Journal of Chromatography A最新文献_第9页

pH-dependent separation and identification of saponins from Beta vulgaris L. using high-speed countercurrent chromatography and high-resolution mass spectrometry 高速逆流色谱和高分辨率质谱法分离鉴定甜菜皂苷的ph依赖性。

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2025-12-20 DOI: 10.1016/j.chroma.2025.466643

Anna Tekieli , Mariusz Kowalczyk , Sławomir Wybraniec , Aneta Spórna-Kucab

This study investigated the saponins from Beta vulgaris L. cv. Cylindra roots using a combination of semi-preparative high-speed countercurrent chromatography (HSCCC) and ultra high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS/MS). A new solvent system was developed for the HSCCC purification of saponins under varying pH conditions (3, 5, and 7), which significantly affected their separation. The solvent system consisted of 1-butanol, acetone, acetonitrile, and citrate-phosphate buffer (1.0:0.1:0.05:0.8; v/v/v/v).

The pH adjustment in HSCCC noticeably affected separation efficiency, with higher pH values generally leading to faster elution. The separation behavior of saponins depended primarily on the type, number, and arrangement of sugar substituents. In the HSCCC system, compounds bearing more sugar residues tend to be eluted earlier because their greater hydrophilicity enhances interactions with the mobile phase. Despite this overall trend, saponins containing terminal hexose or pentose units were eluted later, indicating that such sugars can significantly extend elution time. Additionally, saponins featuring dioxolane moieties showed a particularly strong pH dependence, with pH adjustments causing reversible shifts in the elution order of isomeric pairs.

Ten predominant saponins, including the highly concentrated betavulgaroside IV and betavulgaroside III, were quantified with high accuracy using UHPLC-HRMS/MS. The total saponin content in Cylindra was found to be 7.2 g/kg dry extract (DE), significantly higher than previously reported for other B. vulgaris cultivars. Additionally, the study identified 47 saponins, including the novel aglycone norhederagenin (m/z 455.31), thus expanding the phytochemical profile of beetroot.

本文对甜菜皂甙进行了研究。采用半制备高速逆流色谱（HSCCC）和超高效液相色谱-高分辨率质谱（UHPLC-HRMS/MS）相结合的方法对圆柱根进行分析。在不同的pH条件（3、5和7）下，开发了一种新的HSCCC纯化皂苷的溶剂体系，该溶剂体系对皂苷的分离有显著影响。溶剂体系由1-丁醇、丙酮、乙腈和柠檬酸盐-磷酸盐缓冲液（1.0:0.1:0.05:0.8;v/v/v/v）组成。HSCCC中pH值的调整对分离效率影响显著，pH值越高，洗脱速度越快。皂苷的分离行为主要取决于糖取代基的类型、数量和排列。在HSCCC体系中，含有更多糖残基的化合物倾向于更早地被洗脱，因为它们更大的亲水性增强了与流动相的相互作用。尽管有这样的总体趋势，但含有末端己糖或戊糖单位的皂苷被洗脱得较晚，这表明这些糖可以显著延长洗脱时间。此外，具有二恶烷基团的皂苷表现出特别强的pH依赖性，pH值的调整会导致同分异构体对的洗脱顺序发生可逆的变化。采用UHPLC-HRMS/MS对10种主要的皂苷进行了定量分析，包括高浓度的倍柳皂苷IV和倍柳皂苷III。总皂苷含量为7.2 g/kg，显著高于其他品种。此外，该研究还鉴定出了47种皂苷，其中包括一种新的苷元北hederagenin (m/z 455.31)，从而扩大了甜菜根的植物化学图谱。

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引用次数: 0

Analysis of mRNA multimerisation (aggregation) using non-denaturing ion-pair reversed-phase liquid chromatography 使用非变性离子对反相液相色谱分析mRNA多聚（聚集）。

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2025-12-24 DOI: 10.1016/j.chroma.2025.466653

Alexandra L.J. Webb , Emma N. Welbourne , Thomas C. Minshull , Kate A. Loveday , Prerna Bora , Zoltán Kis , Gunilla A. Nilsson , Andal Murthy , Eivor Örnskov , Mark J. Dickman

mRNA-based technology has emerged as a new class of medicines with a wide range of applications, including viral vaccines, cancer vaccines, and therapeutics for the treatment of metabolic diseases and cardiovascular conditions. Impurities, including double-stranded RNA (dsRNA), mRNA fragments, and mRNA multimers (aggregates) that result from the manufacturing of mRNA, as well as from subsequent purification, formulation, and storage, can potentially impact the safety and efficacy of mRNA medicines.

mRNA higher-order structures and mRNA multimers (aggregates) can affect translational efficiency and also impact the efficiency of formulation into lipid nanoparticles. mRNA purity is typically analysed using denaturing or partially denaturing methods, precluding the detection of mRNA multimers (aggregates). In this study, we developed and utilised ion-pair reversed-phase HPLC (IP-RP HPLC) under non-denaturing conditions to analyse mRNA multimers. The inclusion of 1 mM Mg²⁺ in the mobile phase stabilises mRNA higher-order structures, RNA:RNA interactions, and the formation of mRNA dimers/multimers, which can be readily separated from the mRNA monomers.

The ability to resolve mRNA monomers from mRNA dimers/multimers was demonstrated for a range of mRNA sequences and lengths. Moreover, we have shown that the relative abundance of mRNA dimers/multimers is concentration dependent. Using the relative percentage of dimer vs concentration of monomer, we were able to determine that the K_d of the interaction between two eGFP mRNA monomers was 82.93 nM. Characterisation and sizing of the mRNA multimers was performed using mass photometry analysis following the purification of mRNA monomer and dimer/multimer peaks using IP-RP HPLC.

Thus, non-denaturing IP-RP demonstrates significant advantages over current approaches for the analysis of mRNA multimers (aggregates). The high-throughput, temperature-dependent profiling of mRNA multimerisation using IP-RP HPLC will enable further comparative studies on the stability of mRNA multimers and provide important insights into potential factors influencing mRNA multimerisation.

基于mrna的技术已成为一类具有广泛应用的新型药物，包括病毒疫苗、癌症疫苗以及用于治疗代谢疾病和心血管疾病的疗法。杂质，包括双链RNA （dsRNA）、mRNA片段和mRNA多聚体（聚集体），这些杂质来自mRNA的制造以及随后的纯化、配方和储存，可能会影响mRNA药物的安全性和有效性。mRNA高阶结构和mRNA多聚体（聚集体）可以影响翻译效率，也影响脂质纳米颗粒配方的效率。mRNA纯度通常使用变性或部分变性方法进行分析，排除了mRNA多聚体（聚集体）的检测。在这项研究中，我们在非变性条件下开发并使用了离子对反相高效液相色谱（IP-RP HPLC）来分析mRNA多聚体。在流动相中加入1mm Mg²⁺可以稳定mRNA的高阶结构、RNA:RNA的相互作用，以及mRNA二聚体/多聚体的形成，这些二聚体/多聚体可以很容易地从mRNA单体中分离出来。从mRNA二聚体/多聚体中分离mRNA单体的能力在mRNA序列和长度范围内得到了证明。此外，我们已经证明mRNA二聚体/多聚体的相对丰度与浓度有关。利用二聚体相对于单体浓度的相对百分比，我们可以确定两个eGFP mRNA单体相互作用的Kd为82.93 nM。在使用IP-RP高效液相色谱纯化mRNA单体和二聚体/多聚体峰后，使用质谱分析对mRNA多聚体进行表征和分级。因此，非变性IP-RP在分析mRNA多聚体（聚合体）方面比目前的方法具有显著的优势。利用IP-RP高效液相色谱对mRNA多聚合进行高通量、温度依赖性分析，将有助于对mRNA多聚合体的稳定性进行进一步比较研究，并为影响mRNA多聚合的潜在因素提供重要见解。

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引用次数: 0

Solid phase extraction based on a coral like structure conjugated microporous polymer for the detection of preservatives in milk and water 基于珊瑚状结构共轭微孔聚合物的固相萃取法检测牛奶和水中的防腐剂

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2025-12-30 DOI: 10.1016/j.chroma.2025.466667

Hai-Long Jiang , Hong-Yan Liu , Xiao-Li Wang , Da-Feng Jiang , Dong-Mei Liu , Jin-Peng Yuan , Xia Wang , Ru-Song Zhao

In this study, a coral like structure conjugated microporous polymer with abundant N/O functional groups was synthesized using a one-step method. The synthesized material was assembled into a solid-phase extraction (SPE) column, and it showed efficient extraction performance for paraben-based preservatives. Under optimized SPE conditions, a high performance liquid chromatographic (HPLC) method with good linear (r≥0.998), low detection limit (0.07-0.19 ng/mL) and high precision (1.8-6.7%, n=6) was developed. The applicability of this method was evaluated by analyzing real samples (milk and water), with spiked recovery rates ranging from 82.8% to 106%, further confirming the accuracy and reliability of the method. The adsorption mechanism was also discussed by simulation calculations, and it was mainly dominated by hydrogen bonding, π-π interactions and van der Waals forces.

本研究采用一步法合成了一种具有丰富N/O官能团的类珊瑚结构共轭微孔聚合物。将合成的材料组装到固相萃取柱中，对羟基苯甲酸酯类防腐剂进行了高效萃取。在优化的SPE条件下，建立了线性好（r≥0.998）、检出限低（0.07 ~ 0.19 ng/mL）、精密度高（1.8 ~ 6.7%,n=6）的高效液相色谱（HPLC）方法。通过对实际样品（牛奶和水）的分析，验证了该方法的适用性，加标回收率在82.8% ~ 106%之间，进一步验证了该方法的准确性和可靠性。通过模拟计算讨论了吸附机理，发现吸附机理主要由氢键、π-π相互作用和范德华力主导。

引用次数: 0

Preparation of a novel C18-sulfobetaine stationary phase based on silica monolith particles for multimodal high performance liquid chromatography 多模态高效液相色谱c18 -磺基甜菜碱固定相的制备

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-08 DOI: 10.1016/j.chroma.2026.466683

Ashraf Ali , Ya Song , Yongxing Hu , Sarah Alharthi , Eman Y. Santali , Wenjie Zhao

Mixed-mode stationary phases enhance separation flexibility for complex samples by integrating multiple retention mechanisms in high performance liquid chromatography (HPLC). In this study, a novel mixed-mode stationary phase was prepared by functionalizing sub-1 µm porous silica monolith particles (pore size ∼ 35 nm) with 4-vinyl-1,3-sulfopropyl pyridinium betaine (VSPB) and octadecyl silane (C18). The stationary phase (SMP-C18-SB) was characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy, transmission electron microscopy, solid-state 13C NMR spectroscopy, Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH) analysis. The resultant stationary phase was packed into stainless-steel column (150×4.6 mm) by slurry packing method with sequential pressure of 80-100 MPA. The SMP-C18-SB column demonstrated excellent separation performance for alkylbenzenes, nucleosides, zwitterionic compounds, peptides, and proteins, facilitated by hydrophobic, hydrogen bonding, and hydrophilic interactions from C18 and SB groups. SMP-C18-SB column exhibited superior separation performance in both reversed-phase and hydrophilic interaction modes. Moreover, the column back pressure was quite low owing to the irregular shape of SMPs. The SMP-C18-SB column also exhibited excellent repeatability, with intra-day relative standard deviations (RSDs) of <0.11% for retention time and <1.02% for peak area (n = 10 each) after nearly 1000 consecutive injection. Owing to its versatile mixed-mode capability, the SMP-C18-SB column could be used for the separation of complex mixtures containing polar, non-polar and zwitteionic compounds.

混合模式固定相在高效液相色谱（HPLC）中集成了多种保留机制，提高了复杂样品的分离灵活性。在这项研究中，用4-乙烯基-1,3-磺基丙基吡啶甜菜碱（VSPB）和十八烷基硅烷（C18）功能化了亚1 μ m多孔硅整体颗粒（孔径约35 nm），制备了一种新型混合模式固定相。采用x射线光电子能谱（XPS）、扫描电镜、透射电镜、固态13C核磁共振谱（NMR）、Brunauer-Emmett-Teller （BET）和Barrett-Joyner-Halenda （BJH）分析对固定相SMP-C18-SB进行了表征。采用浆液填料法将合成的固定相装入不锈钢柱（150×4.6 mm）中，顺序压力为80 ~ 100 MPA。SMP-C18-SB色谱柱对烷基苯、核苷、两性离子化合物、多肽和蛋白质具有优异的分离性能，这得益于C18和SB基团之间的疏水、氢键和亲水性相互作用。SMP-C18-SB柱在反相和亲水两种相互作用模式下均表现出优异的分离性能。此外，由于smp形状不规则，柱背压很低。SMP-C18-SB柱的重复性也很好，连续进样近1000次后，保留时间的日内相对标准偏差（rsd）为0.11%，峰面积的日内相对标准偏差（rsd）为1.02% （n = 10）。SMP-C18-SB色谱柱具有多种混合模式，可用于分离极性、非极性和两性离子化合物的复杂混合物。

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引用次数: 0

Integrated HPLC–UV validation and LC–MS/MS optimization with molecular networking and chemometrics for advanced characterization of chlorogenic acids in Ilex guayusa 结合分子网络和化学计量学的高效液相色谱-紫外验证和LC-MS /MS优化对绿原酸的高级表征

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-22 DOI: 10.1016/j.chroma.2026.466732

Thomas Garzón , Jefferson V. Pastuña-Fasso , Cristian Quiroz-Moreno , Melanie Ochoa-Ocampo , Evencio J. Medina-Villamizar , Nina Espinosa de los Monteros-Silva , Karel Diéguez-Santana , Jessica L. Cooperstone , Zulay Niño-Ruíz , José R. Almeida , Noroska G.S. Mogollón

Chlorogenic acids (caffeoylquinic acid isomers, CQAs) are major phenolic constituents of Ilex guayusa, but their comprehensive profiling in complex plant matrices is hindered by co-elution, overlapping UV spectra, and isomeric similarity in MS/MS. Rather than aiming to fully resolve isomer-specific quantification by MS, here we present an integrated workflow that couples validated HPLC–UV quantification of the major CQA (5-CQA) with an optimized UPLC–MS/MS strategy designed to improve MS1 peak integrity and expand MS/MS coverage for higher-confidence structural annotation. The HPLC–UV method showed excellent performance for targeted quantification of 5-CQA, including strong linearity (r² = 0.998), selectivity, sensitivity (LOQ = 0.25 mg/L), precision, and recovery. For LC–MS/MS, FastDDA acquisition (top-5 vs. top-15 precursors) revealed the expected trade-off between fragmentation depth and MS1 peak quality; however, post-acquisition raw-data merging restored MS1 fidelity and increased the number of detected features by 43%, enabling high-confidence annotation rather than quantitative discrimination of 16 metabolites and the propagation of oxidized CQA-related derivatives using feature-based molecular networking. Multivariate analyses (PCA, volcano plots, HCA) indicated that geographic location exerted the strongest influence on the metabolite composition, followed by sunlight exposure and plant age. Overall, the proposed workflow provides a practical framework that integrates robust chromatographic quantification with MS acquisition and data-processing optimization, thereby enhancing structural characterization and biological interpretation, rather than complete isomer-resolved quantification, of chlorogenic-acid-related chemistry across complex plant-derived and natural product matrices

绿原酸（咖啡酰奎宁酸异构体，CQAs）是绿原酸（咖啡酰奎宁酸异构体）的主要酚类成分，但它们在复杂植物基质中的全面分析受到共洗脱、重叠紫外光谱和质谱/质谱异构体相似性的阻碍。本文提出了一个集成的工作流程，通过优化的UPLC-MS /MS策略，对主要CQA （5-CQA）的HPLC-UV定量进行验证，旨在提高MS1峰的完整性，扩大MS/MS覆盖范围，以获得更高可信度的结构注释。该方法具有良好的线性（r²= 0.998）、选择性、灵敏度（LOQ = 0.25 mg/L）、精密度和回收率。对于LC-MS /MS， FastDDA采集（前5 vs前15前体）揭示了碎片深度和MS1峰质量之间的预期权衡；然而，采集后的原始数据合并恢复了MS1保真度，并将检测到的特征数量增加了43%，从而实现了16种代谢物的高置信度注释，而不是定量区分，并利用基于特征的分子网络传播氧化cqa相关衍生物。多因素分析（PCA、火山图、HCA）表明，地理位置对代谢物组成的影响最大，其次是日照和植物年龄。总的来说，提出的工作流程提供了一个实用的框架，将强大的色谱定量与质谱采集和数据处理优化相结合，从而增强了结构表征和生物学解释，而不是在复杂的植物源性和天然产物基质中对绿原酸相关化学进行完全的异构分辨定量

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引用次数: 0

Molecular dynamics simulations elucidate the structural determinants of size-exclusion chromatography behavior in dimeric G-quadruplex-RHAU 分子动力学模拟阐明了二聚体g -四聚体- rhau中尺寸排除色谱行为的结构决定因素

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-12 DOI: 10.1016/j.chroma.2026.466706

Min Zhu , Long-yu Zhu , Lu-yan An , Ju Wang , Jun-qin Qiao , Han-yue Yang , Wei-juan Zheng , Hong-zhen Lian

Research on G-quadruplexes (G4s)-RNA helicase associated with AU-rich element (RHAU) interaction has facilitated G4s-targeted therapeutic development. Despite this progress, the interaction between dimeric G-quadruplexes (d-G4s) and RHAU remain less explored compared to monomeric structures. Developing convenient and visual methods to elucidate the interaction mechanisms between different d-G4s structures and RHAU can provide new insights into binding modes, thus aiding in the design of molecular tools targeting d-G4s. In this study, we combined molecular dynamics (MD) simulations with size-exclusion chromatography (SEC) and isothermal titration calorimetry (ITC) to investigate structural dependencies in d-G4s-RHAU interactions for the first time. Diverse d-G4s included hybrid non-parallel (d-24TTG), intramolecular tandem parallel (dAGRO100, GGA8), interlocking parallel (93del), and intermolecular stacked parallel (T30695, T30177) structures were analyzed. MD simulations revealed that the distinct SEC retention behaviors in d-G4s-RHAU interactions were mainly governed by binding site accessibility, inter-site steric hindrance, and binding free energy gradients, trends supported by ITC measured affinities. Furthermore, energy decomposition analysis identified Glu26 in RHAU as a critical residue contributing to electrostatic interactions. Mutating to Arg26 substantially decreased the binding free energy (-94.05 ± 0.41 kJ/mol), emphasizing its functional importance. Thus, this work demonstrates that MD simulations are indispensable for revealing the causes of experimental phenomena and understanding the mechanisms underlying chromatographic behavior. This combined strategy not only discerns interaction patterns stemming from structural diversity but facilitates the rapid screening of G4s-targeting molecules.

g -四重复合物(G4s)-RNA解旋酶与富au元素（RHAU）相互作用的研究促进了G4s靶向治疗的开发。尽管取得了这些进展，但与单体结构相比，二聚体g -四聚物（d-G4s）和RHAU之间的相互作用仍然较少被探索。开发方便直观的方法来阐明不同d-G4s结构与RHAU之间的相互作用机制，可以为研究结合模式提供新的见解，从而有助于设计针对d-G4s的分子工具。在这项研究中，我们首次将分子动力学（MD）模拟与尺寸排除色谱（SEC）和等温滴定量热法（ITC）相结合，研究了d-G4s-RHAU相互作用的结构依赖性。分析了不同结构的d-G4s，包括杂化非平行（d-24TTG）、分子内串联平行（dAGRO100、GGA8）、互锁平行（93del）和分子间堆叠平行（T30695、T30177）。MD模拟表明，d-G4s-RHAU相互作用中不同的SEC保留行为主要受结合位点可达性、位点间空间位阻和结合自由能梯度的控制，这一趋势得到了ITC测量亲和性的支持。此外，能量分解分析确定了RHAU中的Glu26是参与静电相互作用的关键残基。突变Arg26显著降低了结合自由能（-94.05±0.41 kJ/mol），强调了其功能的重要性。因此，这项工作证明了MD模拟对于揭示实验现象的原因和理解色谱行为的机制是必不可少的。这种组合策略不仅可以识别源于结构多样性的相互作用模式，还可以促进g4s靶向分子的快速筛选。

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引用次数: 0

Enrichment of pyrethroid insecticides using effervescent-assisted ionic liquid functionalized in melamine sponge for pipette tip micro solid-phase extraction 三聚氰胺海绵微固相萃取泡腾辅助离子液体富集拟除虫菊酯类杀虫剂。

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-03 DOI: 10.1016/j.chroma.2026.466674

Dararat Saenkam , Pirom Suwannasom , Jitlada Vichapong

A preconcentration and detection method using ionic liquid for embedded in melamine sponge for pipette tip micro solid-phase extraction of pyrethroid insecticides coupled to HPLC was developed for trace amount of pyrethroids residue detection. The synthesis of ionic liquid was accomplished by using tributylhexadecylphosphonium bromide ([P₄₄₄₁₂]Br) and potassium hexafluorophosphate (KPF₆), after that effervescent process was carried out by adding sodium carbonate into a centrifuge tube. The as-prepared ionic liquid was then embedded into melamine sponge, and used for pipette tip micro solid-phase extraction of insecticides in fruit juice matrices. After impregnation, the morphological and structural properties of the sorbents were characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR), respectively. Under the selected conditions, the method exhibited wide linear range (2 – 100 μg l^-1) with a coefficient for determination (R²) greater than 0.99. The limit of quantification (LOQ) and limit of detection (LOD) were in the range of 2 – 3 μg l^-1 and 0.7 – 1 μg l^-1, respectively. Good reproducibility (RSD < 4.65%, n = 5) were achieved. Moreover, the proposed method provided high enrichment factors (EF) ranging from 36.50 to 45.52 folds. The sorbent was then applied to extract pyrethroids in fruit juice. Accuracy was excellent and acceptable recoveries were attained. The greenness profiles were evaluated utilizing multiple assessment metrics including the Analytical Eco-Scale, AGREE Assessment, and BAGI.

建立了离子液体包埋在三聚氰胺海绵中进行移管尖端微固相萃取拟除虫菊酯类杀虫剂的预浓缩检测方法，并结合高效液相色谱法对痕量拟除虫菊酯类杀虫剂残留进行检测。以三烷基十六烷基溴化磷（[P44412]Br）和六氟磷酸钾（KPF6）为原料，在离心管中加入碳酸钠进行泡腾制得离子液体。将制备好的离子液体包埋在三聚氰胺海绵中，用于吸管尖微固相萃取果汁基质中的杀虫剂。浸渍后，用扫描电镜（SEM）和傅里叶变换红外光谱（FTIR）分别表征了吸附剂的形态和结构特性。在所选择的条件下，该方法具有较宽的线性范围（2 ~ 100 μg -1），测定系数（R2）大于0.99。定量限和检出限分别在2 ~ 3 μg -1和0.7 ~ 1 μg -1范围内。重现性好（RSD < 4.65%, n = 5）。此外，该方法具有较高的富集因子（EF），范围为36.50 ~ 45.52倍。然后应用该吸附剂提取果汁中的拟除虫菊酯。准确度极好，回收率可接受。利用多种评估指标对绿色概况进行评估，包括分析生态尺度、AGREE评估和BAGI。

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引用次数: 0

Advances in desorption electrospray ionization mass spectrometry imaging: Research progress and applications 解吸电喷雾电离质谱成像的研究进展及应用

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-19 DOI: 10.1016/j.chroma.2026.466719

Xujun Zhang , Shan Wang , Bo Sui , Yan Wang , Lingbo Ji , Jianxun Zhang , Ajuan Yu , Wu Fan , Wuduo Zhao

Desorption electrospray ionization mass spectrometry imaging (DESI-MSI) is regarded as a novel molecular imaging approach that integrates DESI source with conventional mass spectrometry to provide spatial information and molecular composition on the sample’s surface. In this review, the principle of DESI-MSI was elucidated, including its desorption process and ionization mechanism. Through the applications in biological tissues and organs, the advantages of DESI-MSI technology were demonstrated in exploring the tissue functional roles and disease mechanisms, including brain, liver, kidney, and lung. Building on these findings, we review DESI-MSI’s unique capability to map both exogenous and endogenous biomolecules, thereby providing insights into spatial distribution of exogenous organisms and endogenous metabolic of transformation pathways and pharmacokinetics. Finally, the recent applications of DESI-MSI in biomedicine, environmental monitoring, and food safety are also reviewed. With the continuous improvement of instruments and analysis methods, DESI-MSI is expected to bring meaningful changes across a growing number of fields, opening up exciting opportunities to support innovative technologies and applied research.

解吸电喷雾电离质谱成像（DESI- msi）是一种新型的分子成像方法，它将DESI源与传统的质谱相结合，提供样品表面的空间信息和分子组成。本文综述了DESI-MSI的原理，包括其解吸过程和电离机理。通过在生物组织器官中的应用，展示了DESI-MSI技术在探索脑、肝、肾、肺等组织功能作用和疾病机制方面的优势。在这些发现的基础上，我们回顾了DESI-MSI绘制外源性和内源性生物分子的独特能力，从而提供了外源性生物的空间分布和内源性代谢转化途径和药代动力学的见解。最后，对DESI-MSI在生物医学、环境监测、食品安全等方面的应用进行了综述。随着仪器和分析方法的不断改进，DESI-MSI有望在越来越多的领域带来有意义的变化，为支持创新技术和应用研究开辟令人兴奋的机会。

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引用次数: 0

Biophysical characterisation of mRNA structure and its impact on integrity analysis by liquid chromatography and capillary gel electrophoresis methods mRNA结构的生物物理特征及其对液相色谱和毛细管凝胶电泳方法完整性分析的影响

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2025-12-17 DOI: 10.1016/j.chroma.2025.466625

Mayada Elgohary , Alexandra L.J. Webb , Joseph C. Ward , Andal Murthy , Jack Hutchinson , James Button , George Thom , Philip Newton , Emmelie Hammarvid , Anna D. Baran , Juhi Patel , Hannah N. Turney , Mark J. Dickman , Sara Trabulo , Eivor Örnskov

Messenger RNA (mRNA) integrity is a critical quality attribute (CQA) commonly assessed using liquid chromatography (LC) and capillary gel electrophoresis (CGE). The LC and CGE methods used for integrity tests are also used for stability studies for establishing product shelf-life for mRNA-based therapeutics. Advances in sequence and structure engineering have demonstrated significant improvements in mRNA translatability and stability. However, the higher order structure (HOS) of mRNA remains poorly understood and it is difficult to correlate experimentally measured structures with their bioinformatically predicted conformations. Previous studies have shown that mRNA HOS can cause artefactual bands or peaks in CGE and LC analyses, leading to anomalous integrity readouts. In this study, we employed a range of biophysical methods to characterise mRNA HOS, aiming to clarify the origins of the additional peaks observed in CGE and LC analyses and to better comprehend the influences of predicted minimum free energy (MFE) and structural features on mRNA integrity, translatability, and stability. Our findings provide new method-related considerations for measuring mRNA integrity as well as insights into the effects of mRNA structure on stability under long term storage and forced degradation conditions.

信使RNA （mRNA）完整性是一项重要的质量属性（CQA），通常使用液相色谱（LC）和毛细管凝胶电泳（CGE）来评估。用于完整性测试的LC和CGE方法也用于稳定性研究，以确定基于mrna的治疗方法的产品保质期。随着序列和结构工程的发展，mRNA的可译性和稳定性得到了显著改善。然而，mRNA的高阶结构（HOS）仍然知之甚少，并且很难将实验测量的结构与其生物信息学预测的构象联系起来。先前的研究表明，mRNA HOS可以在CGE和LC分析中引起人为的波段或峰，导致异常的完整性读数。在这项研究中，我们采用了一系列生物物理方法来表征mRNA HOS，旨在澄清在CGE和LC分析中观察到的额外峰的起源，并更好地理解预测的最小自由能（MFE）和结构特征对mRNA完整性、可译性和稳定性的影响。我们的研究结果为测量mRNA完整性提供了新的方法相关考虑因素，并深入了解了mRNA结构在长期储存和强制降解条件下对稳定性的影响。

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引用次数: 0

Measuring fluorotelomer alcohols by thermal desorption-gas chromatography-tandem mass spectrometry: Interlaboratory study results 热解吸-气相色谱-串联质谱法测定氟端聚物醇：实验室间研究结果

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2026-02-22 Epub Date: 2026-01-21 DOI: 10.1016/j.chroma.2026.466725

Clara M.A. Eichler , Hannah Calder , Bharat Chandramouli , Matthew Curtis , Heidi Hayes , Benjamin Kim , Ruth Marfil-Vega , Cristina Matos Mejías , Laura Miles , Alan Owens , Jack Stuff , Kurt Thaxton , Jochen Vandenberg , Nicola Watson , David Wevill , Jackie A. Whitecavage , Xiaoyu Liu

Fluorotelomer alcohols (FTOHs) are a group of volatile and semi-volatile per- and polyfluoroalkyl substances (PFAS) commonly found in indoor air and contribute to PFAS inhalation exposure. Improving our understanding of the contribution of FTOHs to human exposure to PFAS indoors is of high interest. Consensus standard test methods play an essential role in environmental risk assessment and management. We present the outcome of an international interlaboratory study (ILS) conducted with nine laboratories to evaluate the precision of ASTM International Standard Test Method D8591. The ILS was organized by the United States Environmental Protection Agency. The test method specifies the analysis of four FTOHs (4:2, 6:2, 8:2 and 10:2 FTOH) collected on PFAS-specific thermal desorption tubes by gas chromatography coupled with tandem mass spectrometry. During the ILS, participating laboratories were instructed to use the test method to analyze three samples (A, B, and C), each three times. Each sample contained the target FTOHs at defined concentrations unknown to the laboratories. The results from seven laboratories show that the relative reproducibility standard deviation (RSD_R) of the method ranges from 14% to 26% and the relative repeatability standard deviation (RSD_r) ranges from 4.6% to 11%, with RSDs decreasing with decreasing volatility of the FTOHs. Bias ranged from -13% to 6.0% and was generally larger and negative for less volatile FTOHs. The test method in conjunction with the precision statistics from this ILS will provide a reliable, defendable method that can be used in the context of studying PFAS sources, transport, and human exposure.

氟调聚物醇（FTOHs）是一组挥发性和半挥发性的单氟烷基和多氟烷基物质（PFAS），通常存在于室内空气中，并导致PFAS吸入暴露。提高我们对FTOHs对人类室内PFAS暴露的贡献的理解是非常有意义的。共识标准测试方法在环境风险评价和管理中起着至关重要的作用。我们提出了一项由9个实验室进行的国际实验室间研究（ILS）的结果，以评估ASTM国际标准测试方法D8591的精度。这次活动是由美国环境保护署组织的。采用气相色谱-串联质谱法对pfas专用热脱附管上采集的4种FTOH（4:2、6:2、8:2和10:2）进行分析。在ILS期间，参与实验室被指示使用测试方法分析三种样品（A， B和C），每三次。每个样品都含有实验室未知的特定浓度的目标FTOHs。7个实验室的结果表明，该方法的相对重复性标准偏差（RSDR）为14% ~ 26%，相对重复性标准偏差（RSDR）为4.6% ~ 11%，rsd随FTOHs挥发性的降低而减小。偏倚范围从-13%到6.0%，对于波动性较小的ftoh，偏倚通常较大且为负。该测试方法与该ILS的精确统计数据相结合，将提供一种可靠的、可防御的方法，可用于研究PFAS的来源、运输和人体暴露。

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引用次数: 0