Journal of Chromatography A最新文献

Multi–class cyanobacterial toxin analysis using hydrophilic interaction liquid chromatography–mass spectrometry 利用亲水相互作用液相色谱-质谱法分析多类蓝藻毒素。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-14 DOI: 10.1016/j.chroma.2024.465483

Krista M Thomas, Elliott J Wright, Daniel G Beach, Pearse McCarron

Cyanobacteria produce diverse classes of toxins including microcystins, nodularins, anatoxins, cylindrospermopsins and saxitoxins, encompassing a range of chemical properties and mechanisms of toxicity. Comprehensive analysis of these toxins in cyanobacterial, environmental and biological samples generally requires multiple methods of extraction and analysis.

In this work, a method was developed for the major classes of cyanotoxins, which comprised of a three-step liquid-solid extraction method using 75 % CH₃CN with 0.1 % HCOOH and a hydrophilic interaction liquid chromatography (HILIC) elution gradient that provided retention of the less polar microcystins through to the highly polar saxitoxins. Detection was performed by tandem mass spectrometry in selected reaction monitoring mode with positive and negative polarity switching. Identification criteria included matching retention times and product ion ratios with available standards. In-house validation demonstrated good performance of the method including precision ranging from 1.5 (microcystin-LA) to 5.8 (gonyautoxin-2) % RSDs, and detection limits ranging from 0.01 (cylindrospermopsin) to 0.99 (gonyautoxin-3) µg/g in freeze dried material cyanobacteria. Recovery was assessed using spiked non-toxic cyanobacterial samples (Aphanizomenon sp.) and ranged from 83 (neosaxitoxin) to 107 % ([Dha⁷]microcystin-LR).

As a demonstration of application, toxin profiles in cyanobacterial cultures, benthic and planktonic cyanobacteria field samples, and shellfish reference materials were successfully evaluated. The procedure is also amenable for extension to other polar toxin classes including domoic acid and guanitoxin. With increasing reports of cyanobacterial blooms globally, the method represents a powerful quantitative screening tool for measuring cyanotoxins across a broad range of samples.

蓝藻产生的毒素种类繁多，包括微囊藻毒素、球藻毒素、锐毒素、圆柱藻毒素和沙西毒素，其化学性质和毒性机制各不相同。要全面分析蓝藻、环境和生物样本中的这些毒素，通常需要多种提取和分析方法。在这项工作中，针对主要类别的蓝藻毒素开发了一种方法，该方法包括三步液固萃取法，使用 75 % CH3CN 和 0.1 % HCOOH 以及亲水作用液相色谱（HILIC）洗脱梯度，可保留极性较低的微囊藻毒素和极性较高的沙西毒素。检测采用串联质谱法，在选择反应监测模式下进行正负极性切换。鉴定标准包括保留时间和产物离子比是否与现有标准相符。内部验证表明该方法性能良好，在冷冻干燥的蓝藻材料中，精确度从 1.5%（微囊藻毒素-LA）到 5.8%（褐藻毒素-2）不等，检测限从 0.01（圆柱孢藻毒素）到 0.99（褐藻毒素-3）微克/克不等。使用添加的无毒蓝藻样本（Aphanizomenon sp.）对回收率进行了评估，回收率从 83%（新沙蚕毒素）到 107%（[Dha7]微囊藻毒素-LR）不等。作为应用示范，成功评估了蓝藻培养物、底栖和浮游蓝藻现场样本以及贝类参考材料中的毒素概况。该程序还可扩展到其他极性毒素类别，包括多莫西酸和胍基毒素。随着全球有关蓝藻藻华的报道越来越多，该方法是一种强大的定量筛选工具，可用于测量各种样本中的蓝藻毒素。

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引用次数: 0

In situ growth of hierarchical porous covalent organic framework coating for enhanced solid-phase microextraction of phenolic compounds 原位生长分层多孔共价有机框架涂层，用于增强酚类化合物的固相微萃取。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-14 DOI: 10.1016/j.chroma.2024.465519

Weikang Guo, Hui Tao, Haijuan Tao, Qin Shuai, Lijin Huang

Covalent organic frameworks (COFs), when utilized as solid-phase microextraction (SPME) coating materials, exhibit remarkable abilities to concentrate phenols by thousands-fold owing to their vast surface area and exceptional stability. However, the prevalent micropores inherent in COFs can impede rapid mass transfer of target molecules, prolonging SPME extraction times. To addresses this limitation, this work introduces a novel approach by integrating hierarchical porous structures into COFs, leveraging polystyrene microspheres as hard templates during the in situ growth process of the SPME coating. Due to the presence of a hierarchical porous structure derived from the template, the resulting hierarchical porous TpBD coating, termed HP-TpBD, demonstrated enhanced extraction efficiency, accelerated extraction kinetics, and notably shortened extraction times for phenolic compounds. Coupled with gas chromatography-mass spectrometry, a highly sensitive method featuring a low limit of detection (0.20–0.28 ng L^-1), a broad linear range (1.0∼1.0 × 10⁴ ng L^-1), and excellent precision (RSD < 8.5 %) was established. This methodology enables accurate quantitative analysis of phenols in water and soil samples. This work provides valuable insights into developing COF-based SPME coatings for the efficient extraction of volatile contaminants, paving the way for more efficient and sensitive analytical procedures.

共价有机框架（COF）在用作固相微萃取（SPME）涂层材料时，由于其巨大的表面积和优异的稳定性，可将酚类物质浓缩数千倍。然而，COFs 中普遍存在的微孔会阻碍目标分子的快速传质，从而延长 SPME 的萃取时间。为了解决这一局限性，这项工作引入了一种新方法，即在 SPME 涂层的原位生长过程中，利用聚苯乙烯微球作为硬模板，将分层多孔结构集成到 COF 中。由于模板产生了分层多孔结构，由此产生的分层多孔 TpBD 涂层（称为 HP-TpBD）提高了萃取效率，加速了萃取动力学，并显著缩短了酚类化合物的萃取时间。该方法与气相色谱-质谱联用，灵敏度高，检出限低（0.20-0.28 ng L-1），线性范围宽（1.0∼1.0 × 104 ng L-1），精度高（RSD < 8.5 %）。该方法可对水和土壤样品中的酚类物质进行准确的定量分析。这项工作为开发基于 COF 的 SPME 涂层以高效萃取挥发性污染物提供了宝贵的见解，为更高效、更灵敏的分析程序铺平了道路。

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引用次数: 0

Understanding the fundamentals of the on-off retention mechanism of oligonucleotides and their application to high throughput analysis 了解寡核苷酸通断保留机制的基本原理及其在高通量分析中的应用。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-14 DOI: 10.1016/j.chroma.2024.465523

Honorine LARDEUX , Selin BAGCI , Mimi GAO , Wiebke HOLKENJANS , Reinhard PELL , Davy GUILLARME

Ion-pair reversed-phase liquid chromatography (IP-RPLC) is clearly recognized as the gold standard for analyzing therapeutic oligonucleotides (ONs). Recent studies have shown that ONs exhibit an on-off retention behavior in IP-RPLC, meaning that minor changes in acetonitrile (ACN) proportion can significantly impact retention. However, this behavior was initially demonstrated with only a single mobile phase condition. The aim of this study is to gain a deeper understanding of ON elution behavior by measuring the S values (slope of the retention model, log k vs.%ACN) across a broad range of mobile phase conditions. We systematically calculated the S values for both a 20-mer and 100-mer model ON under various conditions, including different IP reagents, IP concentrations, mobile phase pH, column temperatures, and two different buffering acids. We demonstrated that these mobile phase conditions impact the S values in the following order: IP hydrophobicity > IP concentration > column temperature > buffering acid > mobile phase pH. The main explanation for this trend is that mobile phase conditions that reduce the ion-pair retention mechanism (such as low IP hydrophobicity or concentration) will enhance the impact of% ACN on retention, leading to higher S values.

In the second part of the study, this knowledge was used to develop ultra-fast separations for two therapeutic oligonucleotides: a 20-mer antisense oligonucleotide (ASO) without phosphorothioate (PS) modifications and a large single guide RNA (sgRNA) that includes certain PS modifications. The mobile phase conditions were optimized to maximize S values, while preventing the separation of diastereomers. It is important to notice that an S-value of at least 30 is required to benefit from the use of ultra-short columns. This approach allows the successful separation of the main species (ASO and sgRNA) and related impurities in less than one minute using a 5 mm length column.

离子对反相液相色谱（IP-RPLC）被公认为分析治疗性寡核苷酸（ONs）的黄金标准。最近的研究表明，寡核苷酸在 IP-RPLC 中表现出开关保留行为，这意味着乙腈（ACN）比例的微小变化都会对保留产生重大影响。然而，这种行为最初仅在单一流动相条件下得到证实。本研究的目的是通过测量各种流动相条件下的 S 值（保留模型的斜率，log k vs.%ACN ）来深入了解 ON 洗脱行为。我们系统地计算了20聚合物和100聚合物模型ON在不同条件下的S值，包括不同的IP试剂、IP浓度、流动相pH值、色谱柱温度和两种不同的缓冲酸。我们证明，这些流动相条件对 S 值的影响顺序如下：IP 疏水性 > IP 浓度 > 色谱柱温度 > 缓冲酸 > 流动相 pH 值。对这一趋势的主要解释是，降低离子对保留机制的流动相条件（如低 IP 疏水性或浓度）将增强 ACN 对保留的影响，从而导致更高的 S 值。在研究的第二部分，我们利用这一知识开发了两种治疗性寡核苷酸的超快速分离技术：一种是没有硫代磷酸酯（PS）修饰的 20 聚体反义寡核苷酸 (ASO)，另一种是包含某些 PS 修饰的大型单导 RNA (sgRNA)。对流动相条件进行了优化，以最大限度地提高 S 值，同时防止非对映异构体的分离。值得注意的是，要从超短柱的使用中获益，S 值至少要达到 30。使用这种方法，只需使用 5 毫米长的色谱柱，就能在一分钟内成功分离主要物种（ASO 和 sgRNA）和相关杂质。

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引用次数: 0

Designing boron-doped carbon dot-functionalized COFs for fluorescence screening and liquid chromatography tandem mass spectrometry detection of toxins 设计用于荧光筛选和液相色谱串联质谱检测毒素的掺硼碳点功能化 COF。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-12 DOI: 10.1016/j.chroma.2024.465515

Yin-Hai Li , Fei Xu , Wei-Long Zhao , Xiao-Fan Tang , Feng Liu , Chun-Miao Bo

In recent years, mushroom poisoning has been one of the most important factors of food poisoning in China, timely identification of the toxins contained in mushrooms is crucial for the treatment of patients. In this study, boric acid carbon dots (BA-CDs) can undergo specific boron affinity reactions with amatoxins toxins containing o-dihydroxyl groups by means of boric acid groups. Functional covalent organic framework (COF) and BA-CDs were combined to design a adsorbent with boric acid group (COF@VBC@BA-CDs) was designed to meet the requirements of both fluorescent and pretreated materials for amatoxins. The mushrooms and urine samples were rapid screening using fluorescence detection, and then, for positive samples, the target analytes on the COF@VBC@BA-CDs are collected and eluted for next liquid chromatography tandem mass spectrometry (LC-MS/MS) detection. According to the fluorescence characteristics of COF@VBC@BA-CDs, the fluorescence quenching intensity was linearly correlated with the concentration (2–200 μg/L) and the detection limit was 1.2 μg/L. Meanwhile, the detection limit of LC-MS/MS was 0.5 μg/kg for musroom and 0.2 μg/L for urine, as well as the recovery rate was 72.7–110.1%. This noval method meets the methodological requirements and can be used for actual sample analysis.

近年来，蘑菇中毒已成为我国食物中毒的重要因素之一，及时鉴定蘑菇中含有的毒素对患者的治疗至关重要。本研究中，硼酸碳点（BA-CDs）可通过硼酸基团与含有邻二羟基的金针菇毒素发生特异性硼亲和性反应。将功能共价有机框架（COF）和BA-CDs结合起来，设计出一种带有硼酸基团的吸附剂（COF@VBC@BA-CDs），以满足对羊毒素荧光材料和预处理材料的要求。采用荧光检测法对蘑菇和尿液样品进行快速筛查，对阳性样品收集并洗脱 COF@VBC@BA-CDs 上的目标分析物，进行下一步液相色谱串联质谱检测。根据 COF@VBC@BA-CDs 的荧光特性，其荧光淬灭强度与浓度（2-200 μg/L）呈线性关系，检测限为 1.2 μg/L。同时，LC-MS/MS 的检出限为麝香 0.5 μg/kg，尿液 0.2 μg/L，回收率为 72.7-110.1%。该方法符合方法学要求，可用于实际样品分析。

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引用次数: 0

Continuous inertial alignment and isolation of spherical microparticles and nonspherical flagellate microalgae 球形微颗粒和非球形鞭毛微藻的连续惯性排列和分离。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-09 DOI: 10.1016/j.chroma.2024.465509

Xiaoming Chen , Chungang Wu , Jishun Shi , Zhipeng Song , Yingxuan Liu , Bo Han , Zhouyang Zhang , Yong Zhao

Flagellate microalgae play an increasingly significant role in environmental management and biotechnology for valuable bioproducts, excellent photosynthetic capability, and autonomic movement. However, multiple flagellate microalgae practically live together in the ocean and lake areas, and they are susceptible to contamination as a result of improper operations. Enthused by these aspects, we develop a reliable inertial microfluidic method to overcome the influence of flagella movement and non-spherical shape on the alignment and isolation of target flagellate microalgae. Firstly, a computational model incorporating fluid-structure interaction was established to investigate influence of releasing position and shape parameters on the displacement and rotation of non-spherical microalgal cells and numerically studied the processes of shape- and size-based particle separation. Secondly, the movement of different-size particles under diverse flow rates in the channel was explored, and the capability of this method was validated by aligning and separating 10 μm and 20 μm polystyrene particles. Thirdly, this method was applied to align H. pluvialis and isolate Dunaliella salina from the mixed microalgal samples to explore the influence of flow rate on the alignment and isolation of flagellate microalgae. Fourthly, this method was engineered to select 20 μm polystyrene particles from three types of particles and isolate H. pluvialis from the mixture of multiple microalgae species. Finally, we leveraged this approach to realize separation of H. pluvialis and Synedra ulna to explore the performance of this method in shape-based cell separation, and we isolated Euglena from microalgal cell wastes, including dead cells, bacteria, and particles. This method has promising prospects to be a reliable tool to isolate target flagellate microalgae to address problematic issues in environmental monitoring, pharmaceutical synthesis, and chronic wound treatment for the advantage of good adaptability and reliability.

鞭毛纲微藻类因其宝贵的生物产品、出色的光合作用能力和自主运动能力，在环境管理和生物技术领域发挥着越来越重要的作用。然而，在海洋和湖泊地区，多种鞭毛微藻几乎生活在一起，而且由于操作不当，它们很容易受到污染。有鉴于此，我们开发了一种可靠的惯性微流控方法，以克服鞭毛运动和非球形形状对目标鞭毛微藻配准和分离的影响。首先，建立了包含流体与结构相互作用的计算模型，研究了释放位置和形状参数对非球形微藻细胞位移和旋转的影响，并对基于形状和大小的颗粒分离过程进行了数值研究。其次，探讨了不同尺寸颗粒在不同流速下在通道中的运动，并通过排列和分离 10 μm 和 20 μm 聚苯乙烯颗粒验证了该方法的能力。第三，应用该方法对混合微藻样品中的 H. pluvialis 进行配准并分离出 Dunaliella salina，以探索流速对配准和分离鞭毛微藻的影响。第四，我们设计了这种方法，从三种类型的颗粒中选择 20 μm 的聚苯乙烯颗粒，并从多种微藻的混合物中分离出 H. pluvialis。最后，我们利用这种方法实现了褐藻和溃疡藻的分离，探索了这种方法在基于形状的细胞分离中的性能，并从死细胞、细菌和颗粒等微藻细胞废物中分离出了褐藻。该方法具有良好的适应性和可靠性，有望成为分离目标鞭毛微藻的可靠工具，以解决环境监测、药物合成和慢性伤口治疗等方面的问题。

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引用次数: 0

Chromatographic properties of deamidated peptides with Asn-Gly sequences in proteomic bottom-up experiments 蛋白质组自下而上实验中具有 Asn-Gly 序列的脱酰胺肽的色谱特性。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-09 DOI: 10.1016/j.chroma.2024.465513

Quinn Neale , Haley Neustaeter , Vic Spicer , Oleg V. Krokhin

Studies surrounding deamidation have relied on the chromatographic and mass spectrometric differentiation of Asn containing peptides and their isomeric Asp and isoAsp products. The development of mass spectrometry analytical techniques and characterization of isomer specific fragmentation patterns has permitted the investigation of some deamidation species but has struggled to remain effective when applied and on complex samples or in high throughput scenarios. On the other hand, chromatographic separations can provide additional information to facilitate detection of deamidation. In this work the retention characteristics of deamidation products have been reported in reversed-phase separations using formic acid as an ion-pairing modifier. We found three major elution patterns depending on primary and secondary structure of Asn-Gly containing tryptic peptides. Random coil, helical conformations, and N-terminal positioning of Asn usually result in Asn < isoAsp < Asp, isoAsp < Asn < Asp, and Asn < Asp < isoAsp elution order, respectively. These trends, found from the analyses of proteomic samples, were subsequently confirmed via analytical scale UV-HPLC. Additionally, we determined the retention shifts following deamidation for twenty various separation settings used as a first-dimension fractionation for high-throughput proteomic 2D LC-MS/MS analyses.

围绕脱酰胺作用的研究一直依赖于色谱和质谱法来区分含 Asn 的肽及其异构体 Asp 和 isoAsp 产物。质谱分析技术的发展和异构体特异性片段模式的表征使得对某些脱酰胺物种的研究成为可能，但在应用于复杂样品或高通量情况下，质谱分析技术仍难以保持其有效性。另一方面，色谱分离可以提供更多信息，促进脱酰胺的检测。在这项工作中，使用甲酸作为离子配对修饰剂，报告了在反相分离中脱氨产物的保留特征。我们发现，根据含胰蛋白酶肽的一级和二级结构，有三种主要的洗脱模式。Asn的随机线圈、螺旋构象和N端定位通常分别导致Asn < isoAsp < Asp、isoAsp < Asn < Asp和Asn < Asp < isoAsp的洗脱顺序。这些趋势是从蛋白质组样品分析中发现的，随后通过分析规模的 UV-HPLC 得到了证实。此外，我们还确定了 20 种不同分离设置下脱氨后的保留位移，这些分离设置被用作高通量蛋白质组 2D LC-MS/MS 分析的一维分馏。

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引用次数: 0

A systematic approach for estimating colloidal particle adsorption model parameters 估算胶体颗粒吸附模型参数的系统方法。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-09 DOI: 10.1016/j.chroma.2024.465512

Oliver Lorenz-Cristea, Angela Wiebe, Judith Thoma, Maik Veelders, Till Briskot, Simon Kluters, Gang Wang, David Saleh , Federico Rischawy

The estimation of ion-exchange chromatography model parameters is crucial to enable efficient model-assisted biopharmaceutical downstream process development. Model calibration methods can be hindered by model limitations combined with parameter correlations, leading to time-consuming repeated parameter estimations. While Steric Mass Action isotherm estimation methods exist, there is a need for a systematic approach to estimate model parameters for an emerging Colloidal Particle Adsorption (CPA) model proposed by Briskot et al. This study presents a novel strategy that addresses this challenge, offering significant improvements.

Through a parameter sensitivity analysis, we identified key levers for improved CPA parameter estimation, enabling the prediction of elution behavior for low and high load densities in gradient and step elution mode. This analysis also revealed the correlation structure of parameters, allowing the establishment of a minimalized experimental data set for parameter estimation, by using one breakthrough, a high load and three low load density gradient elution experiments. Our workflow leverages a surrogate-assisted global-optimization tool, minimizing computationally expensive function evaluations during parameter fitting. Furthermore, we employed a customized objective function, specifically adapted to the model structure and sensitivity results, to enhance the solver's performance.

Our strategy was tested on three model proteins with molecular weights of approximately 50, 150 and 200 kDa using a strong cation exchange Poros 50 HS resin. Our final approach enabled high throughput CPA model calibration for single components. The resulting CPA models were able to describe non-binding protein-pulses, low and high-loaded gradient elution, break through, as well as isocratic elution experiments.

离子交换色谱模型参数的估算对于实现高效的模型辅助生物制药下游工艺开发至关重要。模型校准方法可能会受到模型限制和参数相关性的阻碍，导致重复参数估计耗时。虽然存在立体质量作用等温线估算方法，但仍需要一种系统方法来估算 Briskot 等人提出的新兴胶体粒子吸附 (CPA) 模型的模型参数。通过参数敏感性分析，我们确定了改进 CPA 参数估计的关键杠杆，从而能够预测梯度和阶跃洗脱模式下低负荷密度和高负荷密度的洗脱行为。这项分析还揭示了参数的相关结构，通过使用一次突破性实验、一次高负荷实验和三次低负荷密度梯度洗脱实验，建立了用于参数估计的最小化实验数据集。我们的工作流程利用了代用辅助全局优化工具，最大限度地减少了参数拟合过程中计算昂贵的函数评估。此外，我们还采用了专门针对模型结构和灵敏度结果而定制的目标函数，以提高求解器的性能。我们使用强阳离子交换 Poros 50 HS 树脂对分子量约为 50、150 和 200 kDa 的三种模型蛋白质进行了测试。我们的最终方法实现了对单一成分的高通量 CPA 模型校准。由此产生的 CPA 模型能够描述非结合蛋白脉冲、低负荷和高负荷梯度洗脱、突破以及等度洗脱实验。

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引用次数: 0

Investigation and characterization of deep eutectic solvent (DES) based stationary phase in gas chromatography 基于深共晶溶剂 (DES) 的气相色谱固定相的研究与表征。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-08 DOI: 10.1016/j.chroma.2024.465511

Amir Mohammad Azadeh, Kourosh Tabar Heydar, Mohammad Hassan Amini

In this study, the preparation of the stationary phase based on the deep eutectic solvent (DES) was investigated along with the measurement of thermodynamic parameters. The physical characterization of the synthesized sample was investigated by Thermogravimetric (TGA), scanning electron microscope (SEM) and Fourier Transform Infrared Spectrometer (FTIR) analyses. Thermodynamic parameters measured to express the potential ability of the constructed stationary phase were McReynolds constants, partition coefficient, Abraham system constants, ∆, activity coefficient at infinite dilution (

γ_{i}^{\infty}

), and selectivity (

S_{ij}^{\infty}

). For this purpose, the parameters were investigated in different amounts of stationary phase (SP) loading including 5, 10, 15 and 18 %w/w and also at three temperatures of 40, 70 and 100 °C. The prepared stationary phase with moderate polarity had the potential to separate BTEXs, aromatic compounds and alcohols due to strong interactions caused by hydrogen bonds. It was investigated the improved performance of the stationary phase prepared with carbowax. Finally, the performance of SP-DES stationary phase separation was investigated for different analytes including alkanes, alcohols and aromatic compounds.

本研究对基于深共晶溶剂（DES）的固定相的制备进行了研究，并对热力学参数进行了测量。通过热重（TGA）、扫描电子显微镜（SEM）和傅立叶变换红外光谱仪（FTIR）分析研究了合成样品的物理特性。测量的热力学参数包括麦克雷诺常数、分配系数、亚伯拉罕系统常数 ∆ 、无限稀释时的活性系数 (γi∞) 和选择性 (Sij∞)，以表示所构建的固定相的潜在能力。为此，研究了不同固定相（SP）负载量（包括 5%、10%、15% 和 18%w/w）以及 40、70 和 100 °C 三种温度下的参数。所制备的极性适中的固定相由于氢键的强烈相互作用，具有分离 BTEX、芳香族化合物和醇类的潜力。研究还发现，使用卡波蜡制备的固定相性能更好。最后，研究了 SP-DES 固定相分离不同分析物（包括烷烃、醇类和芳香族化合物）的性能。

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引用次数: 0

Determination of strobilurin fungicides residual in vegetables based on amino modified magnetic graphene oxide solid phase extraction coupled with GC–MS/MS 基于氨基修饰的磁性氧化石墨烯固相萃取-气相色谱-质谱联用技术测定蔬菜中的残留杀菌剂。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-07 DOI: 10.1016/j.chroma.2024.465455

Qinghua Yang , Qianying Qiu , Yilan Shi , Jinxin Wang , Feng Chen , Juan Yang , Weibing Zhang , Zhen Yin

This research investigated the creation and utilization of an amino-functionalized magnetic graphene oxide (Fe₃O₄-NH₂@GO) nanocomposite as a selective sorbent for extracting and identifying strobilurin fungicides (SFs) from vegetable samples. The investigation utilized a method where magnetic solid phase extraction (MSPE) was integrated with Gas chromatography-triple quadrupole mass spectrometer (GC–MS/MS) for analysis. The physicochemical properties of synthesized sorbent were characterized using Scanning electron microscope (SEM) , Fourier infrared transform spectrum (FT-IR) , X Ray Diffraction (XRD), and vibrating sample magnetometry (VSM) techniques. The adsorption performance of Fe₃O₄-NH₂@GO aligned with the pseudo-second-order kinetics, Elovich, and Freundlich isothermal adsorption models, suggesting various interactions with SFs, including π-π interactions, hydrogen bonding, and electrostatic forces. The maximum adsorption capacity of two selected representative SFs (Picoxystrobin and E-metominostrobin) for the adsorbent were 55.85 mg/g and 47.35 mg/g, respectively. Additionally, the amino group exhibited notable potential for SFs adsorption. Parameters for MSPE, such as the quantity of sorbent, adsorption duration, eluent types and volumes, and sample pH, were optimized. Under optimal conditions, the developed method showed satisfactory performance within a range of 10 to 2000 µg L⁻¹ (R² ≥ 0.9969), low detection limits (0.01–0.080 µg/kg), high analyte recovery (80.5–104.3 %), and good precision (RSD ≤ 4.96 %, n = 6). This method is rapid, straightforward, environmentally friendly, demonstrates good sensitivity and providing a new perspectives on developing sorbents for SFs in complex food matrices. Also Fe₃O₄-NH₂@GO as a sorbent firstly used for extraction and enrichment of phenyl fungicides residues in food samples exhibits remarkable promise, thereby contributing to the advancement of pesticides residue determination methodologies.

本研究调查了氨基功能化磁性氧化石墨烯（Fe₃O₄-NH₂@GO）纳米复合材料作为选择性吸附剂在蔬菜样品中提取和鉴定杀菌剂（SFs）中的应用。研究采用磁性固相萃取（MSPE）与气相色谱-三重四极杆质谱仪（GC-MS/MS）相结合的方法进行分析。利用扫描电子显微镜（SEM）、傅立叶红外光谱（FT-IR）、X 射线衍射（XRD）和振动样品磁力计（VSM）技术对合成吸附剂的理化性质进行了表征。Fe₃O₄-NH₂@GO的吸附性能与伪秒阶动力学、Elovich和Freundlich等温吸附模型一致，表明其与SFs之间存在各种相互作用，包括π-π相互作用、氢键和静电力。吸附剂对两种具有代表性的 SFs（啶氧菌酯和 E-甲氨基菌酯）的最大吸附量分别为 55.85 mg/g 和 47.35 mg/g。此外，氨基在吸附 SFs 方面表现出显著的潜力。对 MSPE 的参数进行了优化，如吸附剂用量、吸附持续时间、洗脱剂类型和用量以及样品 pH 值。在最佳条件下，所建立的方法在10-2000 µg L-¹范围内表现出令人满意的性能（R² ≥ 0.9969），检出限低（0.01-0.080 µg/kg），分析物回收率高（80.5-104.3 %），精密度好（RSD ≤ 4.96 %，n = 6）。该方法快速、简便、环保、灵敏度高，为开发复杂食品基质中 SFs 的吸附剂提供了新的视角。此外，Fe₃O₄-NH₂@GO 作为一种吸附剂首次用于食品样品中苯基杀菌剂残留的萃取和富集也显示出其广阔的前景，从而为农药残留测定方法的发展做出了贡献。

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引用次数: 0

Ultra-high pressure slalom chromatography: Application to the characterization of large DNA and RNA samples relevant in cell and gene therapy 超高压回旋色谱法：应用于细胞和基因治疗相关的大量 DNA 和 RNA 样品的表征。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2024-11-07 DOI: 10.1016/j.chroma.2024.465487

Fabrice Gritti

Slalom chromatography (SC), initially co-discovered by Boyes and Kasai in the late 1980s, has recently re-emerged as a breakthrough technique to rapidly analyze DNA samples. With the development of cutting-edge ultra-high pressure liquid chromatography (UHPLC) systems and columns, SC now offers enhanced resolution and sensitivity for analyzing large DNA samples. By revisiting the fundamentals of the SC retention mechanism (non-equilibrium separation mode) and considering the physicochemical properties of DNA biopolymers (contour length, extension under shear flow, relaxation time), we provide analytical chemists with a general strategy and framework for selecting the most relevant applications in the expanding field of cell and gene therapy. We then present proof-of-concept data demonstrating the rapid separation (under 2 min) of plasmid digest samples containing linear double-stranded (ds) DNA macromolecules starting from 2 kbp to 25 kbp, as well as the accurate size determination (

\pm

6%) of linear dsDNAs. Additionally, we show rapid baseline separation and quantification of extensible linear dsDNAs, along with the more rigid plasmid dsDNA (supercoiled/circular/nicked circular). We also quantify dsRNA impurities present in vitro transcription (IVT) media used for producing new mRNA therapeutics and assess dsRNA structural heterogeneity (conformational isomers). These findings aim to support in a near future application chemists in addressing emerging bioanalytical challenges in cell and gene therapy by offering advanced SC columns and methods.

回旋色谱法（SC）最初是由 Boyes 和 Kasai 在 20 世纪 80 年代末共同发现的，最近再次成为快速分析 DNA 样品的突破性技术。随着最先进的超高压液相色谱（UHPLC）系统和色谱柱的发展，SC 现可为分析大量 DNA 样品提供更高的分辨率和灵敏度。通过重新审视 SC 保留机制（非平衡分离模式）的基本原理，并考虑 DNA 生物聚合物的理化特性（轮廓长度、剪切流下的延伸、弛豫时间），我们为分析化学家提供了在不断扩大的细胞和基因治疗领域选择最相关应用的一般策略和框架。然后，我们展示了概念验证数据，这些数据证明了能快速分离（不到 2 分钟）含有线性双链 (ds) DNA 大分子的质粒消化样本（从 2 kbp 到 25 kbp），并能准确测定线性 dsDNA 的大小（±6%）。此外，我们还展示了快速基线分离和量化可延伸线性dsDNA以及刚性较强的质粒dsDNA（超卷曲/环状/挑刺环状）的方法。我们还量化了用于生产新型 mRNA 疗法的体外转录 (IVT) 培养基中的 dsRNA 杂质，并评估了 dsRNA 的结构异质性（构象异构体）。这些发现旨在通过提供先进的 SC 色谱柱和方法，帮助应用化学家在不久的将来应对细胞和基因治疗领域新出现的生物分析挑战。

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引用次数: 0