Journal of Chromatography A最新文献_第10页

Method development of comprehensive two-dimensional liquid chromatography: A new metric for preliminary column selection 综合二维液相色谱法的发展：一种初步选择色谱柱的新方法。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465593

Monica Tirapelle, Fanyi Duanmu, Dian Ning Chia, Maximilian O. Besenhard, Luca Mazzei, Eva Sorensen

To elevate the separation performance, two-dimensional liquid chromatography (2D-LC) uses two chromatographic columns with different stationary phases to diversify solute interactions with the resin, hence providing a second “dimension” to solute-specific separation. Developing methods for 2D-LC starts therefore with preliminary column selection. Selecting columns that yield (metaphorically) orthogonal dimensions is of utmost importance, but remains challenging. Although several metrics exist to quantify column orthogonality, currently there is no established methodology, and none of the existing methods accounts for the non-homogeneity of peak band broadening across each separation dimension.

In this work, we propose a new approach to select columns a priori. This approach is based on critical resolution distribution statistics and implicitly accounts for local peak crowding and peak band broadening. Furthermore, we assess the importance of preliminary column selection during in-silico method development and multi-objective optimization of comprehensive 2D-LC. The comparison of the multi-objective Pareto fronts revealed that column pairs selected with our approach provide better separation quality and reduce analysis time compared to column selections via the most established metrics in the literature. Our results prove the importance of preliminary column selection for method development and optimization of 2D-LC systems, and they also show that choosing the right orthogonality metric (such as that proposed here) is crucial.

为了提高分离性能，二维液相色谱（2D-LC）使用两根不同固定相的色谱柱来分散溶质与树脂的相互作用，从而为溶质特异性分离提供了第二个“维度”。因此，2D-LC的开发方法从初步的色谱柱选择开始。选择产生（比方说）正交维的列是极其重要的，但仍然具有挑战性。虽然存在一些量化柱正交性的指标，但目前还没有确定的方法，而且现有的方法都没有考虑到每个分离维度上峰带加宽的非均匀性。在这项工作中，我们提出了一种新的方法来选择列先验。该方法基于临界分辨率分布统计，隐含地考虑了局部峰拥挤和峰带展宽。此外，我们评估了初步柱选择在硅方法开发和综合2D-LC的多目标优化中的重要性。多目标帕累托前沿的比较表明，与通过文献中最成熟的指标进行列选择相比，我们的方法选择的列对提供了更好的分离质量并减少了分析时间。我们的结果证明了初步的色谱柱选择对于2D-LC系统的方法开发和优化的重要性，并且它们还表明选择正确的正交度（例如这里提出的）是至关重要的。

{"title":"Method development of comprehensive two-dimensional liquid chromatography: A new metric for preliminary column selection","authors":"Monica Tirapelle, Fanyi Duanmu, Dian Ning Chia, Maximilian O. Besenhard, Luca Mazzei, Eva Sorensen","doi":"10.1016/j.chroma.2024.465593","DOIUrl":"10.1016/j.chroma.2024.465593","url":null,"abstract":"<div><div>To elevate the separation performance, two-dimensional liquid chromatography (2D-LC) uses two chromatographic columns with different stationary phases to diversify solute interactions with the resin, hence providing a second “dimension” to solute-specific separation. Developing methods for 2D-LC starts therefore with preliminary column selection. Selecting columns that yield (metaphorically) orthogonal dimensions is of utmost importance, but remains challenging. Although several metrics exist to quantify column orthogonality, currently there is no established methodology, and none of the existing methods accounts for the non-homogeneity of peak band broadening across each separation dimension.</div><div>In this work, we propose a new approach to select columns a priori. This approach is based on critical resolution distribution statistics and implicitly accounts for local peak crowding and peak band broadening. Furthermore, we assess the importance of preliminary column selection during <em>in-silico</em> method development and multi-objective optimization of comprehensive 2D-LC. The comparison of the multi-objective Pareto fronts revealed that column pairs selected with our approach provide better separation quality and reduce analysis time compared to column selections via the most established metrics in the literature. Our results prove the importance of preliminary column selection for method development and optimization of 2D-LC systems, and they also show that choosing the right orthogonality metric (such as that proposed here) is crucial.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465593"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142875596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Development of a liquid chromatography–mass spectrometry based targeted metabolomics method for discovering diagnostic biomarkers in Kawasaki disease 基于液相色谱-质谱的川崎病诊断性生物标志物靶向代谢组学方法的建立。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465619

Xialin Luo , Jiaqi Tian , Qing Li , Zhonggan Jin , Xiaoyu Fan , Hong Zhang , Haitao Lv , Yi Ju

Kawasaki disease (KD) has emerged as the leading cause of acquired heart disease in children, primarily due to the absence of highly sensitive and specific biomarkers for early and accurate diagnosis. To address this issue, a simple and comprehensive targeted metabolomics method employing ultra high-performance liquid chromatography coupled with Q-TRAP mass spectrometry has been developed to identify new metabolite biomarkers for KD. This method enables the simultaneous quantification of 276 metabolites, covering 60 metabolic pathways, with a particular emphasis on metabolites relevant to KD. The use of nine ISs and commercial quality control samples significantly enhances both accuracy and precision. Through validation and application to serum samples from patients with KD, seventeen differential serum metabolites were identified. The altered metabolites are primarily associated with three functional metabolic pathways: tricarboxylic acid cycle, tryptophan metabolism, and bile acid metabolism, all of which are believed to be involved in the inflammatory and immune responses in KD patients. Ultimately, eight differential metabolites (indole-3-propionic acid, thiamine, indolepyruvic acid, levodopa, l-selenomethionine, isocitric acid, trans-aconitate, and N-acetylasparagine) were identified that could potentially serve as diagnostic biomarkers with the area under the curve values exceeding 0.9. Our targeted metabolomics approach demonstrates applicability in identifying potential metabolite biomarkers for KD and holds great promise in unraveling the intricate pathophysiology of the disease.

川崎病（KD）已成为儿童获得性心脏病的主要原因，主要是由于缺乏用于早期和准确诊断的高灵敏度和特异性生物标志物。为了解决这一问题，我们开发了一种简单而全面的靶向代谢组学方法，采用超高效液相色谱- Q-TRAP质谱联用技术来鉴定KD的新代谢物生物标志物。该方法能够同时定量276种代谢物，涵盖60种代谢途径，特别强调与KD相关的代谢物。使用9个ISs和商业质量控制样品显着提高了准确性和精密度。通过对KD患者血清样本的验证和应用，鉴定出17种不同的血清代谢物。代谢产物的改变主要与三羧酸循环、色氨酸代谢和胆汁酸代谢这三条功能性代谢途径有关，这些代谢途径被认为与KD患者的炎症和免疫反应有关。最终，确定了8种差异代谢物（吲哚-3-丙酸、硫胺素、吲哚乙酸、左旋多巴、l-硒代蛋氨酸、异柠檬酸、反式乌头酸和n -乙酰天冬氨酸），这些代谢物的曲线下面积超过0.9，有可能作为诊断性生物标志物。我们的靶向代谢组学方法证明了识别KD潜在代谢物生物标志物的适用性，并在揭示该疾病复杂的病理生理方面具有很大的希望。

{"title":"Development of a liquid chromatography–mass spectrometry based targeted metabolomics method for discovering diagnostic biomarkers in Kawasaki disease","authors":"Xialin Luo , Jiaqi Tian , Qing Li , Zhonggan Jin , Xiaoyu Fan , Hong Zhang , Haitao Lv , Yi Ju","doi":"10.1016/j.chroma.2024.465619","DOIUrl":"10.1016/j.chroma.2024.465619","url":null,"abstract":"<div><div>Kawasaki disease (KD) has emerged as the leading cause of acquired heart disease in children, primarily due to the absence of highly sensitive and specific biomarkers for early and accurate diagnosis. To address this issue, a simple and comprehensive targeted metabolomics method employing ultra high-performance liquid chromatography coupled with Q-TRAP mass spectrometry has been developed to identify new metabolite biomarkers for KD. This method enables the simultaneous quantification of 276 metabolites, covering 60 metabolic pathways, with a particular emphasis on metabolites relevant to KD. The use of nine ISs and commercial quality control samples significantly enhances both accuracy and precision. Through validation and application to serum samples from patients with KD, seventeen differential serum metabolites were identified. The altered metabolites are primarily associated with three functional metabolic pathways: tricarboxylic acid cycle, tryptophan metabolism, and bile acid metabolism, all of which are believed to be involved in the inflammatory and immune responses in KD patients. Ultimately, eight differential metabolites (indole-3-propionic acid, thiamine, indolepyruvic acid, levodopa, <span>l</span>-selenomethionine, isocitric acid, trans-aconitate, and N-acetylasparagine) were identified that could potentially serve as diagnostic biomarkers with the area under the curve values exceeding 0.9. Our targeted metabolomics approach demonstrates applicability in identifying potential metabolite biomarkers for KD and holds great promise in unraveling the intricate pathophysiology of the disease.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465619"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142902506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Advancing characterization of pyrolysis products: Comprehensive gas chromatography methods for analytes in gas and liquid phases 推进热解产物的表征：气相和液相分析物的综合气相色谱法。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465641

Bernardo A. Souto, Bingxin Hai, Justice Asomaning, David C. Bressler

The development of biofuel technologies depends on the accurate identification and quantification of products from the conversion processes. Given the complexity of the renewable resources, the availability of biomass, and the versatility of conversion methods, there is a need for characterization methods that provide rapid and reliable analysis for various products coming from different conversion processes with minimal sample preparation. This study develops and validates gas chromatography methods that use multiple detectors to analyze pyrolytic compounds in both gas and liquid phases efficiently in a single, rapid run. For gas products, flame ionization and thermal conductivity detectors were used for identification and quantification of organic and inorganic gases, with volume and mass concentrations showing <1 % error between calculated and referenced values. A method for analyzing liquid product containing hydrocarbons and fatty acids was developed without derivatization, using a simultaneous mass spectrometer for identification and a flame ionization detector for quantification. Validation against established methods demonstrated comparable results, utilizing less time. While the simultaneous dual detector feature provides a simple and rapid analytical tool for pyrolysis products, these methods are also adaptable to non-simultaneous instrument setups and are broadly applicable to any general pyrolysis products containing hydrocarbons and oxygenates, making them useful for improving the development of biofuel production technologies.

生物燃料技术的发展取决于对转化过程中产物的准确识别和量化。考虑到可再生资源的复杂性、生物质的可获得性和转化方法的多功能性，需要一种表征方法，以最少的样品制备为来自不同转化过程的各种产品提供快速可靠的分析。本研究开发并验证了气相色谱方法，该方法使用多个检测器在一次快速运行中有效地分析气相和液相中的热解化合物。对于气体产品，使用火焰电离和导热探测器对有机和无机气体进行鉴定和定量，并显示体积和质量浓度

{"title":"Advancing characterization of pyrolysis products: Comprehensive gas chromatography methods for analytes in gas and liquid phases","authors":"Bernardo A. Souto, Bingxin Hai, Justice Asomaning, David C. Bressler","doi":"10.1016/j.chroma.2024.465641","DOIUrl":"10.1016/j.chroma.2024.465641","url":null,"abstract":"<div><div>The development of biofuel technologies depends on the accurate identification and quantification of products from the conversion processes. Given the complexity of the renewable resources, the availability of biomass, and the versatility of conversion methods, there is a need for characterization methods that provide rapid and reliable analysis for various products coming from different conversion processes with minimal sample preparation. This study develops and validates gas chromatography methods that use multiple detectors to analyze pyrolytic compounds in both gas and liquid phases efficiently in a single, rapid run. For gas products, flame ionization and thermal conductivity detectors were used for identification and quantification of organic and inorganic gases, with volume and mass concentrations showing <1 % error between calculated and referenced values. A method for analyzing liquid product containing hydrocarbons and fatty acids was developed without derivatization, using a simultaneous mass spectrometer for identification and a flame ionization detector for quantification. Validation against established methods demonstrated comparable results, utilizing less time. While the simultaneous dual detector feature provides a simple and rapid analytical tool for pyrolysis products, these methods are also adaptable to non-simultaneous instrument setups and are broadly applicable to any general pyrolysis products containing hydrocarbons and oxygenates, making them useful for improving the development of biofuel production technologies.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465641"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142906326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Probabilistic pressure-flow operating space for chromatographic resins using mechanistic modeling 用机械模型建立色谱树脂的概率压力流操作空间。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465601

Chris A. Gerberich, Chaoying Ding, Lee Bink, André C. Dumetz

Pressure drop across chromatography columns at constant bed height is affected by column diameter and can be difficult to predict without large-scale data. Modern resin engineering has decreased the focus on pressure drop due to the widespread use of large beads and rigid base matrices. However, with the recent development of several small-bead resins optimized for bioprocessing entering the market, pressure drop has justified a regain of attention. This work seeks to develop and apply a mechanistic model based on force balances for predicting pressure drop across scales. With this approach, few small-scale experiments can be used for calibration, and pressure-flow data for large-scale packs can then be predicted with minimum at-scale data. The model was first tested using Phenyl Sepharose 6 FF by calibrating at small scale and then validating the model predictions at larger scale. Strong agreement was observed between predicted and experimental results. The model was then calibrated for seven resins and used to determine a probabilistic operating space calculated based on the likelihood of exceeding pressure limits at scale. Safe operating ranges, accounting for the inherent variability in column packing and resin manufacturing, were set for bed height, fluid velocity, and fluid viscosity. The power of this approach is that it enables screening of resins for potential pressure-flow issues at scale before beginning process development and can be used to set flow rate ranges as solution viscosity changes throughout a chromatographic sequence.

在固定的床层高度下，色谱柱的压降受柱径的影响，在没有大规模数据的情况下很难预测。现代树脂工程已经减少了对压力降的关注，由于大珠和刚性基矩阵的广泛使用。然而，随着最近几种用于生物加工的小颗粒树脂的开发进入市场，压力降重新引起了人们的关注。这项工作旨在开发和应用基于力平衡的机械模型来预测跨尺度的压降。通过这种方法，很少的小规模实验可以用于校准，然后可以用最小的尺度数据预测大型包的压力-流量数据。首先使用Phenyl Sepharose 6 FF在小尺度上进行校准，然后在大尺度上验证模型预测。预测结果与实验结果非常吻合。然后，该模型针对7种树脂进行了校准，并用于确定基于超出规模压力限制的可能性计算的概率操作空间。考虑到柱填料和树脂制造过程中固有的可变性，根据床层高度、流体速度和流体粘度设定了安全操作范围。这种方法的强大之处在于，它可以在开始工艺开发之前大规模筛选树脂，以解决潜在的压力流动问题，并且可以用于设置整个色谱序列中溶液粘度变化的流速范围。

{"title":"Probabilistic pressure-flow operating space for chromatographic resins using mechanistic modeling","authors":"Chris A. Gerberich, Chaoying Ding, Lee Bink, André C. Dumetz","doi":"10.1016/j.chroma.2024.465601","DOIUrl":"10.1016/j.chroma.2024.465601","url":null,"abstract":"<div><div>Pressure drop across chromatography columns at constant bed height is affected by column diameter and can be difficult to predict without large-scale data. Modern resin engineering has decreased the focus on pressure drop due to the widespread use of large beads and rigid base matrices. However, with the recent development of several small-bead resins optimized for bioprocessing entering the market, pressure drop has justified a regain of attention. This work seeks to develop and apply a mechanistic model based on force balances for predicting pressure drop across scales. With this approach, few small-scale experiments can be used for calibration, and pressure-flow data for large-scale packs can then be predicted with minimum at-scale data. The model was first tested using Phenyl Sepharose 6 FF by calibrating at small scale and then validating the model predictions at larger scale. Strong agreement was observed between predicted and experimental results. The model was then calibrated for seven resins and used to determine a probabilistic operating space calculated based on the likelihood of exceeding pressure limits at scale. Safe operating ranges, accounting for the inherent variability in column packing and resin manufacturing, were set for bed height, fluid velocity, and fluid viscosity. The power of this approach is that it enables screening of resins for potential pressure-flow issues at scale before beginning process development and can be used to set flow rate ranges as solution viscosity changes throughout a chromatographic sequence.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465601"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142908931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Diatom biosilica for liquid chromatography 硅藻生物二氧化硅液相色谱。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465603

Michał Szumski , Hussam AlSaoud , Izabela Wojtczak , Myroslav Sprynskyy , Renata Gadzała-Kopciuch , Szymon Bocian , Mikołaj Dembek , Marek Potrzebowski , Bogusław Buszewski

This work presents, for the first time, the preparation method and subsequent use of biosilica in column liquid chromatography in reverse-phase mode. Diatom biosilica consists of the siliceous exoskeletons (frustules) of unicellular algae. Controlled cultivation of Pseudostaurosira trainorii diatoms resulted in frustules with an average diameter of approximately 4 µm, sidewall thickness of 1 µm, and a bottom thickness of 110–150 nm. These frustules contained pores (holes) with diameters ranging from 150 to 300 nm. XRD measurements revealed an opal A silica structure, with some lamellar features, and the material was characterized by a surface area of 21.1 m²/g.

The raw material required careful preparation to remove residual organics by heating it. Following this, the surface was modified with octadecyldimethylchlorosilane to create a reverse-phase chromatographic adsorbent. The resulting columns demonstrated good chromatographic performance, with a theoretical plate number (N) of 22,000 plates for alkylbenzenes on a 160 mm long column, and permeability (K_F) of 5.33 × 10⁻¹⁵ m².

The prepared material exhibited lower hydrophobicity compared to the commercially available HALO C18 stationary phase, which can be attributed to its lower surface area and high number of silanol groups. As a result, only partial separation of six polyaromatic hydrocarbons was achieved due to excessive tailing. However, five anti-inflammatory drugs and two veterinary antibiotics were successfully separated.

本文首次介绍了生物二氧化硅的制备方法及其在反相柱液相色谱中的应用。硅藻生物二氧化硅由单细胞藻类的硅质外骨骼组成。通过对照培养，培养出的硅藻平均直径约为4µm，侧壁厚度为1µm，底部厚度为110 ~ 150 nm。这些结晶体包含直径在150到300纳米之间的孔（孔）。XRD测试结果表明，该材料为蛋白石A型二氧化硅结构，具有一定的层状特征，材料的表面积为21.1 m²/g。原料需要仔细制备，通过加热去除残留的有机物。随后，用十八烷基二甲基氯硅烷对表面进行改性，形成反相色谱吸附剂。所得色谱柱表现出良好的色谱性能，在160毫米长的色谱柱上，烷基苯的理论板数(N)为22,000个板，渗透率（KF）为5.33 × 10⁻¹5m²。与市售的HALO C18固定相相比，制备的材料具有较低的疏水性，这可归因于其较低的表面积和较高的硅烷醇基团数量。结果表明，由于尾矿过多，6种多芳烃只能实现部分分离。但成功分离出5种消炎药和2种兽用抗生素。

{"title":"Diatom biosilica for liquid chromatography","authors":"Michał Szumski , Hussam AlSaoud , Izabela Wojtczak , Myroslav Sprynskyy , Renata Gadzała-Kopciuch , Szymon Bocian , Mikołaj Dembek , Marek Potrzebowski , Bogusław Buszewski","doi":"10.1016/j.chroma.2024.465603","DOIUrl":"10.1016/j.chroma.2024.465603","url":null,"abstract":"<div><div>This work presents, for the first time, the preparation method and subsequent use of biosilica in column liquid chromatography in reverse-phase mode. Diatom biosilica consists of the siliceous exoskeletons (frustules) of unicellular algae. Controlled cultivation of Pseudostaurosira trainorii diatoms resulted in frustules with an average diameter of approximately 4 µm, sidewall thickness of 1 µm, and a bottom thickness of 110–150 nm. These frustules contained pores (holes) with diameters ranging from 150 to 300 nm. XRD measurements revealed an opal A silica structure, with some lamellar features, and the material was characterized by a surface area of 21.1 m²/g.</div><div>The raw material required careful preparation to remove residual organics by heating it. Following this, the surface was modified with octadecyldimethylchlorosilane to create a reverse-phase chromatographic adsorbent. The resulting columns demonstrated good chromatographic performance, with a theoretical plate number (N) of 22,000 plates for alkylbenzenes on a 160 mm long column, and permeability (K<sub>F</sub>) of 5.33 × 10⁻¹⁵ m².</div><div>The prepared material exhibited lower hydrophobicity compared to the commercially available HALO C18 stationary phase, which can be attributed to its lower surface area and high number of silanol groups. As a result, only partial separation of six polyaromatic hydrocarbons was achieved due to excessive tailing. However, five anti-inflammatory drugs and two veterinary antibiotics were successfully separated.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465603"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142870965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Methylcellulose enhances resolution in gravitational field-flow fractionation: Going beyond viscosity 甲基纤维素提高重力场流分馏的分辨率：超越粘度。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465614

R. Danusso , G. Iamoni , A. Moles , G. Casagrande , L. Possenti , I. Cetin , D. Lattuada

Gravitational Field-Flow Fractionation (GrFFF) is an elution-based method designed for the separation of particles ranging from a few micrometers up to approximately 100 μm in diameter. Separation occurs over time, with particles being fractionated based on size and other physico-chemical properties. GrFFF takes advantage of gravitational forces acting perpendicularly to a laminar flow in a thin channel. The fluid exhibits a parabolic velocity profile, with the maximum velocity at the center of the channel and zero velocity at the walls. The exit time of particles depends on their equilibrium position relative to the bottom wall. In hyperlayer mode, larger particles elute faster than smaller ones due to their higher velocities within the channel.

This study investigated the effect of adding methylcellulose (MC) to the carrier fluid on the elution behavior - specifically, peak time (t_peak) and resolution (R) - of polystyrene-based (PS) microparticles with sizes of 7, 8, and 10 μm. The results demonstrated that MC not only increases the viscosity of the carrier fluid but also exerts a secondary, predominant effect that improves resolution (R), thereby enhancing the separation of particle populations. This was confirmed by comparing the use of water as the carrier fluid at two different temperatures: 14 °C (high viscosity) and 28 °C (low viscosity). While increasing viscosity by lowering temperature only led to modest reduction in elution time of the fractograms, the addition of MC had a size-dependent effect on the microparticles, significantly improving R without changing other experimental parameters. This suggests the presence of additional phenomena contributing to the improved separation.

In conclusion, the addition of MC to the carrier fluid increases the resolving power of GrFFF, enabling the separation of PS microbeads with a size difference of up to 2 μm. This advancement pushes the boundaries of GrFFF and opens up potential new applications. These studies, conducted on PS microbeads, provide a preliminary basis for future work on cells, which have similar density and size. This could pave the way for improved cell separation in diagnostic applications.

引力场-流分选（GrFFF）是一种基于洗脱的分离方法，用于分离直径从几微米到大约100 μm的颗粒。随着时间的推移，颗粒根据大小和其他物理化学性质进行分选。GrFFF利用重力垂直作用于薄通道中的层流。流体呈抛物线速度分布，最大速度在通道中心，在通道壁上速度为零。粒子的退出时间取决于它们相对于底壁的平衡位置。在超层模式下，较大的粒子比较小的粒子洗脱速度更快，因为它们在通道内的速度更快。本研究考察了在载体液中加入甲基纤维素（MC）对粒径为7、8和10 μm的聚苯乙烯基（PS）微粒洗脱行为的影响，特别是对洗脱峰时间（tpeak）和分辨率(R)的影响。结果表明，MC不仅增加了载体流体的粘度，而且还发挥了次要的、主要的作用，即提高了分辨率(R)，从而增强了颗粒群的分离。通过比较在14°C（高粘度）和28°C（低粘度）两种不同温度下水作为载液的使用情况，证实了这一点。虽然通过降低温度来增加粘度只能适度减少断口图的洗脱时间，但MC的加入对微颗粒具有尺寸依赖效应，在不改变其他实验参数的情况下显著提高了R。这表明存在有助于改进分离的其他现象。综上所述，在载液中加入MC提高了GrFFF的分辨能力，可以分离出尺寸差达2 μm的PS微珠。这一进步推动了GrFFF的界限，并开辟了潜在的新应用。这些研究是在PS微珠上进行的，为未来在具有相似密度和大小的细胞上的研究提供了初步的基础。这可能为改进诊断应用中的细胞分离铺平道路。

{"title":"Methylcellulose enhances resolution in gravitational field-flow fractionation: Going beyond viscosity","authors":"R. Danusso , G. Iamoni , A. Moles , G. Casagrande , L. Possenti , I. Cetin , D. Lattuada","doi":"10.1016/j.chroma.2024.465614","DOIUrl":"10.1016/j.chroma.2024.465614","url":null,"abstract":"<div><div>Gravitational Field-Flow Fractionation (GrFFF) is an elution-based method designed for the separation of particles ranging from a few micrometers up to approximately 100 μm in diameter. Separation occurs over time, with particles being fractionated based on size and other physico-chemical properties. GrFFF takes advantage of gravitational forces acting perpendicularly to a laminar flow in a thin channel. The fluid exhibits a parabolic velocity profile, with the maximum velocity at the center of the channel and zero velocity at the walls. The exit time of particles depends on their equilibrium position relative to the bottom wall. In hyperlayer mode, larger particles elute faster than smaller ones due to their higher velocities within the channel.</div><div>This study investigated the effect of adding methylcellulose (MC) to the carrier fluid on the elution behavior - specifically, peak time (t<sub>peak</sub>) and resolution (R) - of polystyrene-based (PS) microparticles with sizes of 7, 8, and 10 μm. The results demonstrated that MC not only increases the viscosity of the carrier fluid but also exerts a secondary, predominant effect that improves resolution (R), thereby enhancing the separation of particle populations. This was confirmed by comparing the use of water as the carrier fluid at two different temperatures: 14 °C (high viscosity) and 28 °C (low viscosity). While increasing viscosity by lowering temperature only led to modest reduction in elution time of the fractograms, the addition of MC had a size-dependent effect on the microparticles, significantly improving R without changing other experimental parameters. This suggests the presence of additional phenomena contributing to the improved separation.</div><div>In conclusion, the addition of MC to the carrier fluid increases the resolving power of GrFFF, enabling the separation of PS microbeads with a size difference of up to 2 μm. This advancement pushes the boundaries of GrFFF and opens up potential new applications. These studies, conducted on PS microbeads, provide a preliminary basis for future work on cells, which have similar density and size. This could pave the way for improved cell separation in diagnostic applications.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465614"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142890924","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Determination of carbonation rate of precipitated calcium carbonate in carbonation process based on headspace gas chromatography 顶空气相色谱法测定碳酸化过程中沉淀碳酸钙的碳酸化速率。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465616

Hongyue Zhu , Han Zheng , Tinghui Zhang , Qiyu He , Huan Li , Tingliang Yang , Baoshan Yue , Yingchun Luo , Xu Wang , Jiao Xie , Yi Dai

This paper reports a method for determining the carbonation rate (CR) of precipitated calcium carbonate (PCC) during carbonation process based on headspace gas chromatography technique. The method was carried out by simultaneously detecting the signal values of carbon dioxide and oxygen. Then the carbonation rate of precipitated calcium carbonate in the carbonation process can be calculated by the ratio (γ) of carbon dioxide to oxygen based on a new mathematical model. The results showed that the employed method has good precision (the relative standard deviation < 5 %) and accuracy under the optimized equilibrium temperature and time (60 °C and 10 min). The method is simple and has a large measurable range, which can be an reliable tool for determining the carbonation rate in the carbonation process of PCC.

本文报道了一种基于顶空气相色谱法测定沉淀碳酸钙（PCC）碳酸化过程中碳酸化速率（CR）的方法。该方法是通过同时检测二氧化碳和氧气的信号值来实现的。然后根据新的数学模型，用二氧化碳与氧气的比值（γ）来计算碳酸钙在碳化过程中的碳化速率。结果表明，在最佳平衡温度和平衡时间（60℃，10 min）下，该方法具有良好的精密度（相对标准偏差< 5%）和准确度。该方法操作简单，测量范围大，可作为测定PCC碳酸化过程中碳酸化速率的可靠工具。

引用次数: 0

Corrigendum to “Separation and identification of oligonucleotides impurities and degradation products by reversed phase ultra-high performance liquid chromatography using phenyl-bonded stationary phases without ion pairs - a step towards sustainability” [Journal of Chromatography A 1736 (2024) 465380] “使用无离子对苯基键合固定相的反相超高高效液相色谱分离和鉴定寡核苷酸杂质和降解产物-迈向可持续性的一步”[色谱学报]a 1736(2024) 465380]。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465622

Sylwia Studzińska , Szymon Bocian , Luca Rivoira , Ed Faden , Geoff Faden

引用次数: 0

Separation of polar and neutral lipids from mammalian cell lines by high-performance thin-layer chromatography 高效薄层色谱法分离哺乳动物细胞系极性脂质和中性脂质。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465610

Ana Rita Dias Araújo, Océane Buvry, Bruno Antonny, Delphine Debayle

The introduction of high-performance TLC (HPTLC) instrumentation that allows precise control of critical parameters has transformed the technique into an efficient and rapid tool for analyzing various metabolites, namely lipids. Although mass spectrometry (MS) has largely replaced lipid analysis techniques over recent decades due to its comprehensive lipidome profiling capabilities, it typically lacks the rapidity and simplicity of TLC. HPTLC remains advantageous due to its ease of use, simpler data interpretation, and compatibility with complementary techniques. In this study, we established a HPTLC protocol to fractionate both polar and non-polar lipids on a single normal phase plate. Twenty lipid standards were fractionated and the method was successfully applied to whole extracts from six mammalian cell lines. Standards and extracted lipids were applied with an automated sampler, and polar lipids were first fractionated in a 5-step automated gradient elution, followed by the fractionation of neutral lipids in a twin-trough chamber with three different elutions. Plates were automatically sprayed with a modified copper sulfate solution and charred to reveal lipids and obtain the respective chromatograms. LC-MS was used to identify ambiguous bands, thus ensuring the accuracy of lipid identification.

高性能TLC （HPTLC）仪器的引入，可以精确控制关键参数，将该技术转变为分析各种代谢物（即脂质）的高效快速工具。尽管近几十年来，质谱（MS）由于其全面的脂质组分析能力在很大程度上取代了脂质分析技术，但它通常缺乏TLC的快速和简单性。HPTLC仍然具有优势，因为它易于使用，更简单的数据解释，并与补充技术兼容。在这项研究中，我们建立了一个HPTLC协议，以分离极性和非极性脂质在一个单一的正常相板。对20个脂质标准品进行了分离，并成功地将该方法应用于6种哺乳动物细胞系的全提取物。标准品和提取的脂质使用自动进样器，极性脂质首先在5步自动梯度洗脱中分离，然后在双槽腔中用三种不同的洗脱对中性脂质进行分离。用改性硫酸铜溶液自动喷涂板，并烧焦以显示脂质并获得相应的色谱图。采用LC-MS对歧义条带进行识别，保证了脂质鉴定的准确性。

{"title":"Separation of polar and neutral lipids from mammalian cell lines by high-performance thin-layer chromatography","authors":"Ana Rita Dias Araújo, Océane Buvry, Bruno Antonny, Delphine Debayle","doi":"10.1016/j.chroma.2024.465610","DOIUrl":"10.1016/j.chroma.2024.465610","url":null,"abstract":"<div><div>The introduction of high-performance TLC (HPTLC) instrumentation that allows precise control of critical parameters has transformed the technique into an efficient and rapid tool for analyzing various metabolites, namely lipids. Although mass spectrometry (MS) has largely replaced lipid analysis techniques over recent decades due to its comprehensive lipidome profiling capabilities, it typically lacks the rapidity and simplicity of TLC. HPTLC remains advantageous due to its ease of use, simpler data interpretation, and compatibility with complementary techniques. In this study, we established a HPTLC protocol to fractionate both polar and non-polar lipids on a single normal phase plate. Twenty lipid standards were fractionated and the method was successfully applied to whole extracts from six mammalian cell lines. Standards and extracted lipids were applied with an automated sampler, and polar lipids were first fractionated in a 5-step automated gradient elution, followed by the fractionation of neutral lipids in a twin-trough chamber with three different elutions. Plates were automatically sprayed with a modified copper sulfate solution and charred to reveal lipids and obtain the respective chromatograms. LC-MS was used to identify ambiguous bands, thus ensuring the accuracy of lipid identification.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465610"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142997593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Optimization of an analytical method based on the use of zwitterionic- phosphorylcholine -HILIC column for the determination of multiple polar emerging contaminants in reclaimed water 两性离子-磷酸胆碱- hilic色谱柱测定再生水中多极新兴污染物分析方法的优化。

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A

Pub Date : 2025-01-25 DOI: 10.1016/j.chroma.2024.465605

Lorenzo Strani , Barbara Benedetti , Marina Cocchi , Caterina Durante , Guido Perra , Mattia Pietropaolo , Samuele Pellacani , Daniele Tanzilli

The aim of this study was to optimize a Liquid Chromatography Mass Spectrometry (LC-MS) method using a zwitterionic phosphorylcholine HILIC column for the determination of several Persistent and Mobile Organic Contaminants (PMOC) in wastewater samples. An experimental design approach was implemented to both better understand the retention mechanisms of several polar compounds and to find the optimal operating conditions for their detection and quantification. Eleven PMOCs, with logD_pH=7 ranging from -5.27 to 0.24, were considered, including pesticides, artificial sweeteners, pharmaceuticals, and central nervous system stimulants. Key chromatographic variables—such as the initial percentage of the organic mobile phase, temperature, flow rate, gradient time, acid percentage, and the type and concentration of two different salts— were studied to assess their influence on peak areas, retention times and separation efficiency. The results indicated buffer type, flow rate, and initial percentage of organic mobile phase as the most influential factors affecting retention, though the effects were closely related to the chemical and physicochemical properties of the analytes. The optimized instrumental method demonstrated acceptable figures of merit, with recoveries ranging from 49 % to 100 % for all analytes (except taurine, which may require a different experimental preprocessing step). The method also showed satisfactory precision (repeatability of the entire experimental procedure), in terms of Relative Standard Deviation (RSD %), which was <10 % for all analytes. The developed method was successfully applied to the analysis of reclaimed water samples collected in six wastewater treatment plants in two regions of northern Italy. All target ECs were detected and quantified, except for clenbuterol, terbutaline, acesulfame K and 2,4-dichlorophenoxyacetic acid, which were below the detection limit.

本研究的目的是优化使用两性离子磷胆碱HILIC柱的液相色谱-质谱（LC-MS）测定废水样品中几种持久性和流变性有机污染物（PMOC）的方法。为了更好地了解几种极性化合物的保留机制，并找到其检测和定量的最佳操作条件，采用了实验设计方法。考虑了logDpH=7范围为-5.27至0.24的11种PMOCs，包括杀虫剂、人工甜味剂、药物和中枢神经系统兴奋剂。研究了关键的色谱变量，如有机流动相的初始百分比、温度、流速、梯度时间、酸百分比以及两种不同盐的类型和浓度，以评估它们对峰面积、保留时间和分离效率的影响。结果表明，缓冲液类型、流速和有机流动相初始百分比是影响保留率的主要因素，但其影响与分析物的理化性质密切相关。优化的仪器方法显示出可接受的优点数字，所有分析物的回收率在49%到100%之间（牛磺酸除外，可能需要不同的实验预处理步骤）。该方法的精密度（整个实验过程的重复性）令人满意，相对标准偏差（RSD %）为

{"title":"Optimization of an analytical method based on the use of zwitterionic- phosphorylcholine -HILIC column for the determination of multiple polar emerging contaminants in reclaimed water","authors":"Lorenzo Strani , Barbara Benedetti , Marina Cocchi , Caterina Durante , Guido Perra , Mattia Pietropaolo , Samuele Pellacani , Daniele Tanzilli","doi":"10.1016/j.chroma.2024.465605","DOIUrl":"10.1016/j.chroma.2024.465605","url":null,"abstract":"<div><div>The aim of this study was to optimize a Liquid Chromatography Mass Spectrometry (LC-MS) method using a zwitterionic phosphorylcholine HILIC column for the determination of several Persistent and Mobile Organic Contaminants (PMOC) in wastewater samples. An experimental design approach was implemented to both better understand the retention mechanisms of several polar compounds and to find the optimal operating conditions for their detection and quantification. Eleven PMOCs, with logD<sub>pH=7</sub> ranging from -5.27 to 0.24, were considered, including pesticides, artificial sweeteners, pharmaceuticals, and central nervous system stimulants. Key chromatographic variables—such as the initial percentage of the organic mobile phase, temperature, flow rate, gradient time, acid percentage, and the type and concentration of two different salts— were studied to assess their influence on peak areas, retention times and separation efficiency. The results indicated buffer type, flow rate, and initial percentage of organic mobile phase as the most influential factors affecting retention, though the effects were closely related to the chemical and physicochemical properties of the analytes. The optimized instrumental method demonstrated acceptable figures of merit, with recoveries ranging from 49 % to 100 % for all analytes (except taurine, which may require a different experimental preprocessing step). The method also showed satisfactory precision (repeatability of the entire experimental procedure), in terms of Relative Standard Deviation (RSD %), which was <10 % for all analytes. The developed method was successfully applied to the analysis of reclaimed water samples collected in six wastewater treatment plants in two regions of northern Italy. All target ECs were detected and quantified, except for clenbuterol, terbutaline, acesulfame K and 2,4-dichlorophenoxyacetic acid, which were below the detection limit.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1741 ","pages":"Article 465605"},"PeriodicalIF":3.8,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142890983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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