Rapid Communications in Mass Spectrometry最新文献_第3页

Enhancing Chimeric Fragmentation Spectra Deconvolution Using Direct Infusion–Tandem Mass Spectrometry Across High-Resolution Mass Spectrometric Platforms 在高分辨率质谱平台上使用直接输注串联质谱增强嵌合碎片谱反褶积。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-11-16 DOI: 10.1002/rcm.10170

Arina Ivanova, Wei Tang, Carsten Simon, Kai Dührkop, Sebastian Böcker, Gerd Gleixner

Rationale

Direct infusion mass spectrometry (DI-MS) is a rapid analytical technique widely used in omics research and other fields. However, the complexity of DI-MS spectra frequently leads to co-fragmentation of analytes with similar m/z, resulting in chimeric fragmentation spectra that complicate compound identification. A DI-based tandem mass spectrometric method (DI-MS2), which modulates the intensity of precursors and fragments by the stepwise movement of the quadrupole isolation window, has been shown to successfully deconvolute chimeric fragmentation spectra. Yet, its applicability to different instruments and optimisation has not been evaluated.

Method

We evaluate the performance of DI-MS2 on two high-resolution instruments: a linear ion trap-Orbitrap (LIT-Orbitrap) and a quadrupole-Orbitrap (Q-Orbitrap). We examined the impact of six instrumental settings, including mass resolving power, isolation window width, step size between MS2 scans, number of microscans, collision energy and automatic gain control (AGC) target, on the analysis of isobaric mixtures with varying m/z differences.

Results

The LIT-Orbitrap consistently achieved high-quality chimeric spectra deconvolution with an average similarity score of 0.98 despite unexpected intensity modulation patterns. The Q-Orbitrap provided four times faster measurements but showed more variable results: It achieved a similarity score of 0.96 for isobars with a m/z difference larger than 0.02, but only 0.56 for m/z differences of 0.006.

Conclusions

These findings indicate that the DI-MS2 is a robust and flexible method applicable across different MS platforms, though the Q-Orbitrap may be less suited for highly complex samples with multiple peaks per nominal mass. This highlights the potential of the DI-MS2 for structural elucidation of complex biological mixtures. Additionally, we provide initial setting optimisation guidelines to improve spectra deconvolution and measurement speed.

原理：直接输注质谱（Direct infusion mass spectrometry, DI-MS）是一种广泛应用于组学研究等领域的快速分析技术。然而，由于DI-MS谱的复杂性，往往导致具有相似m/z的分析物共破碎，从而产生嵌合破碎谱，使化合物鉴定复杂化。基于di的串联质谱方法（DI-MS2）通过四极隔离窗口的逐步移动来调节前体和碎片的强度，已被证明可以成功地反褶曲嵌合碎片谱。然而，它对不同仪器和优化的适用性尚未得到评估。方法：在线性离子阱-轨道阱（lite - orbitrap）和四极离子阱-轨道阱（Q-Orbitrap）两种高分辨率仪器上评价DI-MS2的性能。我们研究了六种仪器设置，包括质量分辨率、隔离窗宽度、MS2扫描之间的步长、微扫描次数、碰撞能量和自动增益控制（AGC）目标，对具有不同m/z差的等压混合物分析的影响。结果：尽管存在意想不到的强度调制模式，lit_orbitrap仍能获得高质量的嵌合光谱反褶积，平均相似度得分为0.98。Q-Orbitrap提供了四倍快的测量速度，但显示了更多可变的结果：对于m/z差大于0.02的等压线，它的相似性得分为0.96，但对于m/z差为0.006的等压线，它的相似性得分仅为0.56。结论：这些发现表明，DI-MS2是一种稳健且灵活的方法，适用于不同的质谱平台，尽管Q-Orbitrap可能不太适合具有每标称质量多个峰的高度复杂样品。这突出了DI-MS2在复杂生物混合物结构解析方面的潜力。此外，我们还提供了初始设置优化指南，以提高光谱反褶积和测量速度。

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引用次数: 0

Editorial-In Memory of Jean-François Muller. 社论-纪念让-弗朗索瓦·穆勒。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-11-09 DOI: 10.1002/rcm.10168

Laurence Charles, Frédéric Aubriet

引用次数: 0

Direct Identification and Quantification of Flavonoids and Their Structural Isomers Using Ambient Ionization Tandem Mass Spectrometry 环境电离串联质谱法直接鉴定和定量黄酮类化合物及其结构异构体

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-11-08 DOI: 10.1002/rcm.10169

Yanqiu Wang, Liping Xu, Tiange Gu, Hongli Li, David Da Yong Chen

Rationale

Flavonoids are phenolic compounds with many health-benefiting properties. However, differentiating different types of flavonoids and their isomers is challenging due to their highly similar structures of various subtypes and different numbers and sites of substituents. Timely quality evaluation of flavonoid-based products is currently almost impossible.

Methods

An ambient ionization method of direct analysis in real time (DART) ion source and tandem mass spectrometry (MS) was used to characterize the fine structures of flavonoids. Different flavonoid subtypes and their isomers with varied numbers and sites of substituents were subjected to DART ionization and collision-induced fragmentation MS analysis.

Results

Seven classes of flavonoids, including methoxy-substituted compounds, exhibited distinctive fragmentation pathways such as retro-Diels-Alder reactions, cross-ring cleavages, and neutral losses. Many flavonoid isomers produced diagnostic MS² and MS³ fragments through DART-tandem MS, enabling direct identification of various isomers within mixtures. An identification workflow was developed, culminating in the creation of a computational tool called FlavoFinder, which automatically determines flavonoid aglycone subtypes and their isomeric structures.

Conclusions

The method and the structural elucidation program were successfully used for the qualitative and quantitative analysis of different flavonoid isomers from real samples. The analysis procedure is high-throughput and is capable of characterizing complex flavonoid structures without extensive sample pretreatment and front-end chromatographic separations.

黄酮类化合物是酚类化合物，具有许多有益健康的特性。然而，由于不同类型的类黄酮及其异构体的结构高度相似，取代基的数量和位置也不同，因此区分不同类型的类黄酮及其异构体具有挑战性。目前，对类黄酮类产品进行及时的质量评价几乎是不可能的。方法采用环境电离实时直接分析（DART）离子源法和串联质谱法（MS）对黄酮类化合物的精细结构进行表征。不同的类黄酮亚型及其不同取代基数量和位置的异构体进行DART电离和碰撞诱导碎片质谱分析。结果7类黄酮类化合物，包括甲氧基取代化合物，均表现出不同的裂解途径，如逆diols - alder反应、交叉环裂解和中性损失。许多类黄酮异构体通过dart串联质谱产生诊断MS2和MS3片段，从而可以直接鉴定混合物中的各种异构体。开发了一个鉴定工作流程，最终创建了一个名为FlavoFinder的计算工具，该工具可以自动确定类黄酮苷元亚型及其异构体结构。结论该方法和结构解析程序可用于实际样品中不同类黄酮异构体的定性和定量分析。该分析方法具有高通量，能够表征复杂的类黄酮结构，而无需大量的样品前处理和前端色谱分离。

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引用次数: 0

Integration of Multiple Mass Spectrometry Techniques and Molecular Network Analysis With Bioinformatics Approaches Revealed the Mechanism of Jiuwei Xifeng Granules in the Treatment of Tourette's Syndrome 多重质谱技术和分子网络分析结合生物信息学方法揭示九味西风颗粒治疗抽动秽语综合征的机制。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-30 DOI: 10.1002/rcm.10156

Chenning Zhang, Pengfei Zhao, Limin Xing, Ruiqi Zhu, Zhiyan Liu, Ying Li, Shuang Li, Chun Bao, Mo Sun, Zhenzhong Wang, Wei Xiao

Rationale

Jiuwei Xifeng Granules, a Chinese patent medicine developed based on renowned traditional Chinese medicine, has shown significant clinical efficacy in treating Tourette's syndrome. However, there is limited research on its pharmacological compounds and mechanism of action.

Methods

In this study, high-resolution mass spectrometry (HRMS) was employed to analyze the natural organic small molecules in the herbal ingredients of Jiuwei Xifeng Granules and the drug-derived components in the serum, liver, and brain tissues of mice following long-term administration, as well as the protein components in the animal drugs. Meanwhile, inorganic trace elements are also widely assessed in Jiuwei Xifeng Granules. Molecular network analysis is utilized to further analyze the natural organic small molecules. Finally, network pharmacology and molecular docking techniques were employed to investigate the mechanisms of action of the main active ingredients.

Results

In total, 51 natural organic small molecules were identified in the five herbal medicines of Jiuwei Xifeng Granules, and the main active components included Gastrodin, Gentiopicroside, and Geissoschizine methyl ether. A total of 22 drug-derived components were identified in the blood of mice, 18 components were detected in the liver, and 8 were found in the brain tissue. Molecular network analysis further expanded the range of identified natural small molecule organic compounds. Thirty-six proteins were identified from two animal drugs in Jiuwei Xifeng Granules, and 68 trace elements were identified from two mineral medicines in Jiuwei Xifeng Granules. The results of the systems pharmacology analysis confirm that the pharmacological action pathways include serotonergic synapse, neuroactive ligand-receptor interaction, dopaminergic synapse, and cAMP signaling pathway. The main targets included DRD2, SLC6A4, HTR1A, DRD1, and KCNH2.

Conclusion

This study employed a multi-omics approach to comprehensively analyze the active compounds in Jiuwei Xifeng Granules, providing a chemical foundation for elucidating its mechanism of action in treating Tourette's syndrome.

理由：九味西风颗粒是一种以著名中医为基础研制的中成药，对治疗抽动秽语综合征具有显著的临床疗效。然而，对其药理成分和作用机制的研究有限。方法：采用高分辨率质谱法（HRMS）分析九味西风颗粒中草药成分中的天然有机小分子，长期给药小鼠血清、肝脏、脑组织中的药源性成分，以及动物性药物中的蛋白质成分。同时，九味西风颗粒中无机微量元素也得到了广泛的评价。利用分子网络分析法对天然有机小分子进行进一步分析。最后，利用网络药理学和分子对接技术对其主要活性成分的作用机制进行了研究。结果：九味西风颗粒中共鉴定出51种天然有机小分子，主要有效成分为天麻素、龙胆苦苷、葛缕荆甲醚。在小鼠血液中共鉴定出22种药物源性成分，在肝脏中检测到18种成分，在脑组织中发现8种成分。分子网络分析进一步扩大了天然小分子有机化合物的鉴定范围。从九味西风颗粒中分离出2种动物药物36种蛋白质，从九味西风颗粒中分离出2种矿物药物68种微量元素。系统药理学分析结果证实其药理作用途径包括5 -羟色胺能突触、神经活性配体-受体相互作用、多巴胺能突触和cAMP信号通路。主要靶点包括DRD2、SLC6A4、HTR1A、DRD1和KCNH2。结论：本研究采用多组学方法综合分析九味西风颗粒的活性成分，为阐明其治疗抽动秽语综合征的作用机制提供化学基础。

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引用次数: 0

Differentiation of α2–3- and α2–6-Linked Sialic Acid Linkages Using Esterification/Amidation Reactions and Tandem Mass Spectrometry 用酯化/酰胺化反应和串联质谱技术鉴别α2-3 -和α2 - 6-唾液酸键

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-29 DOI: 10.1002/rcm.10165

Wei-Chien Weng, Hsien-Wei Tseng, Chin-Yu Liang, Hsin-Kai Tseng, Ching-Ching Yu, Chun-Cheng Lin, Chi-Kung Ni

Rational

Among various oligosaccharides, sialylated oligosaccharides play crucial roles in both physiological and pathological processes, and sialic acid residues within these structures are often used as biomarkers. Mass spectrometry is a widely utilized tool for glycan structural analysis. However, sialic acid is prone to elimination during the sample preparation and the activation process in tandem mass spectra, resulting in a loss of information on sialylation. Consequently, the identification of sialic acid linkages presents a significant analytical challenge. One established method for differentiating α2 → 3- and α2 → 6-linked sialic acids involves esterification followed by ammonia treatment. In this process, the carboxyl group of α2 → 3-linked sialic acids is converted into an amide, while α2 → 6-linked sialic acids are transformed into esters, resulting in distinguishable molecular weights. However, partial conversion of α2 → 6-linked sialic acids to amides and α2 → 3-linked sialic acids to esters has been observed, which reduces the reliability of this differentiation approach.

Methods

A simple tandem mass spectrometry, based on the dissociation mechanisms of hexose and N-acetylhexosamine, is proposed to differentiate α2 → 3 and α2 → 6 sialic acid linkages after esterification and amidation.

Results

Various oligosaccharides consisting of sialic acid with α2 → 3 or α2 → 6 linkages were studied. For some oligosaccharides, the amidation for α2 → 3 linked sialic acid and esterification for α2 → 6 linked sialic acid are nearly complete, while partial amidation for α2 → 6 linked sialic acid and partial esterification for α2 → 3 linked sialic acid for other oligosaccharides was found, suggesting differentiation cannot be solely dependent on the amidation and esterification (or molecular weights). We showed that the α2 → 3 and α2 → 6 linked sialic acids after esterification/amidation can be distinguished by a simple tandem mass spectrometry.

Conclusions

The proposed tandem mass spectrometry enables accurate differentiation of α2 → 3 and α2 → 6 sialic acid linkages, even when both of them undergo amidation or esterification.

在各种寡糖中，唾液酸寡糖在生理和病理过程中都起着至关重要的作用，唾液酸残基在这些结构中经常被用作生物标志物。质谱法是一种广泛应用于多糖结构分析的工具。然而，在串联质谱中，唾液酸在样品制备和激活过程中容易被消除，导致唾液化信息的丢失。因此，唾液酸连接的鉴定提出了一个重大的分析挑战。已建立的区分α2→3-和α2→6-链唾液酸的方法包括酯化反应和氨处理。在此过程中，α2→3链唾液酸的羧基转化为酰胺，α2→6链唾液酸的羧基转化为酯，从而产生可区分的分子量。然而，α2→6链唾液酸部分转化为酰胺，α2→3链唾液酸部分转化为酯，这降低了这种分化方法的可靠性。方法基于己糖和n -乙酰己糖胺的解离机制，采用简单的串联质谱法对α2→3键和α2→6键在酯化和酰胺化后进行区分。结果研究了由α2→3或α2→6键的唾液酸组成的各种低聚糖。对于某些低聚糖，α2→3连接唾液酸的酰胺化和α2→6连接唾液酸的酯化反应基本完成，而对于其他低聚糖，α2→6连接唾液酸的部分酰胺化和α2→3连接唾液酸的部分酯化反应基本完成，提示分化不能仅仅依赖于酰胺化和酯化反应（或分子量）。结果表明，经过酯化/酰胺化反应的α2→3和α2→6链唾液酸可以用简单的串联质谱法进行区分。结论所建立的串联质谱法能够准确区分α2→3和α2→6唾液酸键，即使它们都发生了酰胺化或酯化反应。

{"title":"Differentiation of α2–3- and α2–6-Linked Sialic Acid Linkages Using Esterification/Amidation Reactions and Tandem Mass Spectrometry","authors":"Wei-Chien Weng, Hsien-Wei Tseng, Chin-Yu Liang, Hsin-Kai Tseng, Ching-Ching Yu, Chun-Cheng Lin, Chi-Kung Ni","doi":"10.1002/rcm.10165","DOIUrl":"https://doi.org/10.1002/rcm.10165","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Rational</h3>\u0000 \u0000 <p>Among various oligosaccharides, sialylated oligosaccharides play crucial roles in both physiological and pathological processes, and sialic acid residues within these structures are often used as biomarkers. Mass spectrometry is a widely utilized tool for glycan structural analysis. However, sialic acid is prone to elimination during the sample preparation and the activation process in tandem mass spectra, resulting in a loss of information on sialylation. Consequently, the identification of sialic acid linkages presents a significant analytical challenge. One established method for differentiating α2 → 3- and α2 → 6-linked sialic acids involves esterification followed by ammonia treatment. In this process, the carboxyl group of α2 → 3-linked sialic acids is converted into an amide, while α2 → 6-linked sialic acids are transformed into esters, resulting in distinguishable molecular weights. However, partial conversion of α2 → 6-linked sialic acids to amides and α2 → 3-linked sialic acids to esters has been observed, which reduces the reliability of this differentiation approach.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A simple tandem mass spectrometry, based on the dissociation mechanisms of hexose and <i>N</i>-acetylhexosamine, is proposed to differentiate α2 → 3 and α2 → 6 sialic acid linkages after esterification and amidation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Various oligosaccharides consisting of sialic acid with α2 → 3 or α2 → 6 linkages were studied. For some oligosaccharides, the amidation for α2 → 3 linked sialic acid and esterification for α2 → 6 linked sialic acid are nearly complete, while partial amidation for α2 → 6 linked sialic acid and partial esterification for α2 → 3 linked sialic acid for other oligosaccharides was found, suggesting differentiation cannot be solely dependent on the amidation and esterification (or molecular weights). We showed that the α2 → 3 and α2 → 6 linked sialic acids after esterification/amidation can be distinguished by a simple tandem mass spectrometry.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The proposed tandem mass spectrometry enables accurate differentiation of α2 → 3 and α2 → 6 sialic acid linkages, even when both of them undergo amidation or esterification.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"40 3","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145385198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In Vitro Metabolite Profiling of 4,4-Diaminodiphenylmethane Using Human Liver S9 Fractions and LC-Orbitrap HRMS: In Silico Toxicity Studies of Its Metabolites 利用人肝脏S9组分和LC-Orbitrap HRMS分析4,4-二氨基二苯甲烷的体外代谢物：其代谢物的硅毒性研究

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-29 DOI: 10.1002/rcm.10164

Sridhar Chinthakindi, Radhika Kawathe, Leela Sairam Andhela

Rationale

4,4-Diaminodiphenylmethane (MDA) is an aromatic amine classified as a Group 2B carcinogen. MDA is widely used as a key ingredient in industrial and consumer products such as epoxy resins, polyurethanes, and dyes. Given its widespread use and potential for occupational and environmental exposure, understanding its hepatic metabolism is crucial in elucidating the toxicological profiles of MDA in humans. However, a limited number of metabolites have been reported in the literature. To address this gap, we conducted a comprehensive metabolite profiling study to identify and characterize all possible in vitro metabolites of MDA.

Methodology

In vitro hepatic metabolism studies were conducted using pooled human liver S9 fractions to stimulate the metabolism of MDA in humans. An untargeted metabolite profiling approach was employed using an LC-Orbitrap-HRMS. Data processing and identification of metabolites were performed using the Compound Discoverer software. A targeted LC–MS/MS method was employed to find the relative abundances of metabolites at various incubation times. In silico toxicity prediction of MDA-derived metabolites was conducted using the ProTox 3.0 online tool.

Results

A total of twelve MDA-derived metabolites were successfully identified, reflecting both Phase I (hydroxylation, desaturation, oxidative deamination) and Phase II (acetylation, sulfation, glucoside conjugation) biotransformation pathways. Among these, N-acetylated and desaturation intermediate-related metabolites were the most abundant. Based on these findings, an in silico toxicological report for the metabolites was proposed.

Conclusions

The Orbitrap LC-HRMS platform enabled comprehensive identification and characterization of in vitro hepatic metabolic profiles of MDA. The predicted toxicological profiles and the discovery of additional metabolite formation beyond the known acetylation pathway provided new insights that can enhance toxicological risk assessment of MDA.

4,4-二氨基二苯甲烷（MDA）是一种芳香胺，被列为2B类致癌物。丙二醛被广泛用作工业和消费品的关键成分，如环氧树脂、聚氨酯和染料。鉴于丙二醛的广泛使用以及潜在的职业和环境暴露，了解其肝脏代谢对于阐明人体丙二醛的毒理学特征至关重要。然而，文献中报道的代谢物数量有限。为了解决这一差距，我们进行了一项全面的代谢物分析研究，以确定和表征所有可能的MDA体外代谢物。方法体外肝代谢研究采用混合的人肝脏S9组分来刺激人体内MDA的代谢。采用LC-Orbitrap-HRMS进行非靶向代谢物分析。使用Compound Discoverer软件进行数据处理和代谢物鉴定。采用靶向LC-MS /MS方法测定不同孵育时间代谢物的相对丰度。使用ProTox 3.0在线工具进行了mda衍生代谢物的硅毒性预测。结果共鉴定出12个mda衍生代谢物，反映了第一阶段（羟基化、去饱和、氧化脱胺）和第二阶段（乙酰化、磺化、糖苷偶联）的生物转化途径。其中，n -乙酰化和去饱和中间体相关代谢物最为丰富。基于这些发现，提出了代谢物的计算机毒理学报告。结论Orbitrap LC-HRMS平台能够全面鉴定和表征MDA的体外肝脏代谢谱。预测的毒理学特征和发现已知乙酰化途径之外的其他代谢物形成提供了新的见解，可以加强MDA的毒理学风险评估。

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引用次数: 0

Integrative Approach Based on DI-MS and LC–MS/MS Analysis for Comprehensive Characterization of Flavonoid Glycoside Isomers From Onychopetalum periquino (Annonaceae) 基于DI‐MS和LC-MS /MS的综合分析方法对凤仙花黄酮苷异构体的综合表征。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-28 DOI: 10.1002/rcm.10163

Ana Paula A. Castro, Bruna R. de Lima, Rebeca dos S. França, Brenda R. C. Leocadio, Francinaldo A. da Silva-Filho, Carromberth C. Fernandes, Afonso D. L. de Souza, Hector H. F. Koolen, Maria L. B. Pinheiro, Felipe M. A. da Silva

Rationale

Currently, mass spectrometry–based approaches constitute the primary strategy for the dereplication of flavonoid glycosides; however, strategies focused on the dereplication of their isomers are still scarce. On the other hand, although flavonoid glycosides frequently occur in the leaves of the Annonaceae family, some species, such as Onychopetalum periquino, remain understudied on this topic.

Methods

The aqueous extract from the leaves of O. periquino was filtered through celite and eluted with methanol. The methanol fraction was analyzed by DI-MS in data-dependent acquisition (DDA) mode with neutral loss scans for flavonoid glycosides recognition. Finally, a set of presumable flavonoid glycosides isomers was analyzed by LC–MS/MS and dereplicated through manual interpretation of MS/MS spectra.

Results

DI-MS analysis revealed the presence of flavonoid glycoside isomers, primarily derived from apigenin, luteolin, and quercetin, which were duly dereplicated through LC–MS/MS analysis as follows: vitexin (1), isovitexin (2), apigenin 7-O-glucoside (3), luteolin 6-C-glucoside (4), luteolin 8-C-glucoside (5), luteolin 7-O-glucoside (6), isoscoparin (7), chrysoeriol 7-O-glucoside (8), diosmetin 7-O-glucoside (9), apigenin 8-C-neohesperidoside (10), apigenin 6-C-neohesperidoside (11), apigenin 7-O-rutinoside (12), quercetin 3-O-glucoside-7-O-rhamnoside (13), and quercetin 3-O-rutinoside (14).

Conclusion

This study demonstrated the efficacy of the proposed strategy in identifying and differentiating flavonoid glycoside isomers in O. periquino leaves. Therefore, this workflow is an effective approach for the comprehensive characterization of flavonoid glycoside isomers in future dereplication works. Additionally, it showed how structural differences, such as the type of linkage and position of sugars, influence their fragmentation patterns.

理论基础：目前，基于质谱的方法构成了黄酮类苷分离的主要策略；然而，关注其异构体重复的策略仍然很少。另一方面，虽然黄酮类苷经常出现在番荔枝科植物的叶子中，但一些物种，如Onychopetalum periquino，对这一主题的研究仍然不足。方法：采用水柱过滤，甲醇洗脱。甲醇馏分采用数据依赖采集（DDA）模式，用中性损失扫描的DI-MS进行类黄酮苷识别。最后，通过LC-MS/MS分析了一组可能的黄酮类苷异构体，并通过MS/MS谱的人工解释进行了去重复。结果：DI-MS分析显示黄酮类苷异构体的存在，主要来源于芹菜素、木犀草素和槲皮素，并通过LC-MS/MS分析得到如下结果：牡荆素(1)、异牡荆素(2)、芹菜素7- o -葡萄糖苷(3)、木犀草素6- c -葡萄糖苷(4)、木犀草素8- c -葡萄糖苷(5)、木犀草素7- o -葡萄糖苷(6)、异莨菪素(7)、黄叶菊醇7- o -葡萄糖苷(8)、薯蓣皂苷7- o -葡萄糖苷(9)、芹菜素8- c -新橙皮苷（10）、芹菜素6- c -新橙皮苷（11）、芹菜素7- o -芦丁苷（12）、槲皮素3- o -葡萄糖苷-7- o -鼠李糖苷（13）、槲皮素3- o -芦丁苷（14）。结论：本研究验证了该方法在鉴别白桦叶中黄酮类苷异构体方面的有效性。因此，该工作流程为今后的反复制工作提供了全面表征黄酮类苷异构体的有效途径。此外，它还显示了结构差异，如糖的连接类型和位置，是如何影响它们的断裂模式的。

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引用次数: 0

An Efficient Solid Phase Extraction Method for Purification and Analysis of Compound-Specific Plant Sugar Stable Hydrogen Isotope Values 一种高效固相萃取法纯化和分析化合物特异性植物糖稳定氢同位素值。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-28 DOI: 10.1002/rcm.10161

Selina Hugger, Meisha Holloway-Phillips, Ansgar Kahmen, Daniel B. Nelson

Rationale

The stable hydrogen isotope composition (δ²H) of plant compounds can serve as environmental or metabolic proxies, but interpretations are hindered by insufficient mechanistic understanding. This can be improved by analyzing the δ²H values of metabolic intermediates, such as sucrose, which is the direct substrate of cellulose and a main transport sugar. However, current preparation methods for carbohydrates in general and sucrose in particular are not time-efficient.

Methods

We evaluated methods that use acetylation of soluble carbohydrate plant extracts to aid in purification as well as isotopic analysis of plant sugars such as sucrose. Extracts were obtained using either hot water or hot 80% ethanol. Acetylated extracts were then purified using an established liquid–liquid separation (LL) method or a new solid phase extraction (SPE) method that we developed. We also evaluated glucose produced from starch after enzymatic digestion. Method performance was evaluated based on quantified yields and the impact on measured δ²H values.

Results

Acetylated sucrose and starch-derived glucose were sufficiently resolved for gas chromatography in all cases. No isotopic biasing was detected for any method. Acetylated sucrose yields differed among methods, with 80% ethanol resulting in approximately threefold higher extraction yield compared to water, and SPE giving smaller but still sufficient yields compared to LL. Sample throughput was doubled with the SPE method compared to the LL method, which allows for larger batch sizes compared to LL.

Conclusions

We developed an efficient method to analyze compound-specific plant carbohydrate δ²H values using gas chromatography-isotope ratio mass spectrometry (GC-IRMS). This can be applied in experiments aimed at investigating the processes that shape cellulose δ²H values, including deconvoluting the metabolic and hydro-climatic sources of isotopic variation.

理论基础：植物化合物的稳定氢同位素组成（δ2H）可以作为环境或代谢指标，但由于机制理解不足，解释受到阻碍。这可以通过分析代谢中间体的δ2H值来改善，如蔗糖，它是纤维素的直接底物和主要的运输糖。然而，目前碳水化合物的制备方法，特别是蔗糖的制备方法并不具有时间效率。方法：我们评估了使用可溶性碳水化合物植物提取物乙酰化的方法，以帮助纯化以及植物糖（如蔗糖）的同位素分析。用热水或80%的热乙醇提取提取物。乙酰化提取液采用已建立的液-液分离（LL）法或新开发的固相萃取（SPE）法进行纯化。我们还评估了淀粉经酶消化后产生的葡萄糖。方法性能根据量化产率和对测量δ2H值的影响进行评估。结果：乙酰化蔗糖和淀粉衍生的葡萄糖在所有情况下都能被气相色谱充分分解。任何方法均未检测到同位素偏倚。不同方法的乙酰化蔗糖的提取率不同，80%乙醇的提取率大约是水的三倍，而固相萃取的提取率虽然小，但仍然足够。与LL方法相比，SPE方法的样品吞吐量增加了一倍，与LL方法相比，SPE方法允许更大的批量。结论：建立了一种高效的气相色谱-同位素比质谱（GC-IRMS）分析植物碳水化合物δ2H值的方法。这可以应用于旨在研究形成纤维素δ2H值的过程的实验，包括反旋同位素变化的代谢和水文气候来源。

{"title":"An Efficient Solid Phase Extraction Method for Purification and Analysis of Compound-Specific Plant Sugar Stable Hydrogen Isotope Values","authors":"Selina Hugger, Meisha Holloway-Phillips, Ansgar Kahmen, Daniel B. Nelson","doi":"10.1002/rcm.10161","DOIUrl":"10.1002/rcm.10161","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>The stable hydrogen isotope composition (<i>δ</i><sup>2</sup>H) of plant compounds can serve as environmental or metabolic proxies, but interpretations are hindered by insufficient mechanistic understanding. This can be improved by analyzing the <i>δ</i><sup>2</sup>H values of metabolic intermediates, such as sucrose, which is the direct substrate of cellulose and a main transport sugar. However, current preparation methods for carbohydrates in general and sucrose in particular are not time-efficient.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We evaluated methods that use acetylation of soluble carbohydrate plant extracts to aid in purification as well as isotopic analysis of plant sugars such as sucrose. Extracts were obtained using either hot water or hot 80% ethanol. Acetylated extracts were then purified using an established liquid–liquid separation (LL) method or a new solid phase extraction (SPE) method that we developed. We also evaluated glucose produced from starch after enzymatic digestion. Method performance was evaluated based on quantified yields and the impact on measured <i>δ</i><sup>2</sup>H values.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Acetylated sucrose and starch-derived glucose were sufficiently resolved for gas chromatography in all cases. No isotopic biasing was detected for any method. Acetylated sucrose yields differed among methods, with 80% ethanol resulting in approximately threefold higher extraction yield compared to water, and SPE giving smaller but still sufficient yields compared to LL. Sample throughput was doubled with the SPE method compared to the LL method, which allows for larger batch sizes compared to LL.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>We developed an efficient method to analyze compound-specific plant carbohydrate <i>δ</i><sup>2</sup>H values using gas chromatography-isotope ratio mass spectrometry (GC-IRMS). This can be applied in experiments aimed at investigating the processes that shape cellulose <i>δ</i><sup>2</sup>H values, including deconvoluting the metabolic and hydro-climatic sources of isotopic variation.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"40 2","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12560618/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145375430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Examining the Molecular Composition of Sub-10-nm Domains With Nano-Projectile Secondary Ion Mass Spectrometry 用纳米抛射二次离子质谱法研究亚10nm结构域的分子组成。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-26 DOI: 10.1002/rcm.10162

Michael A. Shaw, Blanca Aparicio, Jander Cruz, D. S. Verkhoturov, S. V. Verkhoturov, E. A. Schweikert, Michael J. Eller

Background

The use of extreme ultraviolet (EUV) light allows for sub-10-nm domains to be patterned. However, a limitation to continued progress on decreasing the size of device features is the performance of existing chemically amplified resists (CARs). These materials need to be examined at or below the desired feature sizes; thus, there is a pressing need to perform molecular analysis at scales at or below 10 nm.

Methods

Nano-projectile secondary ion mass spectrometry (NP-SIMS) is a surface analysis technique wherein an analyte surface is bombarded with a sequence of individual gold nano-projectiles separated in time and space. Three novel primary ions were tested as prospective projectiles for performing molecular analysis on sub-10-nm domains and compared to the reference projectile [Au₄₀₀]⁴⁺ (n/q = 100). Each projectile was tested on carbon foils, silicon wafers, and a model EUV CAR.

Results

Based on examination of transmission electron microscopy micrographs of impact craters in carbon foil, all three novel projectiles produced craters less than 10 nm in diameter. Secondary ion yields on test samples scaled linearly with projectile momentum. The basis of NP-SIMS analysis is the equivalency of each projectile impact. We examined the equivalency of projectile impacts by calculating the correlation coefficient (Q) of ions from a uniform surface and found that Q did not deviate from unity (no observed correlation) for the reference projectile and the novel projectile n/q = 50.

Conclusions

The novel n/q 50 projectile is capable of sampling domains smaller than 10 nm. Each impact is equivalent and thus can be applied to examine the molecular homogeneity of a nonideal surface and examine surface domains that deviate by 2–3 standard deviations from the mean. This makes it an attractive probe for examining novel EUV resists that attempt to produce patterned features below the current 20-nm critical dimension.

背景：使用极紫外光（EUV）可以对10纳米以下的畴进行图案化。然而，现有化学放大抗蚀剂（CARs）的性能限制了在减小器件特征尺寸方面的持续进展。这些材料需要在所需的特征尺寸或以下进行检查；因此，迫切需要在10nm或以下的尺度上进行分子分析。方法：纳米抛射二次离子质谱（NP-SIMS）是一种表面分析技术，其中被分析物表面被一系列在时间和空间上分离的单个金纳米抛射物轰击。研究人员测试了三种新型原生离子作为准射体，用于亚10nm区域的分子分析，并与参考射体[Au400]4+ (n/q = 100)进行了比较。每个弹丸都在碳箔、硅晶片和EUV CAR模型上进行了测试。结果：通过对碳箔上撞击坑的透射电镜观察，三种新型弹丸产生的撞击坑直径均小于10 nm。测试样品的二次离子产率与抛射动量成线性比例。NP-SIMS分析的基础是每次弹丸撞击的等效性。我们通过计算均匀表面离子的相关系数(Q)来检验弹丸撞击的等效性，发现参考弹丸和n/ Q = 50的新型弹丸的Q没有偏离单位（没有观察到相关性）。结论：新型的n/ q50射弹能够采样小于10 nm的域。每次冲击都是等效的，因此可以应用于检查非理想表面的分子均匀性，并检查偏离平均值2-3个标准差的表面域。这使得它成为一种有吸引力的探针，用于检查试图产生低于当前20nm临界尺寸的图案特征的新型EUV电阻。

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引用次数: 0

Exchange of Dual Clumped Isotopes of CO2 With the Phosphoric Acid–Water System CO2双团块同位素与磷酸-水体系的交换。

IF 1.7 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2025-10-24 DOI: 10.1002/rcm.10159

Peter K. Swart, Chaojin Lu

Rationale

The reaction of carbonate materials with phosphoric acid is a standard method for extracting CO₂, not only for the subsequent analysis of the ratios of ¹³C/¹²C and ¹⁸O/¹⁶O but also for the ratios of ¹³C¹⁸O¹⁶O/¹²C¹⁶O¹⁶O and ¹²C¹⁸O¹⁸O/¹²C¹⁶O¹⁶O and the calculation of the Δ₄₇ and Δ₄₈ values. For the determination of Δ₄₇ and Δ₄₈ values, possible exchange between the CO₂ and H₂O in the acid is of particular importance as this influences the precision and accuracy of the measurement.

Methods

Stochastic and non-stochastic CO₂ was exposed to “fresh” and “used” phosphoric acid at 90°C. Fresh acid is defined as acid that had not previously reacted with calcium carbonate, whereas “used” acid had previously dissolved a significant amount of carbonate material. The δ¹³C, δ¹⁸O, Δ₄₇, and Δ₄₈ values of the CO₂ were then measured using a Thermo-253 plus using conventional methods, and the rate and the amount of exchange were determined.

Results

When exposed to “fresh” phosphoric acid, ¹³C¹⁸O¹⁶O and ¹²C¹⁸O¹⁸O readily exchanged with the H₂O in the acid, attaining Δ₄₇ and Δ₄₈ values close to those expected as a result of equilibration with H₂O at 90°C. In contrast with “used” acid, the rate of exchange of ¹³C¹⁸O¹⁶O with H₂O in the acid was negligible, whereas the exchange of ¹²C¹⁸O¹⁸O, although reduced compared with “fresh” acid, was still significant.

Conclusions

Such data have implications on the precision and accuracy of Δ₄₇ and Δ₄₈ values in systems in which the acid has been previously used to react in multiple samples, as well as in those in which fresh acid is used for every sample. The use of a sacrificial amount of carbonate prior to the reaction of “real” samples will reduce some of the changes observed in this study.

理论基础：碳酸盐材料与磷酸反应是提取CO2的标准方法，不仅可以用于后续13C/12C和18O/16O的比值分析，还可以用于13C18O16O/12C16O16O和12C18O18O/12C16O16O的比值以及Δ47和Δ48值的计算。对于Δ47和Δ48值的测定，酸中CO2和H2O之间可能的交换是特别重要的，因为这会影响测量的精度和准确性。方法：将随机和非随机CO2暴露于90°C的“新鲜”和“用过的”磷酸中。新鲜酸被定义为之前没有与碳酸钙反应的酸，而“用过的”酸之前已经溶解了大量的碳酸盐物质。然后用常规方法用thermo253 plus测量CO2的δ13C、δ18O、Δ47和Δ48值，并测定CO2的交换速率和交换量。结果：当暴露于“新鲜”磷酸时，13C18O16O和12C18O18O很容易与酸中的H2O交换，得到的Δ47和Δ48值接近于与H2O在90°C下平衡的结果。与“用过”酸相比，13C18O16O在酸中与H2O的交换速率可以忽略不计，而12C18O18O的交换速率虽然与“新鲜”酸相比有所降低，但仍然显著。结论：这些数据对先前使用酸在多个样品中反应的系统以及对每个样品使用新鲜酸的系统中Δ47和Δ48值的精度和准确性有影响。在“真实”样品的反应之前使用一定量的碳酸盐将减少本研究中观察到的一些变化。

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引用次数: 0