Journal of Chromatography B最新文献_第6页

Naringin-templated magnetic molecularly imprinted polymers for selective quercetin extraction from onion peel 以柚皮苷为模板的磁性分子印迹聚合物用于从洋葱皮中选择性提取槲皮素

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124349

Vinitha Udhayabanu Govindarajan, Vaishnavi Renganathan, Meenakshi Sundaram Muthuraman

{"title":"Naringin-templated magnetic molecularly imprinted polymers for selective quercetin extraction from onion peel","authors":"Vinitha Udhayabanu Govindarajan, Vaishnavi Renganathan, Meenakshi Sundaram Muthuraman","doi":"10.1016/j.jchromb.2024.124349","DOIUrl":"10.1016/j.jchromb.2024.124349","url":null,"abstract":"<div><div>A Magnetic Molecularly Imprinted Polymer (MMIP) was developed using naringin as template molecule, acrylamide as functional monomer and polymerized by ultrasound irradiation for the adsorption of naringin. In an unexpected turn of results, the selectivity study unveiled that the synthesized MMIP exhibited a higher affinity for quercetin over naringin. Given this high selectivity, adsorption isotherm and kinetic studies were conducted for both quercetin and naringin. The adsorption isotherm indicated multilayer adsorption of the adsorbate on the adsorbent. The kinetic study showed better agreement with the pseudo-second-order kinetic model. The maximum adsorption capacity of 7.2 mg/g was achieved for quercetin at 50 mg/L and 4.9 mg/g was attained for naringin at the same concentration. Furthermore, quercetin quantification was performed by coupling MMIP with HPLC-UV, with method validation revealing the limits of detection (LOD) and quantification (LOQ) for quercetin. Additionally, agro-industrial waste onion peel, enriched with phenolic compounds such as quercetin, was subjected to solid-phase extraction using MMIP for the purification of quercetin.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124349"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142586150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Extension of impurity profiling on eltrombopag olamine to in-silico predictions: An effort to exploit correlated forced degradation products and known drug-related substances in drug discovery 将艾曲波帕乙醇胺上的杂质分析扩展到实验室预测：在药物发现中利用相关强制降解产物和已知药物相关物质的努力。

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124367

Saurabh B. Ganorkar , Preeti S. Bobade , Rakesh C. Prabhu , Deepak K. Lokwani , Ranajit N. Shinde , Darshan R. Telange , Atul A. Shirkhedkar , Yvan Vander Heyden

{"title":"Extension of impurity profiling on eltrombopag olamine to in-silico predictions: An effort to exploit correlated forced degradation products and known drug-related substances in drug discovery","authors":"Saurabh B. Ganorkar , Preeti S. Bobade , Rakesh C. Prabhu , Deepak K. Lokwani , Ranajit N. Shinde , Darshan R. Telange , Atul A. Shirkhedkar , Yvan Vander Heyden","doi":"10.1016/j.jchromb.2024.124367","DOIUrl":"10.1016/j.jchromb.2024.124367","url":null,"abstract":"<div><div>The recent pandemic has highlighted the impact of diseases on global health and the economy. The rapid discovery of new hit molecules remains a tough challenge. Pharmaceutical impurity profiling can be linked to drug discovery through the identification of new hits from compounds identified during the analytical profiling. The present study demonstrates this linkage through the extension of the impurity (forced degradation) profiling of eltrombopag (ELT) olamine, a thrombopoietin (TPO) receptor agonist. The drug was exposed to standard degradation and the degradation products were primarily resolved and identified by UPLC-ESI-MS. This led to the identification of five forced degradation products (FDP). Thirty-three other known related substances (RS) of ELT, identified in the literature, were also considered. Molecular similarity checks were performed using Tanimoto/Jaccard's similarity searches. A set of structurally and topologically similar molecules, including ELT and 15 RS, was established and subjected to in-silico toxicity-, absorption-, distribution-, metabolism-, and elimination (ADME) predictions. The RS, predicted with similar or lower toxicity than ELT and a comparable ADME profile, were subjected to molecular docking to trace changes in TPO receptor affinity. The results indicated that five RS had a high Jaccard’s similarity with ELT and higher or comparable docking scores. These compounds, along with few other impurities were predicted to have lower toxicity, better or comparable absorption, distribution, metabolism, and also a better excretion profile than ELT. This justifies their entry as potential novel TPO receptor agonists in drug discovery.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124367"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142638171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Heteroatom cobalt-based metal-organic framework and reduced graphene oxide nanocomposite for dispersive solid phase extraction of caffeine from exhaled breath condensate samples of premature infants prior to HPLC-PDA 异构体钴基金属有机框架和还原氧化石墨烯纳米复合材料用于在 HPLC-PDA 前从早产儿呼出气体冷凝物样品中分散固相萃取咖啡因

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124339

Samineh Raha , Ali Akbar Fathi , Mohammad Reza Afshar Mogaddam , Ali Shahedi-Hodjaghan , Mir Ali Farajzadeh , Mohamadbagher Hosseini , Maryam Khoubnasabjafari , Vahid Jouyban-Gharamaleki , Abolghasem Jouyban

{"title":"Heteroatom cobalt-based metal-organic framework and reduced graphene oxide nanocomposite for dispersive solid phase extraction of caffeine from exhaled breath condensate samples of premature infants prior to HPLC-PDA","authors":"Samineh Raha , Ali Akbar Fathi , Mohammad Reza Afshar Mogaddam , Ali Shahedi-Hodjaghan , Mir Ali Farajzadeh , Mohamadbagher Hosseini , Maryam Khoubnasabjafari , Vahid Jouyban-Gharamaleki , Abolghasem Jouyban","doi":"10.1016/j.jchromb.2024.124339","DOIUrl":"10.1016/j.jchromb.2024.124339","url":null,"abstract":"<div><div>A cobalt-based metal–organic framework and graphene oxide were combined to prepare a new nanocomposite for extracting of caffeine from exhaled breath condensate (EBC) samples. Dispersive micro solid phase extraction of caffeine was conducted using the nanocomposite as a sorbent by adding 10 mg of it to the sample solution and vortexing for 3 min. After extracting of the analyte, it was eluted using the mobile phase. The analyte was then analyzed using high performance liquid chromatography-photodiode array detector. Under optimal conditions, the limit of detection, limit of quantification, and linear range of the calibration curve were found to be 1.7, 5.9, and 10–500 µg/L, respectively. To assess the precision of the method, five replicates of standard solutions containing caffeine at two different concentration levels (50 and 100 µg/L) were tested. The relative standard deviations for intra- and inter-day precisions ranged from 4.3 to 6.8 %. The applicability of the method was demonstrated by analyzing the samples obtained from premature infants undergoing caffeine treatment and caffeine concentrations were 4.9 ± 0.6, 2.7 ± 0.2 µg/L in the EBC samples of who were under treatment by a 5-mg dose. Also, caffeine concentrations were 5.9 ± 0.3 and 18 ± 0.6 µg/L in the the infants who obtained the 10-mg and 25-mg doses, respectively. The results indicated a satisfactory, extraction recovery of 86 % showcasing the method’s reliability and effectiveness in analyzing real samples.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124339"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Pharmacokinetic profiling and network pharmacology of honey-fried Licorice: An Integrative workflow to study traditional Chinese medicines (TCMs) 蜜炒甘草的药代动力学分析和网络药理学：研究传统中药 (TCMs) 的综合工作流程。

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124353

Lifeng Zhao , Xin Yu , Siyang Wu , Kexin Xia , Yuyan Wang , Peichong Qin , Zhishan Huang , Chen Kang , Zheng Yuan , Yingfei Li

{"title":"Pharmacokinetic profiling and network pharmacology of honey-fried Licorice: An Integrative workflow to study traditional Chinese medicines (TCMs)","authors":"Lifeng Zhao , Xin Yu , Siyang Wu , Kexin Xia , Yuyan Wang , Peichong Qin , Zhishan Huang , Chen Kang , Zheng Yuan , Yingfei Li","doi":"10.1016/j.jchromb.2024.124353","DOIUrl":"10.1016/j.jchromb.2024.124353","url":null,"abstract":"<div><div>Licorice, known as the “elder statesman,” is commonly used in traditional Chinese medicine (TCM) formulations. This study aims to establish a workflow combining animal and in silico experiments to elucidate the mechanisms of TCMs at both qualitative and quantitative levels. UPLC-Q-TOF-MS/MS was employed to qualitatively characterize the total components of honey-fried licorice and the plasma components after oral administration in Beagle dogs. A UPLC-Q-Trap-MS/MS method was developed for the pharmacokinetic study of honey-fried licorice components in Beagle dog plasma. Network pharmacology and molecular docking were utilized to explore the primary functional targets and pathways. In total, we identified 68 constituents in honey-fried licorice, with 28 detected in Beagle dog plasma, and 18 of them, mainly belong to flavonoids and terpenoids, showing significant exposure. The plasma pharmacokinetic study of these 18 constituents revealed that compounds like liquiritin, glycyrrhizic acid, licoricesaponin G2, and glycyrrhetic acid-3-o-glucuronide had significant exposure. Network pharmacology and molecular docking analyses identified MAPK3, PIK3CB, PIK3CA, RAF1, and EGFR as the main targets of the active constituents of honey-fried licorice, involved in pathways such as the Ras signaling pathway, human cytomegalovirus infection, and the MAPK signaling pathway. This study provides a comprehensive profile and pharmacokinetic characteristics of honey-fried licorice, offering insights into its pharmacological, toxicological, and clinical aspects. The established workflow can serve as a standard for investigating other TCMs.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124353"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142611269","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Deciphering the impact and mechanism of total flavonoids from Cortex Juglandis Mandshuricae on alcoholic fatty liver employing LC-MS/MS, network pharmacology analysis and in vitro validation 通过LC-MS/MS、网络药理学分析和体外验证，破译Cortex Juglandis Mandshuricae总黄酮对酒精性脂肪肝的影响和机制

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124334

Tianmei Niu , Jiaxin Wang , Liying Xun , Bingqing Zheng , Zhipeng Deng , Zhi Chen , Kaijie Jia , Pan Zhao , Qitao Zhao

{"title":"Deciphering the impact and mechanism of total flavonoids from Cortex Juglandis Mandshuricae on alcoholic fatty liver employing LC-MS/MS, network pharmacology analysis and in vitro validation","authors":"Tianmei Niu , Jiaxin Wang , Liying Xun , Bingqing Zheng , Zhipeng Deng , Zhi Chen , Kaijie Jia , Pan Zhao , Qitao Zhao","doi":"10.1016/j.jchromb.2024.124334","DOIUrl":"10.1016/j.jchromb.2024.124334","url":null,"abstract":"<div><div>The Cortex Juglandis Mandshuricae (CJM) has the efficacy of penetrating the liver meridian, removing heat and dampness, and alleviating the liver, which corresponds to the pathogenesis of alcoholic fatty liver disease (AFLD) with damp heat accumulation. Modern research has shown that total flavonoids from Cortex Juglandis Mandshuricae (TFC) have hepatoprotective, antioxidant and antitumour pharmacological effects. However, there is no any investigation on the mechanism of TFC improving AFLD. In this work, a valid strategy combining UPLC-Q-Exactive Orbitrap-MS, network pharmacology and in vitro cellular experimental validation is proposed to predict the targets and pathways of TFC to ameliorate AFLD and to explore its mechanism of action. As a result, 26 flavonoids and 182 targets linked to TFC and AFLD were identified. These compounds realize their critical targets via various signaling pathways and perform multiple biological functions on the basis of the constructed compound-disease target networks. In vitro experiments demonstrated TFC had a protective impact on ethanol-treated L02 cells to a certain extent and could diminished lipid accretion. In addition, RT-qPCR and western blot results illustrated that TFC could regulate the expression of PPARα, CPT-1, SREBP-1c and FAS, and inhibit alcohol-induced lipid accumulation in L02 cells thereby alleviating AFLD. The present study further provides experimental justification for TFC to ameliorate AFLD in practical applications.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124334"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantitative analysis of methylmalonic acid in human plasma by LC-MS/MS after derivatization 衍生化后通过 LC-MS/MS 对人体血浆中的甲基丙二酸进行定量分析

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124359

Danhong Shen , Wei Chen , Jindi Liu , Yanhua Liu , Hongjun Shi

{"title":"Quantitative analysis of methylmalonic acid in human plasma by LC-MS/MS after derivatization","authors":"Danhong Shen , Wei Chen , Jindi Liu , Yanhua Liu , Hongjun Shi","doi":"10.1016/j.jchromb.2024.124359","DOIUrl":"10.1016/j.jchromb.2024.124359","url":null,"abstract":"<div><div>Methylmalonic acid (MMA) is a reverse biomarker of vitamin B12 that is increasingly utilized in clinical practice. However, its low sensitivity and susceptibility to strong interference from isomer present chromatographic challenges. We have developed a rapid derivatization method for plasma MMA at room temperature, converting it to the corresponding 2,2,2-trifluoroethylamide derivative using N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) and 2,2,2-trifluoroethylamine hydrochloride (TFEA). Amidization was completed within 10 min, followed by protein precipitation extraction of the amides with trichloroacetic acid for Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. This technique notably enhanced the signal-to-noise ratio of MMA in chromatography. The derivatized MMA exhibited excellent linearity within a concentration range of 42.4–2711.9 nmol/L, with a correlation coefficient (R<sup>2</sup>) of 0.9990. The intraday and interday precision of replicate measurements ranged from 2.4 % to 4.4 % and 2.6 % to 2.8 %, respectively, while the recovery fell between 97.9 % and 100.1 %.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124359"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142586148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of the ionization efficiency in phosphatidylcholine positional isomers with docosahexaenoic acid bound to the sn-1 or sn-2 position 评估与二十二碳六烯酸结合在 sn-1 或 sn-2 位置的磷脂酰胆碱位置异构体的电离效率。

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124355

Kana Fujiwara , Seiya Tanaka , Koyama Tomoyuki , Kazuaki Yoshinaga , Naohiro Gotoh

{"title":"Evaluation of the ionization efficiency in phosphatidylcholine positional isomers with docosahexaenoic acid bound to the sn-1 or sn-2 position","authors":"Kana Fujiwara , Seiya Tanaka , Koyama Tomoyuki , Kazuaki Yoshinaga , Naohiro Gotoh","doi":"10.1016/j.jchromb.2024.124355","DOIUrl":"10.1016/j.jchromb.2024.124355","url":null,"abstract":"<div><div>Phosphatidylcholine (PC), a key phospholipid, contains 2 fatty acids that can be bound at the sn-1 and sn-2 positions, resulting in positional isomers when different fatty acids are attached. Currently, there is no established method for identifying phospholipid molecular species and quantifying individual isomers using authentic standards of each PC isomer. In this study, we prepare authentic analytical standards for PC positional isomers through chemical synthesis and preparative purification. These isomers contain docosahexaenoic acid (DHA, 22:6) and palmitic acid (16:0) attached at the sn-1 and sn-2 positions and are denoted as PC(22:6/16:0) and PC(16:0/22:6), respectively. Standard solutions of PC(22:6/16:0) and PC(16:0/22:6) were analyzed using liquid chromatography-tandem mass spectrometry, and calibration curves of the PC positional isomers were generated to compare their ionization efficiencies. The ionization efficiency of PC(22:6/16:0) was 2.32 times higher than that of PC(16:0/22:6), indicating that the ionization efficiency depends on the binding position of the fatty acid. Elucidating and correcting the differences in the ionization efficiencies of the PC positional isomers will enable the accurate quantitative analysis of lipidomes in the future.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124355"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142643556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synthesis of 2-methyl-6-methoxy-4-quinolinecarboxylic acid N-hydroxysuccinimide ester (MMQC-OSu) for streamlined and effective HPLC-based fluorescence detection of aliphatic amines in environmental samples 合成 2-甲基-6-甲氧基-4-喹啉羧酸 N-羟基琥珀酰亚胺酯 (MMQC-OSu)，用于基于高效液相色谱法的荧光检测环境样品中的脂肪胺类化合物

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124348

Jian Fei , Qiuyue Sha , Wenjie Zhu , Si Liu , Zhaoyu Hu , Wei Du , Xin Liu

{"title":"Synthesis of 2-methyl-6-methoxy-4-quinolinecarboxylic acid N-hydroxysuccinimide ester (MMQC-OSu) for streamlined and effective HPLC-based fluorescence detection of aliphatic amines in environmental samples","authors":"Jian Fei , Qiuyue Sha , Wenjie Zhu , Si Liu , Zhaoyu Hu , Wei Du , Xin Liu","doi":"10.1016/j.jchromb.2024.124348","DOIUrl":"10.1016/j.jchromb.2024.124348","url":null,"abstract":"<div><div>Aliphatic amines are widely distributed in the environment and food sources, posing potential health risks through skin and mucosal irritation. Consequently, their quantitative detection is crucial for assessing environmental health. Despite high reactivity and fluorescence properties of succinimidyl ester-based derivatization reagents, their application in the aliphatic amines detection is hampered by challenges such as limited detection sensitivity and fluorescence interference. We established an innovative synthetic approach to produce a series of succinimidyl esters with the desirable substituents. This advancement enabled the creation of efficient and highly sensitive reagents for the detection of aliphatic amines. Among them, 2-methyl-6-methoxy-4-quinolinecarboxylic acid N-hydroxysuccinimide ester (MMQC-OSu) exhibited the best detection performance. MMQC-OSu reacted with aliphatic amines at 40 °C in pH 8.0 buffer for 20 min, which was subsequently separated in a C18 chromatographic column with the fluorescence detection wavelength of 336/432 nm. This detection approach featured a rapid and mild reaction process, minimal interference from corresponding hydrolysis products, and impressive sensitivity (0.05 nM). These characteristics indicate that MMQC-OSu significantly surpassing commercial aliphatic amines detection reagents. Finally, the detection strategy of MMQC-OSu was successfully applied in environmental sample analysis, with recovery rate of 93 %–108 % and RSDs between 1.4 % and 6.5 %.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124348"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142592594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of imaged capillary isoelectric focusing as a platform mispairing byproduct testing method for asymmetric WuXiBody-based bispecific antibodies 开发成像毛细管等电聚焦技术，作为基于非对称武溪体的双特异性抗体的平台误配对副产物检测方法

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124357

Liping Zhan , Luping Xie , Fujiao Lv , Jincui Huang , Zhi Jian Chen , Li Wang , Zhiqiang Chen

{"title":"Development of imaged capillary isoelectric focusing as a platform mispairing byproduct testing method for asymmetric WuXiBody-based bispecific antibodies","authors":"Liping Zhan , Luping Xie , Fujiao Lv , Jincui Huang , Zhi Jian Chen , Li Wang , Zhiqiang Chen","doi":"10.1016/j.jchromb.2024.124357","DOIUrl":"10.1016/j.jchromb.2024.124357","url":null,"abstract":"<div><div>This study explores the application of imaged capillary isoelectric focusing (icIEF) to distinguish and quantify mispairing byproducts in asymmetric WuXiBody-based bispecific antibodies (AsWXbsAbs). Bispecific antibody (BsAb), developed using Knobs-into-Holes (KiH) technology, often result in byproducts such as knob-knob (KK) and hole-hole (HH) homodimers, which share similar sizes with the target BsAb, complicating their separation by traditional methods like size exclusion chromatography-high performance liquid chromatography (SEC-HPLC). Our approach leverages the unique pI differences introduced by substituting the CH1/CL domain with the T cell receptor (TCR) constant domain in AsWXbsAbs. This modification enables icIEF to effectively differentiate between the KK and HH homodimers and the target BsAb. Through the construction and expression of heavy and light chain variants, we validated that the experimental pI values aligned with theoretical predictions, confirming icIEF’s capability in distinguishing these entities. Enrichment of in-process K-related and H-related species was achieved, allowing for high-purity samples necessary for icIEF method development. The method was qualified and showed good specificity and linearity, with a quantitation limit of 4 % for K-related species (R<sup>2</sup> = 0.9919) and 1 % for H-related species (R<sup>2</sup> = 0.9805). This method was used effectively as an in-process test and release assay, proving its simple and quick utility in multiple AsWXbsAbs projects.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124357"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142586149","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantifying the extended energy metabolome of industrially important microorganisms (Saccharomyces cerevisiae) using ultra-performance liquid chromatography with mass spectrometry 利用超高效液相色谱-质谱法量化具有工业重要性的微生物（酿酒酵母）的扩展能量代谢组

IF 2.8 3区医学

Journal of Chromatography B Pub Date : 2024-11-01 DOI: 10.1016/j.jchromb.2024.124342

Jordan I. Oliver, Antony N. Davies, Richard Dinsdale

{"title":"Quantifying the extended energy metabolome of industrially important microorganisms (Saccharomyces cerevisiae) using ultra-performance liquid chromatography with mass spectrometry","authors":"Jordan I. Oliver, Antony N. Davies, Richard Dinsdale","doi":"10.1016/j.jchromb.2024.124342","DOIUrl":"10.1016/j.jchromb.2024.124342","url":null,"abstract":"<div><div>This study has developed a new targeted methodology for the separation, detection, and quantification of metabolites from the wider energy metabolome of industrially important microorganisms such as <em>Saccharomyces cerevisiae</em> in a single analytical sample. This has been achieved using UHPLC-MS technology in HILIC mode. Absolute concentrations of metabolites nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide reduced (NADH), nicotinamide adenine dinucleotide phosphate (NADP), nicotinamide adenine dinucleotide phosphate reduced (NADPH), flavin adenine dinucleotide (FAD), adenosine-monophosphate (AMP), adenosine-diphosphate (ADP), and adenosine-triphosphate (ATP) were determined in a single extraction and analytical methodology.</div><div>This study demonstrated the development of a rapid, statistically robust, and reproducible methodology through regression calibrations of standard samples from 0.1 to 100 µMol providing a correlation value of r<sup>2</sup> = >0.98 for all metabolites. Sample preparation, extraction and analytical methodologies used showed high accuracy, sensitivity, and recovery. With an LOD and LOQ for the targeted analysis of metabolites from the wider energy metabolism in a single sample and analytical run with the lowest LOD of 0.055 nMol (±0.002) and lowest LOQ of 0.167 nMol (±0.006). This method was then applied to <em>Saccharomyces cerevisiae</em> cell culture to evaluate the methodology in industrially used microbial cultures. Results obtained have been statistically determined to be robust and reproducible through recovery analysis using deuterated and isotopically labelled internal standards AMP-<sup>15</sup>N, ADP-<sup>15</sup>N and ATP-d<sub>14</sub>.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1248 ","pages":"Article 124342"},"PeriodicalIF":2.8,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142560707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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