp;Sohrab Bodaghi ,&nbsp;Georgios Vidalakis ,&nbsp;Hideaki Tsutsui","doi":"10.1016/j.sampre.2025.100199","DOIUrl":"10.1016/j.sampre.2025.100199","url":null,"abstract":"<div><div>A 3D-printed handheld device has been developed for rapid and efficient sample preparation, extracting pathogen total nucleic acids from citrus leaves for downstream molecular analysis. With its high-speed motor, knurled lysis chamber for rapid sample lysis, and quick nucleic acid extraction using cellulose paper disks (Whatman CHR1), this device can yield ready-to-use DNA and RNA in just 12 min. All components, except for the motor, wiring, and paper disks, were printed in-house using a PolyJet 3D printer with photosensitive resins. The device was optimized for maximum sample lysis by evaluating operation voltages and chamber features, as measured by DNA and RNA concentrations using a Qubit fluorometer and quantification cycle (Cq) values obtained through qPCR assays. The results showed that the lysis chamber with internal knurling and the motor operated at 7.5 V was sufficient for effective sample lysis in 1 min, achieving RNA concentrations up to 87.6 % of those obtained with mortar-and-pestle grinding. Paper disk washing and elution conditions were also optimized using a NanoDrop spectrophotometer and qPCR assays, where two 1-minute washes and a 100 µL elution volume resulted in the highest purity and lowest Cq value. The optimized handheld device was validated with citrus sources infected with <em>Citrus tristeza virus</em> (CTV) and <em>Spiroplasma citri</em> (<em>S. citri</em>), demonstrating high detection accuracy (100 %) and low assay variation (CV &lt; 3.8 %) in qPCR-based analysis. Based on these successful results, this device is expected to be broadly applicable to similar viral and bacterial pathogens affecting plants.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"15 ","pages":"Article 100199"},"PeriodicalIF":5.2,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144634324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"3D-printed device for improved membrane-based extraction procedure of xenobiotics in complex matrices","authors":"Miryam Perrucci ,&nbsp;Erika Maria Ricci ,&nbsp;Abuzar Kabir ,&nbsp;Kenneth G. Furton ,&nbsp;Marcello Locatelli","doi":"10.1016/j.sampre.2025.100205","DOIUrl":"10.1016/j.sampre.2025.100205","url":null,"abstract":"<div><div>To minimize the environmental and human health impacts of chemical processes, there is growing interest in eco-friendly methods aligned with the principles of Green Analytical Chemistry (GAC). A successful chemical analysis typically involves sample preparation, sampling, separation and analysis, quantification, and data interpretation. Among these, sample preparation plays a crucial role in isolating and preconcentrating target analytes from complex matrices.</div><div>In 2014, fabric phase sorptive extraction (FPSE) was introduced as a simplified and greener sample pretreatment method. Building on this advancement and a recent patent, the present study introduces an innovative 3D-printed device designed for use with a range of target analytes and complex sample matrices. Constructed from inert materials, the device features a rigid, modular structure with multiple windows that securely hold various membrane-based extraction materials. These include FPSE membranes, electrospun membranes, and materials derived from adsorbent systems recovered from production or usage waste, as well as permeable molecularly imprinted polymers (MIPs).</div><div>The device offers several advantages, including enhanced enrichment factors, compatibility with diverse planar membrane types, and highly customizable selectivity based on membrane configuration and chemistry. Its design also incorporates a built-in slot for a magnetic stirrer, enabling precise control of rotation speed during extraction-even <em>in field</em> conditions using a portable, battery-powered stirrer.</div><div>Experimental results unequivocally demonstrate that the new device achieves superior enrichment factors compared to previously validated methods for the same analytes, confirming its effectiveness and potential for broader analytical applications.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"15 ","pages":"Article 100205"},"PeriodicalIF":6.5,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144780678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advances in environmental sample preparation: Nanomaterials, automation, and sustainable strategies","authors":"Federico Belén ,&nbsp;Julián Gutiérrez ,&nbsp;Valeria Springer","doi":"10.1016/j.sampre.2025.100171","DOIUrl":"10.1016/j.sampre.2025.100171","url":null,"abstract":"<div><div>The increasing occurrence of micropollutants and the need for their monitoring in environmental matrices have driven the demand for more efficient and accurate analytical methods. In particular, sample preparation procedures are essential for addressing the complexity of environmental samples while ensuring high sensitivity. To meet these demands, various nanomaterials (NMs) have been proposed as extractive phases for development of miniaturized sorbent-based extraction approaches. These NMs, ranging from carbon-based nanostructures and metal-based or metal oxide nanoparticles, to metal–organic composites, are at the forefront of innovation due to their exceptional surface areas, tunable properties, ease of functionalization and, in some cases, green production routes. On the other hand, the development of (semi)automated platforms has facilitated high-throughput and reproducible sample processing, significantly reducing the use of reagents, time, and labor. This review highlights the pivotal role of NMs and automation in advancing sample preparation methodologies over the past five years, in alignment with green sample preparation (GSP) guidelines. Additionally, the integration of emerging technologies, such as 3D printing, is discussed in the context of developing modern sample preparation methods. Recent contributions involving miniaturized platforms, green nanosorbents, and the integration of processing steps with automation, are evaluated for their potential to provide highly efficient, cost-effective solutions for monitoring micropollutants in environmental matrices.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"14 ","pages":"Article 100171"},"PeriodicalIF":5.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143619453","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":"Grapefruit peel activated carbon for multi-analyte microextraction in water analysis through rotating disk sorptive extraction","authors":"Alejandra Molina-Balmaceda,&nbsp;Valentina Rojas-Candia,&nbsp;Juan José Triviño,&nbsp;Daniel Arismendi,&nbsp;Pablo Richter","doi":"10.1016/j.sampre.2025.100176","DOIUrl":"10.1016/j.sampre.2025.100176","url":null,"abstract":"<div><div>In the realm of green analytical chemistry, there is growing interest in sorbent phases derived from natural materials like grapefruit peels as promising alternatives to commercial sorbent phases. Initially, the effectiveness of grapefruit peels in their natural state was compared to activated carbons treated with activating agents as a sorbent phase in rotating disk sorptive extraction (RDSE). The conditions for synthesizing activated carbon were optimized by adjusting the activating agent variables, proportions, and temperatures to obtain an efficient material for extracting ethylparaben (EP), propylparaben (PP), ibuprofen (Ibu), triclosan (TCS), bisphenol A (BPA), and 17-α-ethinylestradiol (EE2) using rotating disk sorption extraction technique from aqueous samples. The optimal conditions were determined to be activation with ZnCl₂ at a 1:1.2 ratio (material: activating reagent) and carbonization at 400 °C (AC400(Z1.2)). Characterization revealed a hydrophilic microporous material with a large surface area and aromatic structure, confirming its potential as a sorbent phase for aqueous sample preparation. The application of AC400(Z1.2) sorbent phase in RDSE in river water confirmed its effectiveness, revealing EP, PP, Ibu, TCS, and BPA in concentrations ranging from 0.18 to 2.8 µg <span>l</span>^-1, with EE2 concentration below the limit of detection. Reusability studies demonstrate that this material can be reused for the simultaneous extraction of analytes in at least two consecutive extractions without requiring additional treatment between extractions. This material proves to be an economical and bio-based alternative to commercial sorbent phases. Finally, its application in RDSE was evaluated using green analytical chemistry metrics (AGREEprep and BAGI), resulting in a methodology with a low environmental impact.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"14 ","pages":"Article 100176"},"PeriodicalIF":5.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143681877","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":"Sustainable analytical approaches for microplastics in wastewater, sludge, and landfills: Challenges, fate, and green chemistry perspectives","authors":"Maryam Mallek ,&nbsp;Damia Barcelo","doi":"10.1016/j.sampre.2025.100178","DOIUrl":"10.1016/j.sampre.2025.100178","url":null,"abstract":"<div><div>The effects of microplastics (MPs) in wastewater, sludge, and landfills are analyzed and discussed, focusing on their pathways into the environment. The application of sludge as a soil amendment and the reuse of treated wastewater in agriculture have been further proven to contribute to the introduction of MPs into terrestrial ecosystems. MPs present a capacity to accumulate in plants together with co-contaminants such as heavy metals and pharmaceuticals, posing additional risks to food safety and soil quality. This review underscores the need to align analytical practices with sustainability by adopting environmentally friendly methods. A critical discussion is presented on the analysis of MPs, encompassing an in-depth examination of analytical methodologies, challenges in sample preparation, and potential solutions. Techniques such as Fourier Transform Infrared Spectroscopy (FTIR), Raman Spectroscopy, and Pyrolysis-GC–MS (Py-GC–MS) are highlighted for their effectiveness in MP identification, combined with treatment approaches like density separation, enzymatic digestion, and oxidation. MPs appear in various forms, including synthetic fibers, microbeads, and fragmented particles, with their distribution being influenced by polymer types. Low-density polymers like polyethylene (PE) and polypropylene (PP) tend to remain in the water phase, whereas high-density polymers such as polyester (PES/PEST) and polytetrafluoroethylene (PTFE) accumulate in solid matrices like sludge and sediments. Inconsistencies in measurement units are a significant issue in microplastic analysis. Concentrations are typically reported as items per liter (for water) or items per kilogram (for soil), However, mass-based units such as µg/L are rarely employed, complicating data comparison. Another essential component is the determination of MP size, which is largely dependent on analytical equipment detection limits and resolution. Most studies focus on MPs ranging from 20–50 µm to a few millimeters, with limited research addressing MPs smaller than 0.1 µm, known as nanoplastics (NPs). Lastly, sustainable management approaches for addressing the worldwide problem of plastic pollution will be presented.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"14 ","pages":"Article 100178"},"PeriodicalIF":5.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143739260","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":"Microextraction by packed sorbent for the determination of selected synthetic cathinones and 2C-P in hair","authors":"Ana Y. Simão ,&nbsp;Luana M. Rosendo ,&nbsp;Pedro Dinis ,&nbsp;Cláudia Margalho ,&nbsp;Maristela Andraus ,&nbsp;Mário Barroso ,&nbsp;Eugenia Gallardo","doi":"10.1016/j.sampre.2025.100175","DOIUrl":"10.1016/j.sampre.2025.100175","url":null,"abstract":"<div><div>New psychoactive substances (NPS), including synthetic cathinones and phenethylamines, pose significant challenges due to their evolving chemical structures and health impacts. This study aimed to optimize and validate a methodology for detecting and quantifying a number of synthetic cathinones [methylone, ethylone, pentedrone, 4-chloroethcathinone (4-CEC), penthylone, α-pyrrolidinopentiophenone (α-PVP), 3,4-methylenedioxypyrovalerone (MDPV), 4‑chloro-α-pyrrolidinovalerophenone (4-Cl-α-PVP), as well as phenethylamine 2,5-dimethoxy-4(n)-propylphenethylamine (2C-P)], in hair samples using microextraction by packed sorbent (MEPS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). We developed an environmentally friendly and cost-effective MEPS procedure, which was validated for selectivity, linearity, precision, accuracy, and recovery. Results demonstrated that the method effectively detects analytes with limits of quantification as low as 10 pg/mg. The method successfully identifies MDPV in concentrations consistent with those observed in chronic and acute drug consumption cases, proving its suitability in forensic and clinical toxicology. This approach offers a robust solution for forensic and clinical applications, by combining efficient sample clean-up with precise analytical capabilities.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"14 ","pages":"Article 100175"},"PeriodicalIF":5.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143643202","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":"On the greenness of recent microwave-assisted digestion methods: An evaluation with AGREEprep","authors":"Juan L. Benedé ,&nbsp;Francisco Pena-Pereira","doi":"10.1016/j.sampre.2025.100177","DOIUrl":"10.1016/j.sampre.2025.100177","url":null,"abstract":"<div><div>Microwave-assisted digestion (MAD) is a widely used technique for the preparation of samples for elemental analysis. Significant advances toward more efficient and less hazardous MAD methods have been described in recent years, even though no systematic evaluation has been carried out in terms of green sample preparation. The present work evaluates the effectiveness and environmental impact of recent MAD methods through the lens of AGREEprep, a recognized metric tool that identifies the greenness attributes of sample preparation methods. The metric tool was employed for the assessment of 38 decomposition methods for food analysis, including MAD methods involving decreasing amounts of oxidizing agents, considering supplemental reagents and assistances, such as oxygen pressure, hydrogen peroxide, ultraviolet irradiation, and microwave-induced combustion. In addition, conventional decomposition methods and alternative approaches involving infrared heating have been considered in the assessment. The 38 methods received overall scores in a wide range of 0.14 to 0.52, being MAD methods without acid consumption and those involving combustion the greenest methods (scores from 0.40 to 0.52). By systematically evaluating key criteria, such as reagent usage, energy consumption, process safety, and waste generation, AGREEprep provides a comprehensive view of MAD methods that can contribute to further improvements toward more sustainable analytical practices.</div></div>","PeriodicalId":100052,"journal":{"name":"Advances in Sample Preparation","volume":"14 ","pages":"Article 100177"},"PeriodicalIF":5.2,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143681937","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}