Vibrational spectroscopy has gained significant attention in medical diagnosis due to its high sensitivity and nondestructive nature. Raman spectroscopy and infrared spectroscopy complement each other in their selection rules, vibration responses, and wavenumber coverage. Combining these two techniques can overcome the limitations of individual spectra, enhancing the accuracy of molecular structure identification. However, existing deep learning fusion methods often overlook the diagnostic advantages of different modalities, leading to overreliance on strong modalities or interference from weak modality noise, resulting in unstable fusion and imbalanced information flow. We propose a Symbiotic Attention Fusion Decoupled Network (SAFDN) to effectively model the information guidance mechanism. In the prefusion stage, we combine multilayer perceptrons and convolutional neural networks for intramodal encoding, laying the foundation for cross-modal fusion. Then, we design Symbiotic Attention Fusion (SAF) and Parasitic Attention Fusion (PAF) mechanisms to simulate biological symbiosis and parasitism, achieving a differentiated information enhancement. Finally, a supervised multimodal contrastive learning decoupling network is introduced to balance cross-modal consistency and intramodal cohesion, improving feature decoupling and semantic fusion. Experiments on cancer, autoimmune diseases, and cardiovascular disease data sets show that SAFDN outperforms existing methods, achieving accuracy and AUC values of 90.49%/0.9649, 95.48%/0.9866, and 96.67%/0.9934, respectively. SAFDN validates the advantages of the symbiotic effect in vibrational spectroscopy disease classification tasks through an in-depth comparison and analysis of fusion and loss mode ratios. This model provides an efficient solution for rapid, noninvasive precision medical diagnosis, improving the accuracy and interpretability of disease classification.
{"title":"Information-Guided Fusion of Multimodal Vibrational Spectroscopy for Disease Diagnosis Based on Symbiotic Attention Decoupled Contrastive Learning","authors":"Xuguang Zhou,Wenjie Fan,Chen Chen,Xiangnan Chen,Yining Yang,Lijun Wu,Jin Gu,Lei Yan,Jing Tao,Xue Wu,Xiaoyi Lv,Cheng Chen","doi":"10.1021/acs.analchem.5c06086","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c06086","url":null,"abstract":"Vibrational spectroscopy has gained significant attention in medical diagnosis due to its high sensitivity and nondestructive nature. Raman spectroscopy and infrared spectroscopy complement each other in their selection rules, vibration responses, and wavenumber coverage. Combining these two techniques can overcome the limitations of individual spectra, enhancing the accuracy of molecular structure identification. However, existing deep learning fusion methods often overlook the diagnostic advantages of different modalities, leading to overreliance on strong modalities or interference from weak modality noise, resulting in unstable fusion and imbalanced information flow. We propose a Symbiotic Attention Fusion Decoupled Network (SAFDN) to effectively model the information guidance mechanism. In the prefusion stage, we combine multilayer perceptrons and convolutional neural networks for intramodal encoding, laying the foundation for cross-modal fusion. Then, we design Symbiotic Attention Fusion (SAF) and Parasitic Attention Fusion (PAF) mechanisms to simulate biological symbiosis and parasitism, achieving a differentiated information enhancement. Finally, a supervised multimodal contrastive learning decoupling network is introduced to balance cross-modal consistency and intramodal cohesion, improving feature decoupling and semantic fusion. Experiments on cancer, autoimmune diseases, and cardiovascular disease data sets show that SAFDN outperforms existing methods, achieving accuracy and AUC values of 90.49%/0.9649, 95.48%/0.9866, and 96.67%/0.9934, respectively. SAFDN validates the advantages of the symbiotic effect in vibrational spectroscopy disease classification tasks through an in-depth comparison and analysis of fusion and loss mode ratios. This model provides an efficient solution for rapid, noninvasive precision medical diagnosis, improving the accuracy and interpretability of disease classification.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"314 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139077","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}
Secondary-ion mass spectrometry (SIMS) and related in situ microanalytical techniques allow precise chemical and isotopic characterization at micron and submicron scales, providing insights into spatially heterogeneous processes. However, the quantitative accuracy of SIMS is limited by matrix effects, which cause instrumental mass fractionation (IMF) between measured and true isotope ratios. Accurate quantification requires matrix-matched reference materials (RMs) with identical physical and chemical properties to the unknown samples. Most existing SIMS RMs are derived from natural minerals, which often exhibit heterogeneity, limiting reproducibility and interlaboratory comparability. Synthetic RMs offer a promising solution, but their development for SIMS has been challenging due to the sensitivity of SIMS to microstructural attributes such as surface smoothness and grain size. This study presents a novel synthesis strategy for producing matrix-matched pyrite (FeS2) RMs. By combining hydrothermal precursor synthesis with low-temperature ultrahigh-pressure (UHP) sintering, we fabricated dense, nanocrystalline pyrite ceramics with controlled stoichiometry and exceptional sulfur-isotope homogeneity. The resulting material exhibited sputtering behavior indistinguishable from that of natural pyrite, demonstrating the strategy as a robust framework for producing synthetic sulfide RMs. This approach facilitates the improvement of analytical accuracy and reproducibility in microanalytical science and can be extended to other mineral systems.
{"title":"A Novel Strategy for Synthesizing Matrix-Matched Microanalysis Reference Materials via Hydrothermal Precursors and Ultrahigh-Pressure Sintering","authors":"Youwei Chen,Sen Lin,Jian-Feng Gao,Xianwu Bi,Guoqiang Tang,Zhian Bao,Qing Yang,Zexian Cui,Shaohua Dong,Ruizhong Hu","doi":"10.1021/acs.analchem.5c07701","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07701","url":null,"abstract":"Secondary-ion mass spectrometry (SIMS) and related in situ microanalytical techniques allow precise chemical and isotopic characterization at micron and submicron scales, providing insights into spatially heterogeneous processes. However, the quantitative accuracy of SIMS is limited by matrix effects, which cause instrumental mass fractionation (IMF) between measured and true isotope ratios. Accurate quantification requires matrix-matched reference materials (RMs) with identical physical and chemical properties to the unknown samples. Most existing SIMS RMs are derived from natural minerals, which often exhibit heterogeneity, limiting reproducibility and interlaboratory comparability. Synthetic RMs offer a promising solution, but their development for SIMS has been challenging due to the sensitivity of SIMS to microstructural attributes such as surface smoothness and grain size. This study presents a novel synthesis strategy for producing matrix-matched pyrite (FeS2) RMs. By combining hydrothermal precursor synthesis with low-temperature ultrahigh-pressure (UHP) sintering, we fabricated dense, nanocrystalline pyrite ceramics with controlled stoichiometry and exceptional sulfur-isotope homogeneity. The resulting material exhibited sputtering behavior indistinguishable from that of natural pyrite, demonstrating the strategy as a robust framework for producing synthetic sulfide RMs. This approach facilitates the improvement of analytical accuracy and reproducibility in microanalytical science and can be extended to other mineral systems.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"39 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139081","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}
Pub Date : 2026-02-09DOI: 10.1021/acs.analchem.5c08055
Wenjing Liu,Wei Chen,Wei Li,Minghao Liu,Pengfei Tu,Haiyu Zhao,Yuelin Song
Although derivatization is widely favored for metabolomics, the applications are dramatically narrowed by insufficient selectivity, because a given metabolite may be transferred to several products or conjugated with two or even more derivative moieties. Enzymatic derivatization may address this critical issue by attributing to superior selectivity. Human sulfotransferase 2A1 (hSULT 2A1) was utilized here to tag sulfo to 3-OH of steroids that structurally involve most cholesterol metabolites and, importantly, serve as key biomarkers for diverse diseases. Through evaluating sulfation performances by assaying 53 authentic steroids, we found: 1) great selectivity and transformation rate (>80%) existed for 3-OH sulfation; 2) sulfates exhibited diagnostic fragment ions (i.e., SO3–• and SO4–), SO3 neutral loss, and [34S – M – H]− signals; 3) optimal collision energy for either SO3–• or SO4– was linearly correlated with [M – H]− mass; and 4) better sensitivity appeared for sulfates. Molecular docking consolidated selective 3-OH sulfation. hSULT 2A1-catalyzed sulfo-tagging was applied for 3-OH steroid-targeted submetabolome profiling of Bufonis Venenum (BV), a promising anticancer agent. Sixty steroid 3-sulfates were captured and quantitatively compared, and significant variations existed within 20 batches of BV. Together, hSULT 2A1-mediated sulfation is meaningful for submetabolomics targeting on 3-OH steroids, leading to new insights toward enzyme-catalyzed derivative metabolomics.
{"title":"Enzyme-Catalyzed Sulfo-Tagging Accelerates 3-OH Steroid-Targeted Submetabolome Profiling","authors":"Wenjing Liu,Wei Chen,Wei Li,Minghao Liu,Pengfei Tu,Haiyu Zhao,Yuelin Song","doi":"10.1021/acs.analchem.5c08055","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c08055","url":null,"abstract":"Although derivatization is widely favored for metabolomics, the applications are dramatically narrowed by insufficient selectivity, because a given metabolite may be transferred to several products or conjugated with two or even more derivative moieties. Enzymatic derivatization may address this critical issue by attributing to superior selectivity. Human sulfotransferase 2A1 (hSULT 2A1) was utilized here to tag sulfo to 3-OH of steroids that structurally involve most cholesterol metabolites and, importantly, serve as key biomarkers for diverse diseases. Through evaluating sulfation performances by assaying 53 authentic steroids, we found: 1) great selectivity and transformation rate (>80%) existed for 3-OH sulfation; 2) sulfates exhibited diagnostic fragment ions (i.e., SO3–• and SO4–), SO3 neutral loss, and [34S – M – H]− signals; 3) optimal collision energy for either SO3–• or SO4– was linearly correlated with [M – H]− mass; and 4) better sensitivity appeared for sulfates. Molecular docking consolidated selective 3-OH sulfation. hSULT 2A1-catalyzed sulfo-tagging was applied for 3-OH steroid-targeted submetabolome profiling of Bufonis Venenum (BV), a promising anticancer agent. Sixty steroid 3-sulfates were captured and quantitatively compared, and significant variations existed within 20 batches of BV. Together, hSULT 2A1-mediated sulfation is meaningful for submetabolomics targeting on 3-OH steroids, leading to new insights toward enzyme-catalyzed derivative metabolomics.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"45 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139048","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}
The detection of related substances in both active pharmaceutical ingredients (APIs) and dosage forms, especially via process analytical technology (PAT), is of crucial importance for the assurance of pharmaceutical quality and clinical safety. However, this process remains a formidable challenge because related substances are structurally similar to APIs, resulting in severe spectral overlap. Herein, we present a host–guest doping room-temperature phosphorescence (RTP) sensing strategy that facilitates rapid detection of impurities in naproxen APIs and dosage forms. Leveraging the low luminescence efficiency at minimal host content, the platform can sensitively detect trace impurities, 2-acetyl-6-methoxynaphthalene (MANAP), in naproxen APIs, achieving a limit of detection (LOD) of 0.05% (w/w), which satisfies the pharmacopeial threshold of 0.1% (w/w). The method demonstrates statistical equivalency to HPLC, with average recovery rates of 98.03%–103.38%. Furthermore, both spectral analysis and real-time visualization inspection were successfully achieved for the limit test of MANAP in naproxen granules and tablets. This work introduces a novel RTP-based PAT approach for impurity testing in pharmaceutical manufacturing.
{"title":"Host–Guest Doping Room-Temperature Phosphorescence Sensing Strategy for the Detection of Related Substances in Naproxen APls and in the Production of Dosage Forms","authors":"Xiwen Zhu,Shishi Shao,Jialu Zheng,Xuhui Ma,Yan Jie Li,Jing Gao,Guo-Gang Shan,Cheng Zhi Huang,Peng Fei Gao","doi":"10.1021/acs.analchem.5c07536","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07536","url":null,"abstract":"The detection of related substances in both active pharmaceutical ingredients (APIs) and dosage forms, especially via process analytical technology (PAT), is of crucial importance for the assurance of pharmaceutical quality and clinical safety. However, this process remains a formidable challenge because related substances are structurally similar to APIs, resulting in severe spectral overlap. Herein, we present a host–guest doping room-temperature phosphorescence (RTP) sensing strategy that facilitates rapid detection of impurities in naproxen APIs and dosage forms. Leveraging the low luminescence efficiency at minimal host content, the platform can sensitively detect trace impurities, 2-acetyl-6-methoxynaphthalene (MANAP), in naproxen APIs, achieving a limit of detection (LOD) of 0.05% (w/w), which satisfies the pharmacopeial threshold of 0.1% (w/w). The method demonstrates statistical equivalency to HPLC, with average recovery rates of 98.03%–103.38%. Furthermore, both spectral analysis and real-time visualization inspection were successfully achieved for the limit test of MANAP in naproxen granules and tablets. This work introduces a novel RTP-based PAT approach for impurity testing in pharmaceutical manufacturing.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"70 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139080","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}
Pub Date : 2026-02-09DOI: 10.1021/acs.analchem.5c04490
Ding-Gui Cai,Shufen Pan,Teng-Fei Zheng,Chen Cao,Zhao-Bo Hu,Yan Peng,Yongquan Wu,He-Rui Wen,Sui-Jun Liu
The rapid detection of specific biological substances and biomarkers has the potential to contribute to the early identification, diagnosis, and prevention of diseases. Two examples of structurally similar luminescent metal–organic frameworks based on the same mixed ligands, namely {[M(BTBD)2(bptc)0.5]·H2O}n (M = Zn (JXUST-48) and Cd (JXUST-49), BTBD = 4,7-bis(1H-1,2,4-triazol-1-yl)-2,1,3-benzothiadiazole, H4bptc = 3,3′,5,5′-biphenyltetracarboxylic acid), have been successfully constructed using solvothermal methods. Both remain stable when immersed in organic solvents and aqueous solutions with varying pH values of 1–14 for 24 h. Interestingly, JXUST-48 can detect 2,6-pyridine dicarboxylic acid (DPA) and thiamine (vitamin B1, VB1) by fluorescence enhancement and red-shift effect, whereas JXUST-49 is able to detect DPA and VB1 by fluorescence enhancement effect. The practical applicability of the sensors was validated using spiked recovery experiments, achieving recovery rates between 96.6% and 105.5%. More importantly, JXUST-48 and JXUST-49 have good biocompatibility and low cytotoxicity, while simultaneously achieving the capacity for fluorescence imaging and sensing of exogenous DPA and VB1 in vivo in living cells. This work provides some guidance for the development of MOF-based fluorophores based on different metal ions.
{"title":"pH- and Solvent-Stable ZnII/CdII Metal–Organic Frameworks with 2,1,3-Benzothiadiazole Derivative for Turn-On Fluorescence Sensing Dipicolinic Acid and Thiamine in Living Cells","authors":"Ding-Gui Cai,Shufen Pan,Teng-Fei Zheng,Chen Cao,Zhao-Bo Hu,Yan Peng,Yongquan Wu,He-Rui Wen,Sui-Jun Liu","doi":"10.1021/acs.analchem.5c04490","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c04490","url":null,"abstract":"The rapid detection of specific biological substances and biomarkers has the potential to contribute to the early identification, diagnosis, and prevention of diseases. Two examples of structurally similar luminescent metal–organic frameworks based on the same mixed ligands, namely {[M(BTBD)2(bptc)0.5]·H2O}n (M = Zn (JXUST-48) and Cd (JXUST-49), BTBD = 4,7-bis(1H-1,2,4-triazol-1-yl)-2,1,3-benzothiadiazole, H4bptc = 3,3′,5,5′-biphenyltetracarboxylic acid), have been successfully constructed using solvothermal methods. Both remain stable when immersed in organic solvents and aqueous solutions with varying pH values of 1–14 for 24 h. Interestingly, JXUST-48 can detect 2,6-pyridine dicarboxylic acid (DPA) and thiamine (vitamin B1, VB1) by fluorescence enhancement and red-shift effect, whereas JXUST-49 is able to detect DPA and VB1 by fluorescence enhancement effect. The practical applicability of the sensors was validated using spiked recovery experiments, achieving recovery rates between 96.6% and 105.5%. More importantly, JXUST-48 and JXUST-49 have good biocompatibility and low cytotoxicity, while simultaneously achieving the capacity for fluorescence imaging and sensing of exogenous DPA and VB1 in vivo in living cells. This work provides some guidance for the development of MOF-based fluorophores based on different metal ions.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"32 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139075","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}
Sensitive and specific detection of molecular biomarkers is fundamental to clinical diagnostics and biomedical research, yet existing CRISPR-based assays often suffer from nonspecific activation and cross-reactivity. Here, we introduce an asymmetric stem-loop translator that markedly enhances the specificity of a one-pot isothermal strand displacement amplification (SDA)-CRISPR/Cas12a assay. The asymmetric probe enables precise molecular recognition and controlled signal transduction, converting diverse targets into amplifiable DNA intermediates. Within the integrated one-tube system, SDA-generated DNA products directly trigger Cas12a trans-cleavage, yielding rapid fluorescence responses without thermal cycling or manual intervention. Systematic optimization of reaction parameters effectively mitigated enzyme inhibition and aerosol contamination. The resulting assay achieves highly specific and sensitive detection of miRNA with a detection limit of 500 fM, accurately distinguishing single- and double-base mutations. Owing to its modular design, the asymmetric stem-loop translator is readily adaptable to other analytes. By coupling to an aptamer-based recognition element, the system enables sensitive aflatoxin B1 detection. This work establishes a generalizable framework for enhancing CRISPR/Cas12a specificity through asymmetric molecular translation, offering a versatile platform for rapid nucleic acid and small-molecule diagnostics in clinical and point-of-care settings.
{"title":"An Asymmetric Stem-Loop Translator Enhances Specificity of One-Pot Isothermal CRISPR/Cas12a Assay","authors":"Liangyun Bu,Aijiao Yuan,Minqiao Zhao,Bo Pang,Juan Li,Yingxu Shang,Wenjing Xie,Hanyong Peng","doi":"10.1021/acs.analchem.5c07049","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07049","url":null,"abstract":"Sensitive and specific detection of molecular biomarkers is fundamental to clinical diagnostics and biomedical research, yet existing CRISPR-based assays often suffer from nonspecific activation and cross-reactivity. Here, we introduce an asymmetric stem-loop translator that markedly enhances the specificity of a one-pot isothermal strand displacement amplification (SDA)-CRISPR/Cas12a assay. The asymmetric probe enables precise molecular recognition and controlled signal transduction, converting diverse targets into amplifiable DNA intermediates. Within the integrated one-tube system, SDA-generated DNA products directly trigger Cas12a trans-cleavage, yielding rapid fluorescence responses without thermal cycling or manual intervention. Systematic optimization of reaction parameters effectively mitigated enzyme inhibition and aerosol contamination. The resulting assay achieves highly specific and sensitive detection of miRNA with a detection limit of 500 fM, accurately distinguishing single- and double-base mutations. Owing to its modular design, the asymmetric stem-loop translator is readily adaptable to other analytes. By coupling to an aptamer-based recognition element, the system enables sensitive aflatoxin B1 detection. This work establishes a generalizable framework for enhancing CRISPR/Cas12a specificity through asymmetric molecular translation, offering a versatile platform for rapid nucleic acid and small-molecule diagnostics in clinical and point-of-care settings.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"30 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139079","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}
Pub Date : 2026-02-09DOI: 10.1021/acs.analchem.5c07842
Baoping Zhu,Rui Han,Wenqing Wang,Shujie Cheng,Xiliang Luo
Electrochemical biosensors are susceptible to interference from biofouling of nonspecific proteins and the proteolytic hydrolysis of immobilized biomolecules by natural enzymes in complex biological media like blood. Therefore, enhancing the antifouling capability and the antienzymatic degradation ability is crucial for biosensors to perform in practical clinical samples. Herein, an electrochemical biosensing platform based on a multifunctional monocyclic peptide (MMCP) was developed, and the MMCP was designed with antifouling and antienzymatic degradation properties and the specific target (human epidermal growth factor receptor 2, HER2) recognition capability. The MMCP contains an antifouling part formed through covalent cyclization of a linear antifouling sequence (ECHHHHKHHHHCE) and a recognition d-peptide sequence (refffly) that can specifically recognize and bind HER2. Due to the cyclic structure and the adoption of d-amino acids, the MMCP can effectively resist the enzymatic degradation and shows significantly enhanced stability. The MMCP-based biosensor demonstrated outstanding antifouling performance and hydrolysis resistance properties in complex biological environments, and it was capable of sensitively detecting HER2 with a linear range of 1.0 pg mL–1 to 1.0 μg mL–1 and a low limit of detection of 0.35 pg mL–1. Additionally, the assay results of the biosensor for HER2 in clinical blood samples were consistent with those obtained from the enzyme-linked immunosorbent assay (ELISA). The strategy of designing a specific peptide reported herein offers an effective way to construct robust biosensors for biomarker detection in complicated clinical samples.
{"title":"A Robust Nonfouling Biosensor Based on an Engineered Multifunctional Monocyclic Peptide for Electrochemical Detection of Biomarkers in Human Blood","authors":"Baoping Zhu,Rui Han,Wenqing Wang,Shujie Cheng,Xiliang Luo","doi":"10.1021/acs.analchem.5c07842","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07842","url":null,"abstract":"Electrochemical biosensors are susceptible to interference from biofouling of nonspecific proteins and the proteolytic hydrolysis of immobilized biomolecules by natural enzymes in complex biological media like blood. Therefore, enhancing the antifouling capability and the antienzymatic degradation ability is crucial for biosensors to perform in practical clinical samples. Herein, an electrochemical biosensing platform based on a multifunctional monocyclic peptide (MMCP) was developed, and the MMCP was designed with antifouling and antienzymatic degradation properties and the specific target (human epidermal growth factor receptor 2, HER2) recognition capability. The MMCP contains an antifouling part formed through covalent cyclization of a linear antifouling sequence (ECHHHHKHHHHCE) and a recognition d-peptide sequence (refffly) that can specifically recognize and bind HER2. Due to the cyclic structure and the adoption of d-amino acids, the MMCP can effectively resist the enzymatic degradation and shows significantly enhanced stability. The MMCP-based biosensor demonstrated outstanding antifouling performance and hydrolysis resistance properties in complex biological environments, and it was capable of sensitively detecting HER2 with a linear range of 1.0 pg mL–1 to 1.0 μg mL–1 and a low limit of detection of 0.35 pg mL–1. Additionally, the assay results of the biosensor for HER2 in clinical blood samples were consistent with those obtained from the enzyme-linked immunosorbent assay (ELISA). The strategy of designing a specific peptide reported herein offers an effective way to construct robust biosensors for biomarker detection in complicated clinical samples.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"9 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146139082","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}
Pub Date : 2026-02-08DOI: 10.1021/acs.analchem.5c07839
Chunmeng Li,Xiangjian Zheng,Shangshang Xie,Deyong Lin,Zitian Liu
Accurate and specific detection of circular RNAs (circRNAs) is critical for vascular biology research and the clinical diagnosis of diabetes, particularly diabetic angiopathies. A major challenge in circRNA detection stems from the presence of abundant linear RNA isoforms that share identical sequences with circRNAs except for the back-splice junction. To overcome this limitation, we developed a novel detection strategy based on dual catalytically deactivated Cas13a/crRNA (dCas13a/crRNA) complexes that simultaneously recognize both ends of the circRNA back-splice junction. This system initiates a proximity ligation-triggered rolling circle amplification (RCA) reaction, producing long single-stranded DNA with tandemly repeated functional sequences. By combining dual dCas13a-guided recognition with proximity-mediated RCA, our method achieves exceptional specificity, enabling direct circRNA detection in complex RNA backgrounds, including linear isoforms, without requiring RNase R pretreatment. Coupled with triple catalytic hairpin assembly amplification, the assay detects circRNA with a detection limit of 0.083 fM within 150 min. The high specificity and sensitivity of this dCas13a/crRNA complex recognition-induced exponential amplification platform were validated in complex biological samples, demonstrating its broad potential as a versatile tool for sequence-specific RNA analysis and biomarker development in both basic research and clinical diagnostics of diabetic vascular complications.
{"title":"Deoxyribonucleic Acid Looped-Ag Nanocluster-Based Fluorescent Sensor for Accurate and Sensitive Circular Ribonucleic Acid Analysis via Dual Catalytically Deactivated Cas13a/crRNA Recognition-Mediated Proximity Ligation","authors":"Chunmeng Li,Xiangjian Zheng,Shangshang Xie,Deyong Lin,Zitian Liu","doi":"10.1021/acs.analchem.5c07839","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07839","url":null,"abstract":"Accurate and specific detection of circular RNAs (circRNAs) is critical for vascular biology research and the clinical diagnosis of diabetes, particularly diabetic angiopathies. A major challenge in circRNA detection stems from the presence of abundant linear RNA isoforms that share identical sequences with circRNAs except for the back-splice junction. To overcome this limitation, we developed a novel detection strategy based on dual catalytically deactivated Cas13a/crRNA (dCas13a/crRNA) complexes that simultaneously recognize both ends of the circRNA back-splice junction. This system initiates a proximity ligation-triggered rolling circle amplification (RCA) reaction, producing long single-stranded DNA with tandemly repeated functional sequences. By combining dual dCas13a-guided recognition with proximity-mediated RCA, our method achieves exceptional specificity, enabling direct circRNA detection in complex RNA backgrounds, including linear isoforms, without requiring RNase R pretreatment. Coupled with triple catalytic hairpin assembly amplification, the assay detects circRNA with a detection limit of 0.083 fM within 150 min. The high specificity and sensitivity of this dCas13a/crRNA complex recognition-induced exponential amplification platform were validated in complex biological samples, demonstrating its broad potential as a versatile tool for sequence-specific RNA analysis and biomarker development in both basic research and clinical diagnostics of diabetic vascular complications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"5 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146138737","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}
Pub Date : 2026-02-08DOI: 10.1021/acs.analchem.5c07182
Yanke Zhai,Qixuan Mu,Zhiping Tian,Yibing Ji,Ruijun Li
Despite its critical importance in disease diagnosis and drug safety, the development of methods for highly sensitive and enantioselective recognition of histidine enantiomers in complex biological media remains a formidable challenge. Here, the Chiral Metal–Organic Frameworks (CMOFs) called MOF-TH-Fe have been developed through a postsynthetic modification strategy. Using PCN-224 from zirconium-based Metal–Organic Frameworks (MOFs) as a scaffold, l-tartaric acid and l-histidine (l-His) were introduced as chiral linkers and anchored Fe3+ as the peroxidase-like (POD-like) activity center. MOF-TH-Fe enabled the chiral recognition of histidine enantiomers via dual-mode enantioselective detection. The results indicated that l-His significantly quenched the fluorescence intensity of MOF-TH-Fe at 655 nm, while d-His showed a negligible effect. Alternatively, a colorimetric method was established by exploiting the differential effect of l/d-His on the POD-like activity of MOF-TH-Fe. The fluorescence and colorimetric assays offered detection limits of 0.57 and 1.74 μM, respectively. Molecular simulations explained the mechanism of chiral recognition and revealed the different interactions between chiral linkers and histidine enantiomers. To verify its potential in chiral separation, MOF-TH-Fe was integrated with a fixed-bed continuous flow system to separate the enantiomers of histidine. After 3.5 h of continuous operation, an enantiomeric excess (ee) of 71.69% was achieved. This study successfully established a multifunctional platform integrating chiral recognition, separation, and catalysis, providing design insights for the development of next-generation multifunctional chiral nanomaterials.
{"title":"Integrated Chiral Recognition and Enantiomeric Separation of Histidine by Metal–Organic Frameworks Engineered with Chiral Linkers","authors":"Yanke Zhai,Qixuan Mu,Zhiping Tian,Yibing Ji,Ruijun Li","doi":"10.1021/acs.analchem.5c07182","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c07182","url":null,"abstract":"Despite its critical importance in disease diagnosis and drug safety, the development of methods for highly sensitive and enantioselective recognition of histidine enantiomers in complex biological media remains a formidable challenge. Here, the Chiral Metal–Organic Frameworks (CMOFs) called MOF-TH-Fe have been developed through a postsynthetic modification strategy. Using PCN-224 from zirconium-based Metal–Organic Frameworks (MOFs) as a scaffold, l-tartaric acid and l-histidine (l-His) were introduced as chiral linkers and anchored Fe3+ as the peroxidase-like (POD-like) activity center. MOF-TH-Fe enabled the chiral recognition of histidine enantiomers via dual-mode enantioselective detection. The results indicated that l-His significantly quenched the fluorescence intensity of MOF-TH-Fe at 655 nm, while d-His showed a negligible effect. Alternatively, a colorimetric method was established by exploiting the differential effect of l/d-His on the POD-like activity of MOF-TH-Fe. The fluorescence and colorimetric assays offered detection limits of 0.57 and 1.74 μM, respectively. Molecular simulations explained the mechanism of chiral recognition and revealed the different interactions between chiral linkers and histidine enantiomers. To verify its potential in chiral separation, MOF-TH-Fe was integrated with a fixed-bed continuous flow system to separate the enantiomers of histidine. After 3.5 h of continuous operation, an enantiomeric excess (ee) of 71.69% was achieved. This study successfully established a multifunctional platform integrating chiral recognition, separation, and catalysis, providing design insights for the development of next-generation multifunctional chiral nanomaterials.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"42 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2026-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146138736","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}
Pub Date : 2026-02-07DOI: 10.1021/acs.analchem.5c06031
Zhihao Zhou, Kangrui Hu, Nan Jia, Zhongbo Wang, Ke Ding, Qi Chen, Yiyue Yan, Lin Su, Xiayu Qian, Xin Fan, Lin Xie, Guangji Wang, Tengjie Yu, Yan Liang
Mass spectrometry imaging (MSI) can directly detect surface analytes, enabling spatial mapping of metabolite synthesis and migration within tissues. However, MSI often lacks the sensitivity for routine analysis of low exposure, low molecular weight endogenous metabolites. Amine-containing metabolites, such as methylamine, dimethylamine, trimethylamine, lysine, dopamine, and 5-hydroxytryptamine, are widely used as medical biomarkers due to their potential biological activity or toxicity. Herein, we innovatively developed a universal in situ derivatization workflow combined with desorption electrospray ionization MS/MS (DESI-MS/MS) for mapping endogenous amine-containing metabolites, with trimethylamine (TMA) selected as a case study. For the first time, tert-butyl bromoacetate (TBBA) was used as an in situ derivatization reagent, which improved sensitivity and reproducibility by overcoming the strong matrix effect and signal instability of low molecular weight compounds in MS analysis. Our data suggested that the derivatization efficiency was affected by the alkalinity of the tissue surface, while spray/extraction solvent significantly affected the sensitivity for TMA-TBBA. Notably, DESI-MS/MS exhibited good linearity, reproducibility, and stability in the analysis of amine-containing metabolites derived from TBBA. The optimized workflow was utilized for the visual analysis of endogenous TMA distribution in the brain and serum of mice, revealing the enrichment characteristics of TMA in the cortex and hippocampus, as well as the severe accumulation of TMA in aged animals. Therefore, this targeted DESI-MS/MS-MSI approach, combined with in situ derivatization, offers novel avenues for the routine quantitation of intratissue amine-containing metabolites. In principle, this assay can be extended to a wide variety of metabolites in different biological samples.
{"title":"In Situ Derivatization Combined with DESI-MRM-MS/MS for Spatial Mapping Intratissue Amine-Containing Metabolites, Trimethylamine as a Case.","authors":"Zhihao Zhou, Kangrui Hu, Nan Jia, Zhongbo Wang, Ke Ding, Qi Chen, Yiyue Yan, Lin Su, Xiayu Qian, Xin Fan, Lin Xie, Guangji Wang, Tengjie Yu, Yan Liang","doi":"10.1021/acs.analchem.5c06031","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c06031","url":null,"abstract":"<p><p>Mass spectrometry imaging (MSI) can directly detect surface analytes, enabling spatial mapping of metabolite synthesis and migration within tissues. However, MSI often lacks the sensitivity for routine analysis of low exposure, low molecular weight endogenous metabolites. Amine-containing metabolites, such as methylamine, dimethylamine, trimethylamine, lysine, dopamine, and 5-hydroxytryptamine, are widely used as medical biomarkers due to their potential biological activity or toxicity. Herein, we innovatively developed a universal in situ derivatization workflow combined with desorption electrospray ionization MS/MS (DESI-MS/MS) for mapping endogenous amine-containing metabolites, with trimethylamine (TMA) selected as a case study. For the first time, <i>tert</i>-butyl bromoacetate (TBBA) was used as an in situ derivatization reagent, which improved sensitivity and reproducibility by overcoming the strong matrix effect and signal instability of low molecular weight compounds in MS analysis. Our data suggested that the derivatization efficiency was affected by the alkalinity of the tissue surface, while spray/extraction solvent significantly affected the sensitivity for TMA-TBBA. Notably, DESI-MS/MS exhibited good linearity, reproducibility, and stability in the analysis of amine-containing metabolites derived from TBBA. The optimized workflow was utilized for the visual analysis of endogenous TMA distribution in the brain and serum of mice, revealing the enrichment characteristics of TMA in the cortex and hippocampus, as well as the severe accumulation of TMA in aged animals. Therefore, this targeted DESI-MS/MS-MSI approach, combined with in situ derivatization, offers novel avenues for the routine quantitation of intratissue amine-containing metabolites. In principle, this assay can be extended to a wide variety of metabolites in different biological samples.</p>","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":" ","pages":""},"PeriodicalIF":6.7,"publicationDate":"2026-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146130582","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}
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