Analytical Chemistry最新文献

{"title":"Interfacial Charge Regulation by Phosphate Adsorption on Indium Tin Oxide for Charge-Selective Electrochemical Sensing","authors":"Yuri Kim,&nbsp;Jeongse Yun,&nbsp;Yerin Bang,&nbsp;Jungwon Kim,&nbsp;Seol Baek,&nbsp;Seung-Ryong Kwon,&nbsp;Minjee Seo","doi":"10.1021/acs.analchem.6c02827","DOIUrl":"10.1021/acs.analchem.6c02827","url":null,"abstract":"<p>Controlling interfacial charge withoutcompromising mass transportremains a challenge in charge-selective electrochemical sensing. Here,we use phosphate adsorption on indium tin oxide (ITO) as a model systemto investigate how specifically adsorbed anions regulate the apparentelectron-transfer kinetics and enable charge-selective electrochemicalsensing. Using three ferrocene derivatives with distinct charge states,we show that phosphate adsorption enhances the voltammetric responseof cationic species while suppressing that of anionic species, ina manner strongly dependent on pH and phosphate speciation. To elucidatethe underlying mechanism, we directly quantify shifts in the potentialof zero charge using scanning electrochemical cell microscopy (SECCM)and correlate these measurements with the observed kinetic trends.Comparison of orthophosphate, pyrophosphate, and tripolyphosphatedemonstrates that multidentate adsorption and higher anion chargeproduce progressively stronger electrostatic modulation. Leveragingthis effect, we achieve charge-selective electro-oxidation of dopaminein the presence of ascorbic acid and uric acid, with oligophosphateadsorption further enhancing the discrimination of dopamine and yieldingdetection limits of 0.15 and 0.11 μM for pyrophosphate and tripolyphosphate,respectively. The reversibility of phosphate adsorption under mildalkaline treatment further establishes this approach as a simple andregenerable alternative to conventional film-based interface engineering.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29493–29506"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149049910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alternative to Nanoluminophores: Multiresonant 1,4-BN Emitters for Waveband- and Monochromaticity-Tunable Electrochemiluminescence","authors":"Yujiao Wang,&nbsp;Wendong Jin,&nbsp;Shengrong Chen,&nbsp;Shengjie Li,&nbsp;Yue Sun,&nbsp;Zhiqiang Liu,&nbsp;Guizheng Zou","doi":"10.1021/acs.analchem.6c04830","DOIUrl":"10.1021/acs.analchem.6c04830","url":null,"abstract":"<p>Nanoluminophoresare promising candidates for monochromaticemission;however, the commercial and quantitative applications of nanoluminophoresare hindered by their undefined molar mass, limited storage stability,and lack of reproducibility in fabrication. Here, we propose a 1,4-boron–nitrogen(1,4-BN)-embedded multiresonance structure (CzBNs) to design molecularluminophores with electrochemiluminescence (ECL) emission tunablein waveband and monochromaticity, which is comparable to the ECL luminophoresof deliberately designed nanomaterials and superior to the commercialECL luminophores based on conventional molecules. Moreover, the molecularluminophore CzBN-Ph–COOH can be utilized as an ECL tag in asandwich-type immunosensor to accurately determine carcinoembryonicantigen (CEA) at the 10 pg/mL level without the inherent limitationsof nanomaterials.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29676–29683"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149091609","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Subpicogram-Level Quantification of Trace Biomarkers Using an Automated High-Sensitivity Chemiluminescence Immunoassay Technology Based on a Novel Substrate Enhancement Strategy","authors":"Kexin Li,&nbsp;Zhuonan Duan,&nbsp;Ke Li,&nbsp;Yanzhong Wang,&nbsp;Minjie Wang,&nbsp;Yujuan Zhang,&nbsp;Ruijun Tian,&nbsp;Changying Fu,&nbsp;Yushu Yuan,&nbsp;Chaoman Ang,&nbsp;Xueqing Liu,&nbsp;Hui Ma,&nbsp;Jun Zhang,&nbsp;Xin Zhang,&nbsp;Wei Cui","doi":"10.1021/acs.analchem.6c03190","DOIUrl":"https://doi.org/10.1021/acs.analchem.6c03190","url":null,"abstract":"<p>Trace biomarkers showed great potential for improvingthe diagnosisand monitoring of various diseases. However, the sensitivity of thecurrent high-throughput detection platforms is limited, posing a significantchallenge for the reliable measurement and clinical application oftrace biomarkers. We developed an automated high-sensitivity chemiluminescenceimmunoassay (CLIA) technology with subpicogram-level sensitivity basedon the modification of the substrate molecular structure and the synergyof surfactant and fluorescein. Using this technology, we constructedassay kits for five trace biomarkers: leukemia inhibitory factor (LIF),high-sensitivity cardiac troponin I (hs-cTnI) and T (hs-cTnT), phosphorylatedtau-217 (p-tau217), and estradiol (E2). The analytical and clinicalperformance was evaluated by examining the precision, linearity, limitsof detection, and methodological comparison. The high-sensitivitychemiluminescent substrate demonstrated approximately 7-fold highersensitivity and 2.5-fold greater luminescence efficiency comparedto those of the traditional substrate, with a linear range exceeding160 million relative light units (RLUs). The five trace biomarkerassays exhibited excellent analytical sensitivity, with the LoD valuesof 0.070 pg/mL (LIF), 0.18 pg/mL (hs-cTnI), 1.1 pg/mL (hs-cTnT), 0.007pg/mL (p-tau217), and 0.113 pg/mL (E2 II). Compared with other high-sensitivitydetection methods, the Passing–Bablok linear correlation coefficients(<em>r</em>) ranged from 0.9767 to 0.9974. The proportionsof cTnI, cTnT, and E2 II concentrations above the LoD in healthy populationsexceeded 98%. We established an automated high-sensitivity CLIA technologybased on a novel substrate enhancement strategy, improving the analyticalsensitivity to the subpicogram level. An automated high-throughputdetection method of various trace biomarkers was realized that mayhave broad application in the clinic.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29553–29564"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149218983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Demystifying Conformational Preorganization in Pyrene-Imidazopyridine Probes: Complementary Roles of π-Spacer Integration and Zn2+ Templating in Sequential Cu2+ Recognition","authors":"Bhuvaneesh Ilango,Arunkumar Kathiravan,Marappan Velusamy","doi":"10.1021/acs.analchem.6c06203","DOIUrl":"https://doi.org/10.1021/acs.analchem.6c06203","url":null,"abstract":"The analytical performance of fluorescent chemosensors is fundamentally governed by the interplay among intramolecular charge transfer efficiency, coordination-site accessibility, and receptor−analyte binding affinity. However, in compact fluorophore−receptor scaffolds, planar fluorophores and sterically demanding receptor units can impose torsional strain, attenuating donor−acceptor electronic transitions while restricting analyte access to the coordination cavity. To delineate this structure−performance relationship, a sterically congested D−A probe was investigated as a structural counterpart to its phenyl π-spacer-integrated D−π−A analogue. The π-spacer serves as an internal preorganization unit that relieves steric strain, preserves near-global coplanarity, improves receptor accessibility, and promotes ICT characteristics. As a result, the D−π−A probe exhibited stronger affinity toward Cu2+ and Zn2+ ions with superior detection limits of 1.86 nM and 0.85 nM, respectively. Complementing the spacer-mediated internal preorganization, Zn2+ coordination provides an external supramolecular template that organizes the receptor into a binding-competent geometry for sequential Cu2+ recognition. This template effect was more pronounced for the sterically congested D−A probe, increasing its apparent Cu2+ binding constant by nearly 1 order of magnitude (1.59 × 105 M−1) with an improved detection limit of 1.06 nM. Nevertheless, the D−π−A + Zn2+ complex remained the most sensitive system, achieving an enhanced detection limit of 0.72 nM. Furthermore, the practical applicability of the probes was demonstrated by their reliable performance in mixed aqueous media, sensitive recognition of Cu-GHK, spike-recovery studies, and robust performance in both fluorescent paper-strip assays and live-cell imaging across mammalian and plant cells. Overall, this work establishes π-spacer-mediated internal preorganization and Zn2+ template-assisted external preorganization as complementary strategies for designing efficient fluorescent chemosensors.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"12 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multivariate Metalloporphyrin-Based Covalent Organic Frameworks for Cataluminescence Signal Amplification","authors":"Ruidong Chen,&nbsp;Jincai Lu,&nbsp;Yaze Chen,&nbsp;Yuhao Liu,&nbsp;Chao Lu","doi":"10.1021/acs.analchem.6c04360","DOIUrl":"10.1021/acs.analchem.6c04360","url":null,"abstract":"<p>Utilizingcovalent organic frameworks (COFs) without postpyrolysisfor cataluminescence (CTL) remains a challenge due to deficient thermocatalyticactivity and uncontrollable bimetallic ratios. Herein, we presenteda multivariate (MTV) metalloporphyrin-based COF (Cu_0.5/Ni_0.5-COF) embedded with nonpyrolysis and atomically dispersedbimetallic sites as a high-performance CTL platform. Interestingly,the Cu_0.5/Ni_0.5-COF catalyst exhibited an extraordinarysignal amplification of diethyl ether-based CTL as a result of themaximum exposure of the bimetallic active centers and the enrichmentof Lewis acid sites. The developed CTL sensor delivered a state-of-the-artsensing profile, including an ultrafast response time of 0.8 s, awide linear range (0.1–15.0 mM), and a remarkable detectionlimit of 0.02 mM. The excellent performances of the as-prepared Cu_0.5/Ni_0.5-COF catalyst were ascribed to the profoundsynergistic interaction between the bimetallic porphyrin centers andthe analyte for high-efficiency light emissions. This work opens anew avenue for engineering pristine COF-based CTL platforms.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29648–29656"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149092677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioinspired Compartmentalization Coupled with DNA-Gated Magnetic Nanoprobes Enables Simultaneous Electrochemical Detection of Dual MicroRNAs","authors":"Jiangbo Dong,Beibei Yu,Lili Zou,Ke Lv,Guoming Xie,Gang Yang","doi":"10.1021/acs.analchem.6c04958","DOIUrl":"https://doi.org/10.1021/acs.analchem.6c04958","url":null,"abstract":"Simultaneous analysis of multiple microRNAs (miRNAs) is highly desirable for reliable molecular diagnosis, yet multiplexed amplification assays are often compromised by nonspecific amplification, signal crosstalk, and background leakage. Inspired by biological compartmentalization, we developed a spatially isolated and DNA-gated electrochemical sensing platform for the simultaneous detection of miRNA-21 and miRNA-155. A custom dual-chamber polymerase chain reaction (PCR) tube was employed to physically separate two target-triggered exponential amplification reactions, allowing each miRNA to generate its corresponding single-stranded DNA product independently. The amplified products subsequently acted as molecular keys to open sequence-programmed DNA gates on mesoporous Fe3O4 magnetic nanoprobes loaded with electroactive reporters. Through toehold-mediated strand displacement, miRNA-21-derived products triggered ferrocene release, while miRNA-155-derived products induced methylene blue release, producing two distinguishable differential pulse voltammetry signals on screen-printed electrodes. By integrating spatially orthogonal amplification with programmable gated release, the proposed strategy effectively suppressed inter-reaction interference and converted trace miRNA inputs into amplified electrochemical outputs. Under optimized conditions, the sensor exhibited a wide linear range from 1 fM to 10 nM, with detection limits of 0.96 fM for miRNA-21 and 0.83 fM for miRNA-155. The platform also showed excellent selectivity, satisfactory reproducibility, good storage stability, and reliable recoveries in diluted human serum. Furthermore, it successfully differentiated MCF-7 breast cancer cells from LO2 control cells, with expression trends consistent with reverse transcription quantitative PCR (RT-qPCR). This compartmentalized DNA-gated sensing strategy provides a versatile and interference-resistant route for multiplexed nucleic acid analysis and holds promise for early cancer-related molecular diagnostics.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"71 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149221715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Task-Dependent Synergy between FTIR and Raman Spectroscopy for Sperm Quality Prediction","authors":"Beatrice Campanella,&nbsp;Lorenzo Birindelli,&nbsp;Stefano Legnaioli,&nbsp;Giulia Lorenzetti,&nbsp;Cristina Consani,&nbsp;Valeria Ales,&nbsp;Edoardo Benedetti,&nbsp;Maria Antonella Bertozzi,&nbsp;Emilia Bramanti","doi":"10.1021/acs.analchem.6c04960","DOIUrl":"10.1021/acs.analchem.6c04960","url":null,"abstract":"<p>Maleinfertility affects approximately 15% of couples worldwide,contributing to 40–50% of infertility cases in Western countries.Standard semen analysis remains the cornerstone of fertility assessment,yet conventional parameters (concentration, motility, morphology)have limited predictive value for fertilization success. Vibrationalspectroscopy, particularly Fourier transform infrared (FTIR) and Ramanspectroscopy, offers rapid, nondestructive chemical fingerprintingof biological samples. However, optimal strategies for integratingmultimodal spectroscopic data remain unclear, and the value of fusionmay depend on the biological end point. This study systematicallycompares FTIR and Raman spectroscopy, both independently and throughmultiple fusion strategies, for predicting rectilinear progressivesperm motility (binary classification) and normal morphology (continuousregression) in human seminal plasma. We determine whether complementaryinformation exists, when fusion provides value, and which strategyoptimally leverages multimodal data. Building on Raman-derived MultivariateCurve Resolution-Alternating Least Squares (MCR-ALS) concentrationprofiles, we further develop a clinical classifier based on four seminalplasma biomarkers (human serum albumin, glutathione S-transferase,transferrin, lysophosphatidylcholine), validated by leave-one-patient-outcross-validation and Firth’s penalized logistic regressionto address near-complete separation in this small-sample setting (n= 30). Our findings reveal technique-specific advantages: Raman spectroscopysubstantially outperforms FTIR for binary motility classification,with feature-level fusion matching but not exceeding Raman’sperformance, indicating limited complementarity. Conversely, FTIRalone achieves superior predictive accuracy for continuous morphologicaloutcomes. The clinical classifier based on MCR-ALS-derived biomarkersachieved AUC (Area Under the ROC Curve) = 0.855, accuracy = 86.7%,and BER (Balanced Error Rate) = 0.136, with model significance confirmedby permutation testing. Elevated human serum albumin and lysophosphatidylcholineemerged as the strongest negative predictors of high motility, consistentwith seminal tract inflammation and sperm-cytotoxic phospholipid accumulation,while transferrin showed a positive association in line with its rolein iron availability for sperm function. These results demonstratethat optimal spectroscopic approaches depend on the biological endpoint, with Raman’s structural sensitivity favoring motilityassessment and FTIR’s compositional profiling better capturingmorphological variation. The biomarker-based classifier is easierto interpret and complements whole-spectrum models by bridging chemometricanalysis and biological interpretation.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29684–29699"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148973812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"p-Type Dopant-Mediated Electron–Hole Recombineering to Tune the Electrochemiluminescence of Metal–Organic Frameworks","authors":"Guijun Li,&nbsp;Shuai Luo,&nbsp;Si Zhang,&nbsp;Mengjiao Li,&nbsp;Huipu Liu,&nbsp;Huangxian Ju","doi":"10.1021/acs.analchem.6c03746","DOIUrl":"10.1021/acs.analchem.6c03746","url":null,"abstract":"<p>Constructing high-performance electrochemiluminescence(ECL) emittersthrough rational structural engineering is pivotal to overcoming theintrinsic limitations of conventional systems and advancing theirpractical applications. This work reports an electron–holerecombination engineering strategy to enhance the ECL emission ofmetal–organic frameworks (MOFs) by p-type doping of metal single-atomsites. Using Zr-TBAPy as a model, the experimental data and theoreticalcalculations demonstrate that the doping of single-atom Au inducesnotable defect energy levels, thereby providing additional recombinationchannels for conduction-band (CB) electrons and valence-band (VB)holes. The defect level ultimately increases the electron–holerecombination efficiency. Compared to Zr-TBAPy, single-atom Au-dopedZr-TBAPy (Au@Zr-TBAPy) exhibits a 4-fold enhancement in cathodic ECLintensity with an ECL efficiency of up to 50.56%. An ECL immunoassaywas used to demonstrate the ECL application of Au@Zr-TBAPy. This worknot only proposes a rational metal single-atom doping strategy forengineering ECL emitters to achieve high performance but also providesfundamental insights into the dependence of ECL emission on metalsingle-atom sites, opening new avenues for advanced applications ofMOFs in sensitive ECL biosensing.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29579–29587"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149019588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Real-Time Monitoring Ferroptosis-Mediated Drug-Induced Liver Injury Using a Near-Infrared Mitochondrial Viscosity Probe","authors":"Ben Liu,&nbsp;Yiwu Qiu,&nbsp;Long He,&nbsp;Yi Li","doi":"10.1021/acs.analchem.6c04129","DOIUrl":"10.1021/acs.analchem.6c04129","url":null,"abstract":"<p>Drug-induced liver injury (DILI) is a frequent clinicalconditionin which hepatocyte ferroptosis serves as a core pathological mechanism,leading to critical alterations in the mitochondrial microenvironment,including viscosity. However, visualizing mitochondrial viscositychanges in real time during ferroptosis-driven DILI remains technicallychallenging. Herein, we report XT-TPE, a mitochondrion-targeted near-infrared(NIR) “turn-on” viscosity probe based on a tetraphenylethylene(TPE)-xanthene hybrid. This probe gives a remarkable 129-fold fluorescenceincrease in high-viscosity media with an outstanding linear relationship(<em>R</em>² &gt; 0.999). The probe displays lowcytotoxicitytoward hepatocytes, excellent mitochondrial colocalization (Pearson’scoefficient: 0.916), and enables sensitive monitoring of viscosityfluctuations during ferroptosis-mediated DILI both <em>in vitro</em> and <em>in vivo</em>. Using this probe, we screened 14 redox-activecompounds and identified N-acetylcysteine (NAC) and ferrostatin-1(Fer-1) as strong inhibitors of ferroptosis-driven DILI. In acetaminophen-inducedDILI mouse models, XT-TPE enables noninvasive hepatic viscosity imagingand confirms the therapeutic efficacy of NAC and Fer-1. This workprovides a reliable molecular tool for ferroptosis-related viscosityimaging and establishes a new strategy for anti-DILI drug screening.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29616–29626"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149049986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantification of Intact MDa Poly(A)-Tailed and Non-Tailed mRNA Fractions with Charge Detection Mass Spectrometry","authors":"Julian Robles,&nbsp;Zachary M. Miller,&nbsp;Aaron M. Roe,&nbsp;Stephanie M. Thibert,&nbsp;Matthew R. Gardner,&nbsp;Janelle Tricia M. Magaling,&nbsp;Justin W. Torpey,&nbsp;Angelica F. Castañeda,&nbsp;Evan R. Williams","doi":"10.1021/acs.analchem.6c03007","DOIUrl":"10.1021/acs.analchem.6c03007","url":null,"abstract":"<p>The clinical success of messenger ribonucleic acid (mRNA)therapeuticshas driven the need for robust analytical techniques capable of characterizingmixtures of megadalton (MDa) constructs. A critical attribute of thesetherapeutics is the presence and length of the 3′ poly(A) tail,which governs mRNA stability and translational efficiency. Chargedetection mass spectrometry (CDMS) was used to distinguish and quantifyintact MDa mRNA variants differing only by the poly(A) content. Initialanalysis under native-like conditions was limited by salt adduction,but a 75% methanol solution resulted in nearly a five-fold increasein charge (from ∼60<em>e</em> to ∼280<em>e</em>) and substantially less adduction. Measured masses ofthe intact mRNA were 1.054 MDa and 1.027 MDa for the tailed and non-tailedmRNA constructs, respectively, consistent with the theoretical 26.6kDa mass difference. Mixtures of various compositions resulted inbroad, unresolved peaks at intermediate mass owing to residual saltadduction. A constrained Gaussian deconvolution of these data enabledaccurate quantification across seven different mole fractions (<em>R</em>² = 0.978). Notably, an analysis of a subpopulationof the highest charge-state ions revealed separation into two distinctpeaks, demonstrating that mass resolution for MDa mRNA analysis islimited by adduction rather than instrumental factors, motivatingadditional studies on methods to increase charging for more accuratequantification. These results establish CDMS as a powerful, top-downmethod for precise monitoring of mRNA heterogeneity and degradationin high molecular weight therapeutics.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"98 41","pages":"29507–29514"},"PeriodicalIF":7.3,"publicationDate":"2026-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149091910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}