Ava Rossetti, xenia kostoulias, Magdalena Giergiel, Jhih-Hang Jiang, Anton Y Peleg, Kamila Kochan
Fast diagnosis of antimicrobial resistance (AMR), especially in high-priority pathogens, is essential to maintain the best-possible patient outcomes. In this communication, we present a new approach for rapid AMR assessment that is able to distinguish between vancomycin-resistant and vancomycin-susceptible enterococci (VRE and VSE, respectively) within 2 hours using ATR-FTIR spectroscopy, chemometrics and chemometrics-based modelling. This study outlined how early spectral markers of effective vancomycin action were used to accurately predict resistance profiles in both known and blinded Enterococcus samples. This paper provides the foundation for a rapid diagnostic tool that enables faster determination of bacterial susceptibility, supporting timely clinical decisions and better patient outcomes.
{"title":"Vibrational spectroscopy for rapid profiling of vancomycin susceptibility in Enterococci","authors":"Ava Rossetti, xenia kostoulias, Magdalena Giergiel, Jhih-Hang Jiang, Anton Y Peleg, Kamila Kochan","doi":"10.1039/d5an01068c","DOIUrl":"https://doi.org/10.1039/d5an01068c","url":null,"abstract":"Fast diagnosis of antimicrobial resistance (AMR), especially in high-priority pathogens, is essential to maintain the best-possible patient outcomes. In this communication, we present a new approach for rapid AMR assessment that is able to distinguish between vancomycin-resistant and vancomycin-susceptible enterococci (VRE and VSE, respectively) within 2 hours using ATR-FTIR spectroscopy, chemometrics and chemometrics-based modelling. This study outlined how early spectral markers of effective vancomycin action were used to accurately predict resistance profiles in both known and blinded <em>Enterococcus</em> samples. This paper provides the foundation for a rapid diagnostic tool that enables faster determination of bacterial susceptibility, supporting timely clinical decisions and better patient outcomes.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"103 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146070162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maria Clara D N G Leal,Binhan Yu,Tianzhi Wang,Junji Iwahara
Advances in probe hardware that can produce strong field gradients have expanded the applicability of NMR-based diffusion measurements for various inorganic ions (e.g., Na+, K+, Li+, Mg2+, Cl-, and SO42-), thereby facilitating a wide range of chemical research. However, as we demonstrate in this paper, the commonly used NMR pulse sequences for diffusion measurements are not optimally suited for inorganic ions. For many inorganic ions, their NMR-active nuclei are quadrupolar nuclei with small gyromagnetic ratios and rapid longitudinal relaxation. These properties inherently reduce sensitivity, particularly when magnetization must be stored along the longitudinal axis, as required in stimulated-echo schemes. Strong gradients also demand efficient suppression of eddy current effects. Using 39K and 25Mg diffusion NMR experiments on DNA solutions, we demonstrate that the bipolar-pair (BPP) spin-echo method provides more than a 2-fold improvement in sensitivity compared to the BPP stimulated-echo method. While both methods yield consistent diffusion coefficients and achieve equally effective suppression of eddy currents, the BPP spin-echo method provides notably improved precision due to its higher signal-to-noise ratio in the NMR spectra. When strong gradients are available, the BPP spin-echo method is a more robust and sensitive option for diffusion NMR of inorganic ions, reducing the measurement time by a factor of 4 compared to the stimulated-echo method.
{"title":"Improved NMR-based diffusion measurements for inorganic ions.","authors":"Maria Clara D N G Leal,Binhan Yu,Tianzhi Wang,Junji Iwahara","doi":"10.1039/d5an01232e","DOIUrl":"https://doi.org/10.1039/d5an01232e","url":null,"abstract":"Advances in probe hardware that can produce strong field gradients have expanded the applicability of NMR-based diffusion measurements for various inorganic ions (e.g., Na+, K+, Li+, Mg2+, Cl-, and SO42-), thereby facilitating a wide range of chemical research. However, as we demonstrate in this paper, the commonly used NMR pulse sequences for diffusion measurements are not optimally suited for inorganic ions. For many inorganic ions, their NMR-active nuclei are quadrupolar nuclei with small gyromagnetic ratios and rapid longitudinal relaxation. These properties inherently reduce sensitivity, particularly when magnetization must be stored along the longitudinal axis, as required in stimulated-echo schemes. Strong gradients also demand efficient suppression of eddy current effects. Using 39K and 25Mg diffusion NMR experiments on DNA solutions, we demonstrate that the bipolar-pair (BPP) spin-echo method provides more than a 2-fold improvement in sensitivity compared to the BPP stimulated-echo method. While both methods yield consistent diffusion coefficients and achieve equally effective suppression of eddy currents, the BPP spin-echo method provides notably improved precision due to its higher signal-to-noise ratio in the NMR spectra. When strong gradients are available, the BPP spin-echo method is a more robust and sensitive option for diffusion NMR of inorganic ions, reducing the measurement time by a factor of 4 compared to the stimulated-echo method.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"77 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146056941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
MXene-based fluorescent aptasensors leverage the synergistic integration of the intrinsic physicochemical properties of MXenes, including tunable surface chemistry, broad-spectrum optical absorption, and superior fluorescence quenching efficiency, with the molecular recognition capabilities and strong binding affinity of aptamers. These two-dimensional transition metal carbides and nitrides efficiently suppress background fluorescence in dye-labeled aptamer systems through electrostatic interactions and π-π stacking. In the absence of the target analyte, the aptamers adsorb onto the MXene surface, facilitating non-radiative energy transfer and thereby suppressing the signal. Upon specific target recognition, a conformational rearrangement of the aptamer reduces its surface affinity, leading to desorption and subsequent fluorescence recovery via a target-induced "signal-on" mechanism. Such platforms demonstrate ultra-low detection limits, excellent selectivity, and modular adaptability for the detection of a broad spectrum of analytes, including clinical biomarkers, pathogenic microorganisms, environmental toxins, and heavy metal ions. This comprehensive review systematically summarises the mechanistic foundations of MXene-aptamer interactions, recent advancements in analytical applications, and emerging directions for translational development in biomedical diagnostics and environmental monitoring.
{"title":"MXene-based fluorescent aptasensors: advances and prospects in diagnostics and environmental monitoring.","authors":"Rajapriya Govindaraju,Jongsung Kim","doi":"10.1039/d5an00945f","DOIUrl":"https://doi.org/10.1039/d5an00945f","url":null,"abstract":"MXene-based fluorescent aptasensors leverage the synergistic integration of the intrinsic physicochemical properties of MXenes, including tunable surface chemistry, broad-spectrum optical absorption, and superior fluorescence quenching efficiency, with the molecular recognition capabilities and strong binding affinity of aptamers. These two-dimensional transition metal carbides and nitrides efficiently suppress background fluorescence in dye-labeled aptamer systems through electrostatic interactions and π-π stacking. In the absence of the target analyte, the aptamers adsorb onto the MXene surface, facilitating non-radiative energy transfer and thereby suppressing the signal. Upon specific target recognition, a conformational rearrangement of the aptamer reduces its surface affinity, leading to desorption and subsequent fluorescence recovery via a target-induced \"signal-on\" mechanism. Such platforms demonstrate ultra-low detection limits, excellent selectivity, and modular adaptability for the detection of a broad spectrum of analytes, including clinical biomarkers, pathogenic microorganisms, environmental toxins, and heavy metal ions. This comprehensive review systematically summarises the mechanistic foundations of MXene-aptamer interactions, recent advancements in analytical applications, and emerging directions for translational development in biomedical diagnostics and environmental monitoring.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"102 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146056943","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Though not very popularly recognised as a pathogenic disease, yet each year silently the morbidity and mortality of Leptospirosis has rapidly increased up to 1.03 million and 58,900 respectively. The zoonotic property of Leptospirosis further escalates concerns regarding the stability of human and one health. Hence, early and specific detection for the prompt initiation of treatment becomes exceedingly imperative. In the present article, various conventional and molecular methods including microscopic agglutination test (MAT), enzyme linked immunosorbent assays (ELISA), polymerase chain reaction (PCR), loop mediated isothermal amplification (LAMP), Mass spectrometry (MS) and rapid diagnostic tests have been comprehensively and critically reviewed for the detection of Leptospirosis based on the available literatures. This review is the first of its kind in consolidating the mass spectrometric methods that have been employed and the limitations and achievements of these sophistications have also been highlighted. The review disclosed that analytical expertise available has not been fully utilized in case of Leptospira detection. Future directions and recommendations have been put forth for rapid, simple, sensitive, accurate and early detection of Leptospira.
{"title":"Interrogating the existing analytical palette deployed for Leptospirosis detection and diagnostics","authors":"Prasanna Kannan Embar, Prasanth Venkatachalam, Judy Gopal, Nazim Hasan, Manikandan Muthu","doi":"10.1039/d6an00005c","DOIUrl":"https://doi.org/10.1039/d6an00005c","url":null,"abstract":"Though not very popularly recognised as a pathogenic disease, yet each year silently the morbidity and mortality of Leptospirosis has rapidly increased up to 1.03 million and 58,900 respectively. The zoonotic property of Leptospirosis further escalates concerns regarding the stability of human and one health. Hence, early and specific detection for the prompt initiation of treatment becomes exceedingly imperative. In the present article, various conventional and molecular methods including microscopic agglutination test (MAT), enzyme linked immunosorbent assays (ELISA), polymerase chain reaction (PCR), loop mediated isothermal amplification (LAMP), Mass spectrometry (MS) and rapid diagnostic tests have been comprehensively and critically reviewed for the detection of Leptospirosis based on the available literatures. This review is the first of its kind in consolidating the mass spectrometric methods that have been employed and the limitations and achievements of these sophistications have also been highlighted. The review disclosed that analytical expertise available has not been fully utilized in case of Leptospira detection. Future directions and recommendations have been put forth for rapid, simple, sensitive, accurate and early detection of Leptospira.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"40 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146089616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ocean acidification (OA) refers to the ongoing decline in ocean pH caused by the cascading effects of increased atmospheric CO2, which has significant negative impacts on various marine organisms, particularly crustaceans with calcified shells. However, research on the metabolic responses of crustaceans remains limited. In this study, we performed untargeted metabolomics on hemolymph samples from Cancer borealis (Jonah crab), a crustacean species well known for its tolerance to temperature and pH changes, to investigate its metabolic responses to OA. Two extraction methods—isopropanol (IPA) and acidified methanol (AcMeOH)—were employed to capture a broad range of metabolites and small peptides. Both methods enabled comprehensive detection; however, IPA yielded more consistent and extensive metabolite coverage, identifying 43 lipids compared to only 15 with AcMeOH. We identified 15 metabolites that responded significantly to OA. Several metabolites, including the potential neuropeptide cycloprolylglycine and the exogenous compound curcumin, exhibited concentration changes under OA exposure, suggesting their potential relevance in stress response pathways triggered by environmental stress. Overall, we highlight IPA as a more effective extraction method for untargeted metabolomics in crustacean hemolymph. Our study elucidates metabolic dynamics that enhance our understanding of the physiological adaptability of marine crustaceans under environmental stress and provides a comprehensive dataset that for future OA research.
{"title":"Untargeted Mass Spectrometry to Investigate Ocean Acidification in Cancer borealis Using Optimized Metabolite Extraction methods","authors":"Yunxiao Yao, Olga Riusech, Shuling XU, Lingjun Li","doi":"10.1039/d5an00788g","DOIUrl":"https://doi.org/10.1039/d5an00788g","url":null,"abstract":"Ocean acidification (OA) refers to the ongoing decline in ocean pH caused by the cascading effects of increased atmospheric CO2, which has significant negative impacts on various marine organisms, particularly crustaceans with calcified shells. However, research on the metabolic responses of crustaceans remains limited. In this study, we performed untargeted metabolomics on hemolymph samples from Cancer borealis (Jonah crab), a crustacean species well known for its tolerance to temperature and pH changes, to investigate its metabolic responses to OA. Two extraction methods—isopropanol (IPA) and acidified methanol (AcMeOH)—were employed to capture a broad range of metabolites and small peptides. Both methods enabled comprehensive detection; however, IPA yielded more consistent and extensive metabolite coverage, identifying 43 lipids compared to only 15 with AcMeOH. We identified 15 metabolites that responded significantly to OA. Several metabolites, including the potential neuropeptide cycloprolylglycine and the exogenous compound curcumin, exhibited concentration changes under OA exposure, suggesting their potential relevance in stress response pathways triggered by environmental stress. Overall, we highlight IPA as a more effective extraction method for untargeted metabolomics in crustacean hemolymph. Our study elucidates metabolic dynamics that enhance our understanding of the physiological adaptability of marine crustaceans under environmental stress and provides a comprehensive dataset that for future OA research.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"73 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146057107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ambient ionization sources enable analysis via mass spectrometry (MS) with minimal sample handling and without vacuum-based ionization/sampling. The versatile nature of ambient ionization techniques makes them well suited for both high-throughput analyses and in situ spatial characterization. Ambient MS platforms such as desorption electrospray ionization (DESI-MS), direct analysis in real time (DART-MS), paper spray (PS-MS), and secondary electrospray ionization (SESI-MS) are particularly amenable for microbial analysis and have recently been utilized for rapid profiling of microorganisms and imaging of fragile substrates with complex biochemistries. This minireview aims to provide an overview of contemporary ambient ionization technologies coupled with MS and summarize the recent application areas of these strategies in the characterization of microbial systems via mass spectrometry over the past five years.
{"title":"Ambient ionization strategies for the characterization of microbial systems via mass spectrometry","authors":"Hawkins S. Shepard, Jody C. May, John A. McLean","doi":"10.1039/d5an01214g","DOIUrl":"https://doi.org/10.1039/d5an01214g","url":null,"abstract":"Ambient ionization sources enable analysis <em>via</em> mass spectrometry (MS) with minimal sample handling and without vacuum-based ionization/sampling. The versatile nature of ambient ionization techniques makes them well suited for both high-throughput analyses and <em>in situ</em> spatial characterization. Ambient MS platforms such as desorption electrospray ionization (DESI-MS), direct analysis in real time (DART-MS), paper spray (PS-MS), and secondary electrospray ionization (SESI-MS) are particularly amenable for microbial analysis and have recently been utilized for rapid profiling of microorganisms and imaging of fragile substrates with complex biochemistries. This minireview aims to provide an overview of contemporary ambient ionization technologies coupled with MS and summarize the recent application areas of these strategies in the characterization of microbial systems <em>via</em> mass spectrometry over the past five years.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"41 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146048797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Molecular beacons offer a versatile platform for detecting specific nucleic acid targets based on the relative proximity of fluorophore and quencher molecules. Conventional beacons contain a single target-binding site in the loop, and hybridization has been modeled based on three specific phases: closed hairpin, random coil, and target-bound. The target-beacon interactions can be tuned by stem length and target site location, while multi-site beacons introduce allosteric effects that modulate affinity and produce steeper, switch-like responses. Despite available thermodynamic models, current in silico methods do not adequately predict complex sensor outputs, especially for multi-site interactions. We present a new mathematical framework for calculating the thermodynamic parameters of beacons with two target binding sites. Our model additionally accounts for non-saturating target concentrations and target dimerization, extending its application to more complex reactions. We used three novel beacon designs to validate the model, comparing single-and dual-target interactions and assessing temperature-dependent effects. Our approach captures both occupancy and temperature dependent complex behavior of multi-site and single-site beacons, revealing how a second binding event can enhance affinity and tune the dynamic range. Full derivations and design recommendations are provided, offering a predictive tool to guide the rational design of multi-site molecular beacons with controlled thermodynamics and fluorescent responses.
{"title":"Thermodynamic and Modeling Insights of DNA Molecular Beacons with Dual Target Binding to Design for Tunable Fluorescent Outputs","authors":"Esther Emelia Stopps, Stephanie Ellen McCalla","doi":"10.1039/d5an01208b","DOIUrl":"https://doi.org/10.1039/d5an01208b","url":null,"abstract":"Molecular beacons offer a versatile platform for detecting specific nucleic acid targets based on the relative proximity of fluorophore and quencher molecules. Conventional beacons contain a single target-binding site in the loop, and hybridization has been modeled based on three specific phases: closed hairpin, random coil, and target-bound. The target-beacon interactions can be tuned by stem length and target site location, while multi-site beacons introduce allosteric effects that modulate affinity and produce steeper, switch-like responses. Despite available thermodynamic models, current in silico methods do not adequately predict complex sensor outputs, especially for multi-site interactions. We present a new mathematical framework for calculating the thermodynamic parameters of beacons with two target binding sites. Our model additionally accounts for non-saturating target concentrations and target dimerization, extending its application to more complex reactions. We used three novel beacon designs to validate the model, comparing single-and dual-target interactions and assessing temperature-dependent effects. Our approach captures both occupancy and temperature dependent complex behavior of multi-site and single-site beacons, revealing how a second binding event can enhance affinity and tune the dynamic range. Full derivations and design recommendations are provided, offering a predictive tool to guide the rational design of multi-site molecular beacons with controlled thermodynamics and fluorescent responses.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"8 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146070163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiaodan Zhu, Guomin Gu, Yanli Shen, Mihray Abdurazik, Gang Sun
The precise and reliable identification of circular RNA (circRNA) is essential for both biological studies and clinical diagnostics of non-small cell lung cancer (NSCLC), especially the exosomal circRNA. In this study, we utilize the CRISPR/Cas13a system to specifically recognize the unique back-splice junction of target circRNA and develop a novel detection platform termed CRISPR/Cas13a-induced self-priming cyclic amplification. This method enables highly sensitive and specific circRNA detection. A pair of stem-loop DNA primers was carefully designed, each incorporating complementary single-stranded DNA sequences and five ribouridine (rU) residues at the 3′ end serving as an overhang. When Cas13a binds to the target circRNA, its trans-cleavage activity is activated, leading to the cleavage of the rU residues. This cleavage permits the 3’ ends of the stem-loop primers to extend along one another, generating multiple double stem-loop DNA structures that initiate successive cycles of self-priming chain elongation. By leveraging the sustained trans-cleavage activity of Cas13a and the high amplification efficiency of the self-priming cyclic reaction, the assay achieves sensitive detection of circRNA at concentrations as low as 564 aM within 90 min. In addition, the proposed method has been successfully applied for the analysis of exosomal hsa_circ_0003026 expression level in normal samples and NSCLC samples and demonstrated the potential of exosomal hsa_circ_0003026 in regulating the pathological progression. Owing to the high specificity of Cas13a, the proposed method can be directly applied to detect circRNA in complex biological samples without prior isolation of corresponding linear RNAs.
{"title":"CRISPR/Cas13a-Induced Self-Priming Cyclic Amplification Enables Liquid Biopsy of Exosomal Circular RNA in Non-small Cell Lung Cancer","authors":"Xiaodan Zhu, Guomin Gu, Yanli Shen, Mihray Abdurazik, Gang Sun","doi":"10.1039/d5an01345c","DOIUrl":"https://doi.org/10.1039/d5an01345c","url":null,"abstract":"The precise and reliable identification of circular RNA (circRNA) is essential for both biological studies and clinical diagnostics of non-small cell lung cancer (NSCLC), especially the exosomal circRNA. In this study, we utilize the CRISPR/Cas13a system to specifically recognize the unique back-splice junction of target circRNA and develop a novel detection platform termed CRISPR/Cas13a-induced self-priming cyclic amplification. This method enables highly sensitive and specific circRNA detection. A pair of stem-loop DNA primers was carefully designed, each incorporating complementary single-stranded DNA sequences and five ribouridine (rU) residues at the 3′ end serving as an overhang. When Cas13a binds to the target circRNA, its trans-cleavage activity is activated, leading to the cleavage of the rU residues. This cleavage permits the 3’ ends of the stem-loop primers to extend along one another, generating multiple double stem-loop DNA structures that initiate successive cycles of self-priming chain elongation. By leveraging the sustained trans-cleavage activity of Cas13a and the high amplification efficiency of the self-priming cyclic reaction, the assay achieves sensitive detection of circRNA at concentrations as low as 564 aM within 90 min. In addition, the proposed method has been successfully applied for the analysis of exosomal hsa_circ_0003026 expression level in normal samples and NSCLC samples and demonstrated the potential of exosomal hsa_circ_0003026 in regulating the pathological progression. Owing to the high specificity of Cas13a, the proposed method can be directly applied to detect circRNA in complex biological samples without prior isolation of corresponding linear RNAs.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"31 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146048795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mohamed W. Attwa, Haitham AlRabiah, Ali S. Abdelhameed, Adnan A. Kadi
Buparlisib (BLB) is an oral pan-phosphoinositide 3 kinase (PI3K) inhibitor that selectively targets all class I PI3K isoforms. No prior UPLC-MS/MS method with combined in silico metabolism analysis and green metrics for BLB has been reported, so this research sought to establish a precise, ultra-fast, sustainable, and dependable UPLC-MS/MS approach for estimating BLB in the matrix of human liver microsomes (HLMs), which is employed for determining the metabolic stability of BLB. The UPLC-MS/MS method exhibited a good degree of greenness as approved by the ComplexMoGAPI (69.0) and the AGREEprep tool score (0.68). Ripretinib (RPB) was selected as an internal standard (IS) for BLB quantification in the HLMs matrix. The validated method exhibited a linearity over an extensive concentration span of 1 to 4000 ng mL−1. The accuracy and precision of interday and intraday measurements varied from −2.92% and 10.11% and 3.11 and 9.78%, respectively. The MS/MS analysis was performed by employing the positive ESI ionization mode, and chromatography using an Eclipse Plus C8 column with a run time of one minute. BLB had a moderate extraction ratio, with a clearance rate (Clint) of 25.15 mL min−1 kg−1 and an in vitro half-life (t1/2) of 32.24 min. In silico assessments (P450 and DEREK modules) indicate that minor structural changes in the 2-aminopyridine moiety (lability: 58%) and morpholine groups (lability: 42%) during drug design could increase the metabolic stability and safety profile of BLB. The integrated in vitro technique (metabolic incubation) and in silico tools (ADME, DEREK and WhichP450) provide a resource-effective strategy for preliminary metabolic screening and advancing new therapeutic development aimed at enhancing metabolic stability of new BLB derivatives.
buparisib （BLB）是一种口服泛磷酸肌肽3激酶（PI3K）抑制剂，选择性靶向所有I类PI3K亚型。由于目前还没有将有机代谢分析与绿色指标相结合的UPLC-MS/MS方法报道，因此本研究试图建立一种精确、超快速、可持续、可靠的UPLC-MS/MS方法来估计人肝微粒体（HLMs）基质中的BLB，并用于确定BLB的代谢稳定性。通过ComplexMoGAPI（69.0）和AGREEprep工具评分（0.68），UPLC-MS/MS方法具有良好的绿色度。选择利普雷替尼（RPB）作为hms矩阵中BLB定量的内标（IS）。经过验证的方法在1至4000 ng mL−1的广泛浓度范围内呈现线性。日间和日间测量的准确度和精密度分别为- 2.92%和10.11%，3.11%和9.78%。MS/MS分析采用ESI正电离模式，色谱柱为Eclipse Plus C8，运行时间为1分钟。BLB提取率适中，清除率（Clint）为25.15 mL min−1 kg−1，体外半衰期（t1/2）为32.24 min。硅评估（P450和DEREK模块）表明，在药物设计过程中，2-氨基吡啶部分（不稳定性：58%）和morpholine基团（不稳定性：42%）的微小结构变化可以提高BLB的代谢稳定性和安全性。综合体外技术（代谢孵育）和计算机工具（ADME， DEREK和p450）为初步代谢筛选和推进新的治疗开发提供了一种资源有效的策略，旨在提高新的BLB衍生物的代谢稳定性。
{"title":"Quantification of buparlisib in human liver microsomes employing an ultra-fast, sensitive UPLC-MS/MS method: in vitro and in silico metabolic stability evaluation","authors":"Mohamed W. Attwa, Haitham AlRabiah, Ali S. Abdelhameed, Adnan A. Kadi","doi":"10.1039/d5an01155h","DOIUrl":"https://doi.org/10.1039/d5an01155h","url":null,"abstract":"Buparlisib (BLB) is an oral pan-phosphoinositide 3 kinase (PI3K) inhibitor that selectively targets all class I PI3K isoforms. No prior UPLC-MS/MS method with combined <em>in silico</em> metabolism analysis and green metrics for BLB has been reported, so this research sought to establish a precise, ultra-fast, sustainable, and dependable UPLC-MS/MS approach for estimating BLB in the matrix of human liver microsomes (HLMs), which is employed for determining the metabolic stability of BLB. The UPLC-MS/MS method exhibited a good degree of greenness as approved by the ComplexMoGAPI (69.0) and the AGREEprep tool score (0.68). Ripretinib (RPB) was selected as an internal standard (IS) for BLB quantification in the HLMs matrix. The validated method exhibited a linearity over an extensive concentration span of 1 to 4000 ng mL<small><sup>−1</sup></small>. The accuracy and precision of interday and intraday measurements varied from −2.92% and 10.11% and 3.11 and 9.78%, respectively. The MS/MS analysis was performed by employing the positive ESI ionization mode, and chromatography using an Eclipse Plus C8 column with a run time of one minute. BLB had a moderate extraction ratio, with a clearance rate (Cl<small><sub>int</sub></small>) of 25.15 mL min<small><sup>−1</sup></small> kg<small><sup>−1</sup></small> and an <em>in vitro</em> half-life (<em>t</em><small><sub>1/2</sub></small>) of 32.24 min. <em>In silico</em> assessments (P450 and DEREK modules) indicate that minor structural changes in the 2-aminopyridine moiety (lability: 58%) and morpholine groups (lability: 42%) during drug design could increase the metabolic stability and safety profile of BLB. The integrated <em>in vitro</em> technique (metabolic incubation) and <em>in silico</em> tools (ADME, DEREK and WhichP450) provide a resource-effective strategy for preliminary metabolic screening and advancing new therapeutic development aimed at enhancing metabolic stability of new BLB derivatives.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"274 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146048794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Human papillomavirus (HPV), particularly high-risk genotypes HPV16 and HPV18, is a leading cause of cervical cancer, contributing to significant global morbidity and mortality, especially in low-resource settings. Current diagnostic methods, such as PCR and cytology-based Pap tests, are limited by high costs, complex infrastructure requirements, and variable sensitivity, hindering their accessibility in low- and middle-income countries. To address these challenges, an integrated microfluidic platform was developed for rapid, sensitive, and point-of-care (POC) detection of HPV16 and HPV18 DNA. This platform combines loop-mediated isothermal amplification (LAMP) with lateral flow assay (LFA) detection, enabling a streamlined "sample-in-answer-out" workflow. The cartridge integrates nucleic acid (NA) extraction, isothermal amplification at 65 °C, and visual detection within 45 minutes, utilizing a lysis buffer optimized for sample preparation and genotype-specific primers targeting the E6 genes of HPV16 and HPV18. The microfluidic design leverages the Zweifach-Fung bifurcation principle for efficient NA separation, eliminating the need for centrifugation or external equipment. Analytical validation using spiked clinical samples demonstrated a limit of detection of 10 copies per µL, with T/C ratio-based quantification showing strong linearity (R2 > 0.98) over a dynamic range of 108 to 1 copy per µL. A clinical evaluation of 50 cervical swab samples yielded a sensitivity of 96.8% and a specificity of 100% compared to qPCR, with positive and negative predictive values of 100% and 94.7%, respectively. The platform's portability, low cost, and minimal user intervention make it an ideal solution for decentralized settings, offering a transformative approach to early HPV screening and cervical cancer prevention in resource-limited environments.
人乳头瘤病毒（HPV），特别是高危基因型HPV16和HPV18，是宫颈癌的主要病因，在全球造成重大发病率和死亡率，特别是在资源匮乏地区。目前的诊断方法，如聚合酶链反应和基于细胞学的巴氏试验，受到成本高、基础设施要求复杂和敏感性多变的限制，阻碍了低收入和中等收入国家获得这些方法。为了解决这些挑战，我们开发了一个集成的微流控平台，用于快速、敏感和即时检测HPV16和HPV18 DNA。该平台结合了环介导的等温扩增（LAMP）和横向流动分析（LFA）检测，实现了简化的“样品-样品-样品-输出”工作流程。该试剂盒集核酸（NA）提取、65°C等温扩增和45分钟内的目视检测于一体，利用针对HPV16和HPV18 E6基因的基因型特异性引物和优化的样品制备裂解缓冲液。微流控设计利用茨威法赫-冯分岔原理进行高效NA分离，无需离心或外部设备。使用加标临床样品的分析验证表明，检测限为每μ L 10个拷贝，基于T/C比的定量在每μ L 108至1个拷贝的动态范围内显示出很强的线性（R2 > 0.98）。与qPCR相比，对50份宫颈拭子样本的临床评估结果显示，该方法的敏感性为96.8%，特异性为100%，阳性预测值为100%，阴性预测值为94.7%。该平台的便携性、低成本和最少的用户干预使其成为分散环境的理想解决方案，在资源有限的环境中为早期HPV筛查和宫颈癌预防提供了一种变革性方法。
{"title":"Integrated microfluidic device for rapid and accurate point-of-care detection of high-risk HPV16 and HPV18.","authors":"Natish Kumar, Monika Kumari, Devtulya Chander, Sandeep Dogra, Asha Chaubey, Ravi Kumar Arun","doi":"10.1039/d5an01244a","DOIUrl":"https://doi.org/10.1039/d5an01244a","url":null,"abstract":"<p><p>Human papillomavirus (HPV), particularly high-risk genotypes HPV16 and HPV18, is a leading cause of cervical cancer, contributing to significant global morbidity and mortality, especially in low-resource settings. Current diagnostic methods, such as PCR and cytology-based Pap tests, are limited by high costs, complex infrastructure requirements, and variable sensitivity, hindering their accessibility in low- and middle-income countries. To address these challenges, an integrated microfluidic platform was developed for rapid, sensitive, and point-of-care (POC) detection of HPV16 and HPV18 DNA. This platform combines loop-mediated isothermal amplification (LAMP) with lateral flow assay (LFA) detection, enabling a streamlined \"sample-in-answer-out\" workflow. The cartridge integrates nucleic acid (NA) extraction, isothermal amplification at 65 °C, and visual detection within 45 minutes, utilizing a lysis buffer optimized for sample preparation and genotype-specific primers targeting the E6 genes of HPV16 and HPV18. The microfluidic design leverages the Zweifach-Fung bifurcation principle for efficient NA separation, eliminating the need for centrifugation or external equipment. Analytical validation using spiked clinical samples demonstrated a limit of detection of 10 copies per µL, with T/C ratio-based quantification showing strong linearity (<i>R</i><sup>2</sup> > 0.98) over a dynamic range of 10<sup>8</sup> to 1 copy per µL. A clinical evaluation of 50 cervical swab samples yielded a sensitivity of 96.8% and a specificity of 100% compared to qPCR, with positive and negative predictive values of 100% and 94.7%, respectively. The platform's portability, low cost, and minimal user intervention make it an ideal solution for decentralized settings, offering a transformative approach to early HPV screening and cervical cancer prevention in resource-limited environments.</p>","PeriodicalId":63,"journal":{"name":"Analyst","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146049691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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