Rapid Communications in Mass Spectrometry最新文献_第2页

UHPLC-Q-Orbitrap HRMS-Based Machine Learning Constructs the Integrated Biomarker Profiling of Type 2 Diabetes and Diabetic Heart Disease 基于UHPLC-Q-Orbitrap hrms的机器学习构建2型糖尿病和糖尿病性心脏病的综合生物标志物谱

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-30 DOI: 10.1002/rcm.70044

Jianglan Long, Yueyue Wang, Qiushi Fang, Zhe Shi, Zhenzhen Wang, Dan Yan

Rationale

Over 70% of diabetic patients die from cardiovascular disease, in which diabetic heart disease (DHD) is an important cause of death in individuals with type 2 diabetes (T2D). It is hence imperative to explore the simple, rapid, and economical method for diagnosing DHD from T2D.

Methods

T2D and DHD patients were recruited, and their serum samples were used for metabolomic analysis to identify differential metabolites. Logistic regression analysis and receiver operating characteristic curve analysis were performed to identify candidate biomarkers. Moreover, four machine learning methods were used to construct the integrated biomarker profiling (IBP) models with the candidate biomarkers. Gini impurity was employed to select characteristic candidate biomarkers.

Results

Eighty-four differential metabolites were identified in the serum of 58 T2D and 62 DHD patients. Logistic regression analysis indicated that 17 differential metabolites were protective factors, whereas 39 were risk factors for DHD. Further, 29 differential metabolites were identified as the candidate biomarkers of DHD after receiver operating characteristic curve analysis. After comparing the predictive performance of the four machine learning models, the IBP was constructed based on the eXtreme Gradient Boosting (XGBoost) with six candidate biomarkers, which were sphingomyelin (d18:0/16:1), deoxycholic acid, hexadecanedioic acid, phosphatidylcholine (20:5/18:3), L-tryptophan, and N-undecanoylglycine from the ranked results of Gini impurity. The accuracy of the IBP for distinguishing T2D and DHD reached 88.89%, with a 100% accuracy in predicting DHD from T2D patients.

Conclusions

The IBP, composed of six metabolites, can effectively predict DHD from T2D, and it is expected to become a screening indicator for early-stage DHD.

理由：超过70%的糖尿病患者死于心血管疾病，其中糖尿病性心脏病（DHD）是2型糖尿病（T2D）患者死亡的重要原因。因此，探索从T2D诊断DHD的简便、快速、经济的方法势在必行。方法：招募T2D和DHD患者，进行血清代谢组学分析，鉴别差异代谢物。采用Logistic回归分析和受试者工作特征曲线分析确定候选生物标志物。此外，利用四种机器学习方法构建了候选生物标志物的综合生物标志物分析（IBP）模型。采用基尼杂质筛选特征候选生物标志物。结果：在58例T2D和62例DHD患者血清中鉴定出84种差异代谢物。Logistic回归分析显示17种差异代谢物是DHD的保护因素，39种差异代谢物是DHD的危险因素。此外，通过受试者工作特征曲线分析，确定了29种差异代谢物作为DHD的候选生物标志物。在比较了四种机器学习模型的预测性能后，基于极限梯度增强（XGBoost）构建了IBP，其中六个候选生物标志物是鞘磷脂（d18:0/16:1）、脱氧胆酸、十六烷二酸、磷脂酰胆碱（20:5/18:3）、l -色氨酸和n -十一烷酰甘氨酸，这些生物标志物来自Gini杂质排名结果。IBP区分T2D和DHD的准确率达到88.89%，预测T2D患者的DHD准确率为100%。结论：由6种代谢物组成的IBP可有效预测T2D的DHD，有望成为早期DHD的筛查指标。

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引用次数: 0

Comprehensive Characterization of the Chemical Constituents, Serum Pharmacochemistry, and Quality Control of Jieyu Pills Utilizing UHPLC-Orbitrap Fusion MS UHPLC-Orbitrap融合质谱法综合表征解郁丸的化学成分、血清药物化学及质量控制。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-30 DOI: 10.1002/rcm.70042

Lingyu Ye, Shuding Sun, Ting Wang, Jia Song, Li Wang, Yufang Miao, Di Zhao, Qi Sun, Yan Wan, Suxiang Feng

Rationale

Depression is currently the third leading cause of disease burden worldwide by 2030. Jieyu Pills (JYP) comprise 10 herbs demonstrated clinical effectiveness in managing depression with low by-effects. Owing to the complexity of herbal compound formulations, identifying chemical constituents is insufficient to discover the specificity and correlation between quality and therapeutic efficacy. Thus, selecting and detecting appropriate quality control (QC) markers for herbal medicines remains a challenge.

Methods

This study has been screened and evaluated appropriate QC markers through a correlation analysis of “ingredient–pharmacological efficacy.” Firstly, chemical components of JYP were systematically characterized in vivo and in vitro using UHPLC-Orbitrap Fusion MS. Then, network pharmacology analysis was conducted to predict potential active components of JYP. Finally, core effectors of JYP were screened out as QC markers and subjected to targeted quantitative analysis.

Results

A total of 188 compounds were identified in vitro, and 47 prototype constituents plus 41 metabolites were detected in serum. Network analysis revealed 13 core targets such as AKT1 and TNF enriched in inflammation, apoptosis, and AGE-RAGE signaling pathways, aligning with known depression mechanisms. Integrating serum-exposed components with network results highlighted paeoniflorin, gallic acid, liquiritin, and their metabolites as central modulators of neuroinflammatory and neuroprotective processes. Based on this “chemical entity–metabolic fate–target–pathway” framework, 20 compounds were selected as potential QC markers, and quantitative analysis confirmed high batch consistency.

Conclusions

This study provides the first integrated in vitro–in vivo chemical and mechanistic profiling of JYP, elucidates its metabolic characteristics and core antidepressant pathways, and establishes scientifically grounded QC markers. The research provides a novel perspective for investigating complex herbal prescriptions.

理由：到2030年，抑郁症目前是全球疾病负担的第三大原因。解郁丸（JYP）由10种草药组成，在治疗抑郁症方面具有临床效果，副作用低。由于中药复方的复杂性，确定化学成分不足以发现质量与疗效之间的特异性和相关性。因此，选择和检测合适的中草药质量控制（QC）标记仍然是一个挑战。方法：通过“成分-药效”的相关性分析，筛选和评价合适的QC标志物。首先采用UHPLC-Orbitrap融合质谱法对JYP的体内外化学成分进行系统表征，然后进行网络药理学分析，预测JYP的潜在有效成分。最后，筛选JYP的核心效应物作为QC标记，进行有针对性的定量分析。结果：体外共鉴定出188种化合物，血清中检测到47种原型成分和41种代谢物。网络分析揭示了13个核心靶点，如AKT1和TNF在炎症、凋亡和AGE-RAGE信号通路中富集，与已知的抑郁机制一致。将血清暴露成分与网络结果相结合，强调芍药苷、没食子酸、甘草素及其代谢物是神经炎症和神经保护过程的中枢调节剂。基于这一“化学实体-代谢命运-目标途径”的框架，选择了20个化合物作为潜在的QC标记物，定量分析证实了高批次一致性。结论：本研究首次提供了JYP的体内外化学和机制分析，阐明了其代谢特征和核心抗抑郁途径，建立了科学依据的QC标记。该研究为复杂中药处方的研究提供了新的视角。

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引用次数: 0

Minimally Destructive Radiocarbon Dating of Bone 骨的最小破坏性放射性碳定年法。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-30 DOI: 10.1002/rcm.70040

Tom Higham, Katharina Luftensteiner, Laura van der Sluis, Maddalena Giannì, Maria Wiegele, Anastasia Papadogianni, Peter Steier, Daniela Gruber, Brigitte Schmidt, Katy Schmidt, Andrei Belinski, Romain Mensan, Maxim Kozlikin, Michael Shunkov, Petr Neruda, Zdeňka Nerudová, Barbara Horejs, John Schulze, Katerina Douka, Thomas W. Stafford Jr.

Rationale

Bone is commonly used in radiocarbon dating in archaeology and other disciplines. Despite advances in collagen extraction protocols, the process remains destructive, requiring sawing, drilling or crushing of bone material. While non-destructive approaches have recently been applied in ancient genomics and palaeoproteomics, no equivalent approach has been established for radiocarbon dating of bone. We explored whether this is possible using a series of experiments.

Methods

We experimented by using a water-based approach to extract soluble collagen from whole bone and teeth samples. We heated the samples in hot (75°C and 90°C) water for several hours. We obtained the soluble collagen fraction of the bone and purified and AMS dated the extracts. We used standard reference bones and samples from archaeological sites.

Results

We found that the amino acid composition, C/N atomic ratios, δ¹³C and δ¹⁵N values of the hot-water-extracted soluble collagen were comparable to collagen isolated from the same bones using classic Longin collagen methods. Bone and teeth from Bronze Age and Middle and Upper Paleolithic sites, which had been dated previously using routine destructive methods that involved acid demineralization, yielded dates on the water-soluble fraction that were in good agreement with these earlier results.

Conclusions

We show that a minimally destructive collagen extraction, coupled with an additional purification step such as ultrafiltration or XAD-2 purification, yields identical radiocarbon ages to those obtained via the routine destructive methods, but without any visible external damage. The method may allow us in future to date precious artefacts, ornaments and museum objects without significant alteration.

理论依据：骨头通常用于考古学和其他学科的放射性碳测年。尽管胶原蛋白提取技术有所进步，但这一过程仍然具有破坏性，需要锯切、钻孔或粉碎骨骼材料。虽然非破坏性方法最近已应用于古代基因组学和古蛋白质组学，但尚未建立用于骨骼放射性碳定年的等效方法。我们通过一系列实验来探索这是否可能。方法：采用水基法提取全骨和全牙标本中的可溶性胶原蛋白。我们将样品在热水（75°C和90°C）中加热几个小时。我们获得了骨的可溶性胶原蛋白部分，并对其进行了纯化和AMS定年。我们使用了标准的参考骨骼和考古遗址的样本。结果：热水提取的可溶性胶原蛋白的氨基酸组成、C/N原子比、δ13C和δ15N值与经典Longin胶原法分离的胶原蛋白相当。来自青铜器时代和旧石器时代中晚期遗址的骨头和牙齿，以前是用常规的破坏性方法测定年代的，其中包括酸脱矿，得出的水溶性部分的日期与这些早期的结果非常吻合。结论：我们证明了一种破坏性最小的胶原蛋白提取，加上额外的纯化步骤，如超滤或XAD-2纯化，产生的放射性碳年龄与通过常规破坏性方法获得的放射性碳年龄相同，但没有任何可见的外部损伤。这种方法可以让我们在未来确定珍贵的人工制品、装饰品和博物馆物品的年代，而不会有明显的改变。

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引用次数: 0

Phospholipid Profiling Established by Structure-Rich Fragments for Molecular Species Level Shotgun Analysis 用富结构片段建立磷脂谱图，用于分子种水平鸟枪分析。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-27 DOI: 10.1002/rcm.70038

Rong Chen, Amber H. Jannasch, Bruce R. Cooper, Jonathan H. Shannahan, Christina R. Ferreira

Rationale

Accurate identification of phospholipid molecular species remains a major challenge in shotgun lipidomics because conventional tandem mass spectrometry typically resolves only one structural moiety at a time. This structural ambiguity limits confident lipid biomarker discovery and biological interpretation. Improving structural specificity without sacrificing analytical speed is therefore critical for lipidomics and disease-related studies.

Methods

Electrospray ionization tandem mass spectrometry was performed using direct infusion on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode. MRMs were designed based on structure-rich phospholipid fragments containing both the headgroup and one fatty acyl chain. Lipids were extracted from mouse liver and brain tissues and analysed without chromatographic separation, and normal-phase LC was used for lipid headgroup confirmation only.

Results

Structure-rich MS/MS transitions enabled molecular species identification of both diacyl and ether phospholipids. 15 PUFA-containing phospholipids were identified as candidate biomarkers differentiating healthy and metabolic syndrome mouse livers, revealing opposing regulation among structurally similar species supported by complementary fragmentation and LC evidence. In mouse brains, three ether lipid biomarkers were discovered, including plasmalogens and plasmanyl lipids, with distinct disease-associated trends.

Conclusion

This study demonstrates that structure-rich MS/MS transitions substantially improve phospholipid structural specificity in shotgun lipidomics while maintaining high throughput. The method enables reliable identification of individual lipid species with minimal isomer interference and is readily compatible with existing workflows. This strategy offers a practical path toward more precise lipid biomarker discovery and mechanistic insight into metabolic disease.

原理：由于传统的串联质谱法通常一次只能解析一个结构片段，因此准确鉴定磷脂分子种类仍然是霰弹枪脂质组学的主要挑战。这种结构上的模糊性限制了脂质生物标志物的发现和生物学解释。因此，在不牺牲分析速度的情况下提高结构特异性对于脂质组学和疾病相关研究至关重要。方法：采用多反应监测（MRM）模式运行的三重四极杆质谱仪，直接输注电喷雾电离串联质谱法。MRMs是基于含有头基和一个脂肪酰基链的结构丰富的磷脂片段设计的。从小鼠肝脏和脑组织中提取脂质，不进行色谱分离分析，仅使用正相LC进行脂质头组确认。结果：结构丰富的MS/MS转换使二酰基磷脂和醚磷脂的分子种类鉴定成为可能。15种含pufa的磷脂被确定为区分健康和代谢综合征小鼠肝脏的候选生物标志物，揭示了结构相似的物种之间的相反调控，这得到了互补碎片化和LC证据的支持。在小鼠大脑中，发现了三种乙醚类脂质生物标志物，包括浆磷脂原和浆磷脂酰脂质，它们具有明显的疾病相关趋势。结论：本研究表明，富结构的MS/MS转换在保持高通量的同时，显著提高了霰弹枪脂质组学的磷脂结构特异性。该方法能够以最小的同分异构体干扰可靠地鉴定单个脂类，并且易于与现有工作流程兼容。这一策略为更精确地发现脂质生物标志物和了解代谢疾病的机制提供了一条实用的途径。

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引用次数: 0

In Vitro Metabolism and Analytical Characterization of SLU-PP-332 and SLU-PP-915: Novel Pan-ERR Agonists With Doping Potential SLU-PP-332和SLU-PP-915：具有兴奋剂潜力的新型泛err激动剂的体外代谢和分析表征

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-26 DOI: 10.1002/rcm.70039

Tristan Möller, Oliver Krug, Mario Thevis

Rationale

Estrogen-related receptor (ERR) agonists such as the drug candidates SLU-PP-332 and SLU-PP-915 are currently being investigated as exercise mimetics, given their ability to trigger human physiological processes similar to those initiated by actual physical activity. This capability prompted the consideration of these compounds as drugs potentially relevant for sports drug testing programs.

Methods

The two pan-ERR agonists SLU-PP-332 and SLU-PP-915 were characterized using liquid chromatography-high resolution (tandem) mass spectrometry (LC–HRMS/MS). Furthermore, the in vitro metabolic transformation products of both compounds prepared by means of human liver S9 fraction (S9 fraction) and human liver microsomes (HLMs) were analyzed. In addition, selected metabolites of SLU-PP-915 were synthesized and their structures were analyzed by nuclear magnetic resonance (NMR) spectroscopy.

Results

A total of nine metabolites were identified for SLU-PP-332, consisting of six Phase-I metabolites and three Phase-II conjugates. Conversely, the analysis of SLU-PP-915 yielded only Phase-I transformation products, with a total of seven metabolites identified. In both cases, an in-depth structural elucidation was conducted to obtain a comprehensive overview of the detected metabolites. Furthermore, three metabolites of SLU-PP-915 were confirmed through chemical synthesis and NMR.

Conclusion

The results obtained in this study gave an in-depth view into the analysis and in vitro metabolism of the newly developed pan-ERR agonists SLU-PP-332 and SLU-PP-915. This may help to uncover the illicit use of these novel compounds as potential performance-enhancing substances.

理由：雌激素相关受体（ERR）激动剂，如候选药物SLU-PP-332和SLU-PP-915，目前正在作为运动模拟剂进行研究，因为它们具有触发人体生理过程的能力，类似于实际体育活动所启动的生理过程。这种能力促使人们考虑将这些化合物作为运动药物测试项目的潜在相关药物。方法：采用液相色谱-串联质谱（LC-HRMS/MS）对两种泛err激动剂SLU-PP-332和SLU-PP-915进行表征。进一步分析了人肝脏S9组分（S9 fraction）和人肝脏微粒体（HLMs）制备的两种化合物的体外代谢转化产物。此外，合成了SLU-PP-915的代谢物，并用核磁共振（NMR）对其结构进行了分析。结果：SLU-PP-332共鉴定出9个代谢物，包括6个i期代谢物和3个ii期偶联物。相反，对SLU-PP-915的分析只得到i期转化产物，共鉴定出7种代谢物。在这两种情况下，进行了深入的结构解析，以获得检测到的代谢物的全面概述。此外，通过化学合成和核磁共振证实了SLU-PP-915的三个代谢物。结论：本研究结果对新开发的泛err激动剂SLU-PP-332和SLU-PP-915的分析和体外代谢有了深入的了解。这可能有助于揭露这些新型化合物作为潜在的性能增强物质的非法使用。

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引用次数: 0

Colicin-Bearing Plasmids Carried by Shiga Toxin-Producing E. coli (STEC) Analyzed by Targeted Top-Down MS Analysis 产志贺毒素大肠杆菌（STEC）携带大肠杆菌素质粒的定向自上而下质谱分析

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-23 DOI: 10.1002/rcm.70031

Clifton K. Fagerquist, Yanlin Shi, Mahesh Koirala, Maria T. Brandl

Rationale

Colicins are protein toxins produced by bacteria that destroy the nucleic acids or outer membranes of their bacterial neighbors. Their genes are encoded in small extra-chromosomal plasmids that play an important role in bacterial survival. Although colicins do not contribute to bacterial pathogen virulence, it is important to develop methods to identify molecular determinants that facilitate pathogen survival.

Methods

Shiga toxin-producing Escherichia coli (STEC) were analyzed for the presence of colicin-bearing plasmids using antibiotic induction, MALDI-TOF-TOF, Orbitrap mass spectrometry, and targeted top-down analysis using in-house software. Small plasmid sequencing was used to confirm plasmid-encoded genes as well as their upstream regulation. AlphaFold3 was used to rationalize expected (as well as anomalous) fragment ions detected by MS/MS post-source decay (PSD).

Results

Colicin immunity (Imm) proteins were detected and identified by targeted top-down mass spectrometry in two STEC strains (serotypes O26:H11 and O104:H7) that carried one or more colicin plasmids. ImBac, ImmD, and a 7.8 kDa hypothetical protein (whose gene resides on a plasmid that encodes a pore-forming colicin) were detected and identified. Whole genome sequencing (by other groups) and our small plasmid sequencing confirmed the colicin/immunity genes as well as their upstream regulatory control.

Conclusions

MALDI-TOF-TOF-MS/MS-PSD is an effective platform for rapid detection and identification of inducible antibacterial protein toxins. We also note that the previously reported glycine-enhanced aspartic acid effect (AAE) appears to occur most often at unstructured/linker regions of the polypeptide backbone.

原理：粘菌素是细菌产生的蛋白质毒素，它能破坏邻近细菌的核酸或外膜。它们的基因编码在小的染色体外质粒中，在细菌存活中起重要作用。虽然粘菌素不促进细菌病原体的毒力，但重要的是要发展方法来确定促进病原体生存的分子决定因素。方法：采用抗生素诱导法、MALDI-TOF-TOF、Orbitrap质谱法和内部软件进行定向自顶向下分析，分析产志贺毒素大肠杆菌（STEC）携带大肠杆菌素质粒的存在。小质粒测序用于确认质粒编码的基因及其上游调控。使用AlphaFold3对MS/MS后源衰减（PSD）检测到的预期（以及异常）片段离子进行合理化。结果：用自上而下定向质谱法检测并鉴定了携带一种或多种大肠杆菌素质粒的两种产志在大肠杆菌毒素菌株（O26:H11和O104:H7血清型）的大肠杆菌素免疫（Imm）蛋白。检测并鉴定了ImBac、imd和一个7.8 kDa的假设蛋白（其基因位于编码成孔粘菌素的质粒上）。全基因组测序（由其他团队）和我们的小质粒测序证实了colicin/免疫基因及其上游调控。结论：MALDI-TOF-TOF-MS/MS-PSD是一种快速检测和鉴定诱导型抗菌蛋白毒素的有效平台。我们还注意到，先前报道的甘氨酸增强天冬氨酸效应（AAE）似乎最常发生在多肽主链的非结构化/连接区域。

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引用次数: 0

Label-Free Quantitative Proteomics Analysis Revealed the Peptide and Keratin Protein Pattern in Different Types of Sunda Porcupine (Hystrix javanica) Quill 无标记定量蛋白质组学分析揭示了不同类型巽他豪猪（Hystrix javanica）羽毛的肽和角蛋白模式。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-22 DOI: 10.1002/rcm.70036

Andhika Yudha Prawira, Maharani Kartika Ramadhan, Tulus Maulana, Ni Luh Putu Rischa Phadmacanty, Rizki Fitrawan Yuneldi, Srihadi Agungpriyono, Berry Juliandi

Rationale

Sunda porcupine (Hystrix javanica) has quills that exhibit considerable morphological diversity and functionality, yet the molecular variation underlying these differences remains insufficiently explored. Comprehensive proteomic profiling provides an opportunity to examine peptide composition and keratin expression patterns across quill types. This study aims to characterize molecular distinctions among Sunda porcupine quills using label-free quantitative proteomic analysis.

Methods

Three quill types—spine, true quill, and rattle quill—were collected, cleaned, pulverized, and subjected to keratin-specific extraction employing reducing agents. Extracted proteins underwent in-solution digestion before analysis using LC–MS/MS on a high-resolution Orbitrap system. Peptide and protein identification utilized SequestHT against curated rodent keratin databases. Data were processed through multivariate statistical analyses, including PCA, PLS-DA, SOM, and heatmap clustering to assess quill-specific clustering, peptide distribution, and keratin profile variation among quill types.

Results

LC–MS/MS identified 653 peptides and 70 homologous proteins, revealing molecular variation among quill types. True quill and spine displayed overlapping peptide abundance patterns, whereas rattle quill demonstrated distinct clustering. Amino acid composition varied among quills, reflecting structural and functional differentiation. True quill and spine showed higher intensity of proteins than rattle quill. Keratin type I cuticular and cytoskeletal proteins were the most matched proteins. Protein profile indicated high similarity to keratins from Hystricomorpha rodent species.

Conclusion

The study demonstrates clear molecular differentiation among quill types, with true quill and spine exhibiting closer proteomic similarity than rattle quill. Distinct peptides identified in each quill category highlight their potential as biomarkers for quill-type discrimination, although further validation is required to confirm their diagnostic reliability.

理由：巽他豪猪（Hystrix javanica）的刺具有相当大的形态多样性和功能，但这些差异背后的分子变异仍未得到充分的探索。全面的蛋白质组学分析提供了一个机会来检查多肽组成和角蛋白表达模式的羽毛类型。本研究旨在利用无标记定量蛋白质组学分析表征巽他豪猪刺的分子差异。方法：收集棘毛、真毛和摇毛三种羽毛，清洗、粉碎，用还原剂进行角蛋白特异性提取。在高分辨率Orbitrap系统上使用LC-MS/MS进行分析之前，提取的蛋白质在溶液中消化。利用SequestHT对整理的啮齿动物角蛋白数据库进行肽和蛋白鉴定。数据通过多元统计分析进行处理，包括PCA、PLS-DA、SOM和热图聚类，以评估羽毛的特异性聚类、肽分布和角蛋白谱在羽毛类型之间的变化。结果：LC-MS/MS鉴定出653个多肽和70个同源蛋白，揭示了不同羽毛笔类型间的分子变异。真羽和棘羽显示重叠的肽丰度模式，而摇羽显示明显的聚类。不同刺间氨基酸组成不同，反映了结构和功能的差异。真羽和棘羽的蛋白质强度高于摇羽。角蛋白I型与角质层蛋白和细胞骨架蛋白匹配度最高。蛋白谱显示与水形目啮齿类动物的角蛋白高度相似。结论：研究结果表明，真羽和棘羽在蛋白质组学上具有明显的相似性。尽管需要进一步验证以确认其诊断可靠性，但在每个羽毛笔类别中鉴定出的不同肽突出了它们作为羽毛笔类型区分的生物标志物的潜力。

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引用次数: 0

A Novel 1,3-Sigmatropic Rearrangement in an Iminium Ion Generated in Collision-Induced Decomposition During Electrospray Ionization High Resolution Multistage Mass Spectrometry 在电喷雾电离高分辨率多级质谱分析中碰撞诱发分解产生的一种新的1,3-异位重排。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-21 DOI: 10.1002/rcm.70037

Wenbin Chen, Jiangbo Xi, Zhiyuan Gao, Juan Zhang, Junjun Zhang, Zhengwu Bai, Min Li

Rationale

Rapid structural elucidation of unknown pharmaceutical impurities is critical throughout the whole life cycle of drug development. It can be achieved by using multistage mass spectrometric analysis, in conjunction with the understanding of dissociation mechanisms of the ions generated by electrospray ionization.

Methods

Liquid chromatography-UV/photo diode array detection—high resolution multistage mass spectrometric analysis (HiRes MSⁿ) was used to elucidate the structure of an impurity of silodosin. The structure was confirmed by 1D and 2D NMR spectroscopy.

Results

The impurity is identified as N-cyanomethylsilodosin, a solution degradant of silodosin. A novel 1,3-sigmatropic rearrangement was observed during one of its MS³ dissociation pathways. A mechanism is proposed for the 1,3-sigmatropic rearrangement, which is supported by the MSⁿ behavior of a ¹³C-isotope labeled derivative of N-cyanomethylsilodosin.

Conclusion

Elucidation of such an intra-molecular rearrangement mechanism not only enriches our understanding of the gas phase dissociation chemistry, but also should facilitate future rapid structural characterization of unknown pharmaceutical impurities and metabolites using multistage mass spectrometric analysis.

原理：在药物开发的整个生命周期中，未知药物杂质的快速结构解析是至关重要的。它可以通过使用多级质谱分析，结合对电喷雾电离产生的离子的解离机制的理解来实现。方法：采用液相色谱-紫外/光电二极管阵列检测-高分辨率多级质谱分析（HiRes MSn）对西洛多辛杂质进行结构分析。通过一维和二维核磁共振波谱分析证实了其结构。结果：该杂质经鉴定为n -氰甲基西洛多辛，是西洛多辛的溶液降解剂。在其MS3解离途径之一中观察到一种新的1,3-异位重排。通过13c同位素标记的n -氰甲基西洛多辛衍生物的MSn行为，提出了1,3-异位重排的机制。结论：这种分子内重排机制的阐明不仅丰富了我们对气相解离化学的理解，而且有助于未来利用多级质谱分析快速表征未知药物杂质和代谢物的结构。

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引用次数: 0

Mass Spectrometry-Based Proteomics to Understand Immune Thrombocytopenic Purpura: A Review 基于质谱的蛋白质组学研究免疫性血小板减少性紫癜：综述。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-20 DOI: 10.1002/rcm.70022

Syed Kashif Raza, Ayesha Iqbal

Background

Immune thrombocytopenic purpura (ITP) is an autoimmune bleeding disorder with substantial variability in bleeding manifestations and a generally low risk of fatal outcomes. Diagnosis is based on exclusion and is supported by thrombocytopenia in peripheral blood and characteristic megakaryocytic findings in bone marrow examination. In many adults, treatment remains suboptimal, and morbidity may result from both bleeding and the adverse effects of immunosuppressive therapies.

Rationale

Identifying the underlying disease process is essential for improving diagnostic precision, developing targeted therapies, and enhancing patient outcomes.

Objective

This review summarizes mass spectrometry-based proteomics studies investigating the pathogenesis of ITP and their diagnostic, prognostic, and therapeutic implications.

Conclusion

Despite significant clinical heterogeneity, proteomic research in ITP remains limited, underscoring the need for comprehensive molecular studies to better elucidate disease mechanisms.

背景：免疫性血小板减少性紫癜（ITP）是一种自身免疫性出血性疾病，出血表现有很大的变异性，通常死亡风险较低。诊断是基于排除和支持外周血血小板减少和特征性巨核细胞的骨髓检查结果。在许多成年人中，治疗仍然不够理想，并且发病率可能由出血和免疫抑制治疗的不良反应引起。基本原理：确定潜在的疾病过程对于提高诊断精度、开发靶向治疗和提高患者预后至关重要。目的：本文综述了基于质谱的蛋白质组学研究ITP的发病机制及其诊断、预后和治疗意义。结论：尽管存在显著的临床异质性，但ITP的蛋白质组学研究仍然有限，强调需要进行全面的分子研究以更好地阐明疾病机制。

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引用次数: 0

Pyrene-Conjugated, 2-Pyridinecarboxaldehyde Derivatives as N-Terminus-Specific Tags for MALDI- and LALDI-MS 吡啶共轭2-吡啶甲酸衍生物作为MALDI-和LALDI-MS的n端特异性标签

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2026-01-20 DOI: 10.1002/rcm.70034

Mujia Jenny Li, Murat Kucukdisli, Florian Braun, David W. Will, Nadine Meier, Bettina Wehrle, Larissa Chiara Meyer, Melanie Christine Föll, Oliver Schilling

Background

Proteolytic processing is a fundamental post-translational modification. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) workflows are powerful for degradomic analyses but inherently sacrifice spatial information, a critical aspect for investigating biological systems such as aberrant extracellular matrix remodeling and alterations of the tumor microenvironment. Matrix-assisted laser desorption/ionization (MALDI) offers potential for fast spatial profiling, but MALDI imaging of tryptic peptides is still challenged by spectral crowding and restricted abilities for MALDI MS/MS identification.

Methods

To address these limitations, we developed pyrene-conjugated 2-pyridinecarboxaldehyde (pyr-2PCA) tags for selective N-terminal labeling and enhanced detection sensitivity. The 2PCA reagent exclusively modifies N-terminal α-amines, not lysine ε-amines, as could be confirmed in MALDI-MS, with concentration-dependent side reactions minimized by dilution. A distinct reporter ion produced by 2PCA-labeled peptides in prm-PASEF (MALDI MS/MS) serves as a unique marker for successful labeling.

Results

The covalent conjugation of 2PCA with a pyrene structure results in the pyr-2PCA tag that enables matrix-free, label-assisted laser desorption ionization mass spectrometry (LALDI-MS) measurements of peptides. We demonstrate that labeling with a pyrene-coupled 2PCA tag (pyr-2PCA) prior to tryptic digestion results in the selective detection of N-terminal peptides in LALDI, with no significant off-target labeling.

Conclusions

This study presents the first presentation and characterization of this novel pyr-2PCA tag, thereby laying the groundwork and demonstrating its future potential for MALDI/LALDI-based in situ spatial N-terminomics to study proteolytic processes.

蛋白水解是一种基本的翻译后修饰。液相色谱-串联质谱（LC-MS /MS）工作流程对于降解分析功能强大，但固有地牺牲了空间信息，这是研究生物系统（如异常细胞外基质重塑和肿瘤微环境改变）的关键方面。基质辅助激光解吸/电离（MALDI）技术提供了快速空间分析的潜力，但MALDI成像仍然受到光谱拥挤和MALDI MS/MS鉴定能力的限制。方法针对这些局限性，我们开发了芘偶联的2-吡啶甲酸（pyr-2PCA）标签，用于选择性n端标记和提高检测灵敏度。正如MALDI-MS所证实的那样，2PCA试剂只修饰n端α-胺，而不是赖氨酸ε-胺，稀释后的浓度依赖性副反应最小化。在prm-PASEF （MALDI MS/MS）中，由2pca标记的肽产生的独特报告离子作为成功标记的独特标记。结果2PCA与芘共价偶联得到pyr-2PCA标签，该标签可用于多肽的无基质、标签辅助激光解吸电离质谱（LALDI-MS）测量。我们证明，在胰蛋白酶消化之前用芘偶联2PCA标记（pyr-2PCA）可以选择性地检测LALDI中的n端肽，而没有明显的脱靶标记。本研究首次提出并表征了这种新型pyr-2PCA标签，从而为基于MALDI/ laldi的原位空间n端组学研究蛋白质水解过程奠定了基础，并展示了其未来的潜力。

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引用次数: 0