Pub Date : 2025-11-01Epub Date: 2025-07-31DOI: 10.1016/j.jchromb.2025.124747
S Chevolleau, C Orlandi, L Mervant, R Vuillaume, I Jouanin, N Naud, F Pierre, F Gueraud, L Debrauwer
The elevated risk of colorectal cancer (CRC) induced by red or processed meat rich diets is now established. Those haem‑iron rich diets induce luminal lipid peroxidation, one of the most recognised hypotheses explaining CRC promotion. Due to their known toxic properties, quantification of reactive aldehydes such as 4-hydroxy-2(E)-nonenal (HNE) and 4-hydroxy-2(E)-hexenal (HHE) as lipid peroxidation end-products in biological fluids is of upmost importance. Following previous works on faecal waters, an UHPLC-ESI-MS/MS method has been developed and validated for HNE and HHE quantification in rat serum, using deuterated internal standards (ISs). After protein precipitation (PP) and solid phase extraction (SPE), LC-ESI-MS/MS analysis was achieved by MRM. The use of a brominated derivatisation reagent allowed using the bromine isotopes for selective detection of both HNE and HHE based on diagnostic transitions. This new method was validated according to the European Medicines Agency (EMA) guidelines. Our method proved to efficiently determine HNE and HHE serum concentrations with the required sensitivity (nM range) in serum of rats fed diets rich or not in red meat and different fatty acid compositions.
{"title":"Significant improvements in targeted UHPLC-ESI-MS/MS analysis of the reactive aldehydes 4-hydroxy-2(E)-nonenal and 4-hydroxy-2(E)-hexenal and application to rat serum.","authors":"S Chevolleau, C Orlandi, L Mervant, R Vuillaume, I Jouanin, N Naud, F Pierre, F Gueraud, L Debrauwer","doi":"10.1016/j.jchromb.2025.124747","DOIUrl":"10.1016/j.jchromb.2025.124747","url":null,"abstract":"<p><p>The elevated risk of colorectal cancer (CRC) induced by red or processed meat rich diets is now established. Those haem‑iron rich diets induce luminal lipid peroxidation, one of the most recognised hypotheses explaining CRC promotion. Due to their known toxic properties, quantification of reactive aldehydes such as 4-hydroxy-2(E)-nonenal (HNE) and 4-hydroxy-2(E)-hexenal (HHE) as lipid peroxidation end-products in biological fluids is of upmost importance. Following previous works on faecal waters, an UHPLC-ESI-MS/MS method has been developed and validated for HNE and HHE quantification in rat serum, using deuterated internal standards (ISs). After protein precipitation (PP) and solid phase extraction (SPE), LC-ESI-MS/MS analysis was achieved by MRM. The use of a brominated derivatisation reagent allowed using the bromine isotopes for selective detection of both HNE and HHE based on diagnostic transitions. This new method was validated according to the European Medicines Agency (EMA) guidelines. Our method proved to efficiently determine HNE and HHE serum concentrations with the required sensitivity (nM range) in serum of rats fed diets rich or not in red meat and different fatty acid compositions.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1265 ","pages":"124747"},"PeriodicalIF":2.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811470","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}
Objectives: The Yangluan Formula (YLF) influences the outcomes of in vitro fertilization-embryo transfer (IVF-ET) in infertile women with diminished ovarian reserve (DOR); however, the potential mechanisms by which YLF ameliorates DOR have not yet been elucidated. The aim of this study was to examine the effects of YLF on IVF-ET outcomes in infertile women with DOR, and to elucidate the potential mechanisms by which YLF addresses DOR.
Methods: Non-targeted metabolomics studies were conducted on follicular fluid specimens procured from individuals with DOR treated with or without YLF, and from patients with normal ovarian reserve who underwent IVF-ET treatment. Distinct metabolites were identified using untargeted metabolomics, and MetaboAnalyst was used to examine metabolic pathways. After applying network pharmacology (NP), the target of YLF acting on DOR was determined. Cytoscape software was used to develop compound-reaction-enzyme-gene networks, and molecular docking (MD) simulations were conducted to confirm the link between YLF and crucial targets.
Results: Patients with DOR showed a notable reduction in the number of oocytes retrieved, incidence of 2PN fertilization, and number of cleaved embryos (P < 0.001). Additionally, the DOR cohort exhibited a markedly reduced quantity of high-quality embryos on day 3 compared to the CON cohort (P < 0.005). In contrast to the DOR cohort, the YLF cohort exhibited notably superior outcomes in terms of 2PN fertilization rates, cleavage-stage embryo development, and the number of high-grade embryos on day 3 (P < 0.05). The proportion of 2PN fertilization observed in YLF subjects substantially exceeded that in DOR individuals (81.2 % vs. 64.3 %, P < 0.05). Combined analysis of metabolomics and NP, focusing on five key targets for the action of YLF (monoamine oxidase A, monoamine oxidase B, myeloperoxidase, xanthine dehydrogenase, and phosphodiesterase 3A), four key metabolites (pelargonic acid, 1-(5-Phospho-D-ribosyl)-5-amino-4-imidazolecarboxylate, isokobusone, 5-O-(1-Carboxyvinyl)-3-phosphoshikimate), and two related pathways (glycine, serine, alanine, and threonine metabolism).
Conclusion: We elucidated the mode of action of YLF in DOR treatment by integrating metabolomics and NP. YLF can effectively improve IVF outcomes in patients with DOR. This study provides new perspectives on the mechanism by which YLF improves ovarian function.
{"title":"Integrating network pharmacology and metabolomics to elucidate the mechanism of action of Yangluan formula for treating of diminished ovarian reserve.","authors":"Yang Wang, Panwei Hu, Hua Yan, Yuanyuan Wu, Ping Yin, Dongyi Shen, Xiaole Zhang, Cong Qi, Qinhua Zhang","doi":"10.1016/j.jchromb.2025.124749","DOIUrl":"10.1016/j.jchromb.2025.124749","url":null,"abstract":"<p><strong>Objectives: </strong>The Yangluan Formula (YLF) influences the outcomes of in vitro fertilization-embryo transfer (IVF-ET) in infertile women with diminished ovarian reserve (DOR); however, the potential mechanisms by which YLF ameliorates DOR have not yet been elucidated. The aim of this study was to examine the effects of YLF on IVF-ET outcomes in infertile women with DOR, and to elucidate the potential mechanisms by which YLF addresses DOR.</p><p><strong>Methods: </strong>Non-targeted metabolomics studies were conducted on follicular fluid specimens procured from individuals with DOR treated with or without YLF, and from patients with normal ovarian reserve who underwent IVF-ET treatment. Distinct metabolites were identified using untargeted metabolomics, and MetaboAnalyst was used to examine metabolic pathways. After applying network pharmacology (NP), the target of YLF acting on DOR was determined. Cytoscape software was used to develop compound-reaction-enzyme-gene networks, and molecular docking (MD) simulations were conducted to confirm the link between YLF and crucial targets.</p><p><strong>Results: </strong>Patients with DOR showed a notable reduction in the number of oocytes retrieved, incidence of 2PN fertilization, and number of cleaved embryos (P < 0.001). Additionally, the DOR cohort exhibited a markedly reduced quantity of high-quality embryos on day 3 compared to the CON cohort (P < 0.005). In contrast to the DOR cohort, the YLF cohort exhibited notably superior outcomes in terms of 2PN fertilization rates, cleavage-stage embryo development, and the number of high-grade embryos on day 3 (P < 0.05). The proportion of 2PN fertilization observed in YLF subjects substantially exceeded that in DOR individuals (81.2 % vs. 64.3 %, P < 0.05). Combined analysis of metabolomics and NP, focusing on five key targets for the action of YLF (monoamine oxidase A, monoamine oxidase B, myeloperoxidase, xanthine dehydrogenase, and phosphodiesterase 3A), four key metabolites (pelargonic acid, 1-(5-Phospho-D-ribosyl)-5-amino-4-imidazolecarboxylate, isokobusone, 5-O-(1-Carboxyvinyl)-3-phosphoshikimate), and two related pathways (glycine, serine, alanine, and threonine metabolism).</p><p><strong>Conclusion: </strong>We elucidated the mode of action of YLF in DOR treatment by integrating metabolomics and NP. YLF can effectively improve IVF outcomes in patients with DOR. This study provides new perspectives on the mechanism by which YLF improves ovarian function.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1265 ","pages":"124749"},"PeriodicalIF":2.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144797780","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}
Ensuring the authenticity and quality of Sida rhombifolia raw materials is crucial for its herbal medicinal product's consistent efficacy, quality, and safety. In this study, we developed an identification and authentication method for identifying and authenticating S. rhombifolia from Turnera subulata. T. subulata has the same leaf morphology as S. rhombifolia, so that it could be used as an adulteration raw material for S. rhombifolia. We employed liquid chromatography-high resolution mass spectrometry (LC-HRMS)- and thin-layer chromatography (TLC)-based metabolomics for that purpose. Distinct chemical fingerprints of S. rhombifolia from T. subulata were obtained using LC-HRMS and TLC fingerprint analysis. Mixtures of S. rhombifolia and T. subulata powdered samples at varying concentrations (5 %, 25 %, and 50 % w/w) were analyzed using principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to see differences of each group. The PCA score plot from the TLC analysis explained over 70 % of the total variance, while LC-HRMS data provided the highest classification accuracy at 97.05 %. This integrated approach enhances the reliability of S. rhombifolia authentication by combining TLC's rapid profiling capability with LC-HRMS's analytical precision. This study provides a robust analytical framework for the quality control of herbal medicines, specifically addressing challenges related to adulteration.
{"title":"LC-HRMS- and TLC-based metabolomics for the identification and authentication of Sida rhombifolia","authors":"Uswatun Hasanah , Eti Rohaeti , Irmanida Batubara , Utami Dyah Syafitri , Rudi Heryanto , Taopik Ridwan , Nancy Dewi Yuliana , Mohamad Rafi","doi":"10.1016/j.jchromb.2025.124834","DOIUrl":"10.1016/j.jchromb.2025.124834","url":null,"abstract":"<div><div>Ensuring the authenticity and quality of <em>Sida rhombifolia</em> raw materials is crucial for its herbal medicinal product's consistent efficacy, quality, and safety. In this study, we developed an identification and authentication method for identifying and authenticating <em>S. rhombifolia</em> from <em>Turnera subulata</em>. <em>T. subulata</em> has the same leaf morphology as <em>S. rhombifolia,</em> so that it could be used as an adulteration raw material for <em>S. rhombifolia</em>. We employed liquid chromatography-high resolution mass spectrometry (LC-HRMS)- and thin-layer chromatography (TLC)-based metabolomics for that purpose. Distinct chemical fingerprints of <em>S. rhombifolia</em> from <em>T. subulata</em> were obtained using LC-HRMS and TLC fingerprint analysis. Mixtures of <em>S. rhombifolia</em> and <em>T. subulata</em> powdered samples at varying concentrations (5 %, 25 %, and 50 % <em>w</em>/w) were analyzed using principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to see differences of each group. The PCA score plot from the TLC analysis explained over 70 % of the total variance, while LC-HRMS data provided the highest classification accuracy at 97.05 %. This integrated approach enhances the reliability of <em>S. rhombifolia</em> authentication by combining TLC's rapid profiling capability with LC-HRMS's analytical precision. This study provides a robust analytical framework for the quality control of herbal medicines, specifically addressing challenges related to adulteration.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124834"},"PeriodicalIF":2.8,"publicationDate":"2025-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145446933","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}
Pub Date : 2025-10-29DOI: 10.1016/j.jchromb.2025.124840
Lingxiao Zhang , Jie Qu , Xinyan Sun , Haiyan Dong , Hongping Yao , Xiaoliang Cheng
Pyrotinib which is a novel irreversible tyrosine kinase inhibitor plus docetaxel or paclitaxel is effective for patients with Her2 positive early or advanced breast cancer including those who failed in first-line treatment. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and verified for simultaneous quantification of pyrotinib, docetaxel and paclitaxel in human plasma, and applied to therapeutic drug monitoring. A reversed-phase Hypersil GOLD aQ column eluted by a gradient mobile phase composed of water and acetonitrile both containing 0.1 % formic acid under flow rate of 0.3 mL min−1 was used for chromatographic separation. The mass spectrometry was operated in positive electrospray ionization mode, and selective reaction monitoring was applied for quantitative analysis. With imatinib as internal standard, one-step deproteinization approach with acetonitrile was applied to extract analytes and purify specimens. This method was adequately validated according to guidelines in terms of specificity and selectivity, sensitivity, linearity, extraction recovery, matrix effect, precision and accuracy, dilution integration and stability. The validated method was applied to therapeutic drug monitoring for breast cancer patients receiving pyrotinib and taxanes based chemotherapy. The therapeutic drug monitoring results showed that the plasma concentration of pyrotinib, docetaxel and paclitaxel varied significantly among individuals. Therapeutic drug monitoring for pyrotinib, docetaxel and paclitaxel is essential for individualized treatment to ensure efficacy and safety.
Pyrotinib是一种新型的不可逆酪氨酸激酶抑制剂，与多西紫杉醇或紫杉醇联合使用对Her2阳性的早期或晚期乳腺癌患者有效，包括那些在一线治疗失败的患者。建立并验证了同时定量测定人血浆中吡罗替尼、多西他赛和紫杉醇的液相色谱-串联质谱（LC-MS/MS）方法，并将其应用于治疗药物监测。采用反相Hypersil GOLD aQ色谱柱，以含有0.1%甲酸的水和乙腈组成的梯度流动相洗脱，流速为0.3 mL min-1。质谱分析采用正电喷雾电离模式，定量分析采用选择性反应监测。以伊马替尼为内标，采用乙腈一步脱蛋白法提取分析物，纯化样品。本方法在特异性和选择性、灵敏度、线性度、提取回收率、基质效应、精密度和准确度、稀释积分和稳定性等方面均按照指南进行了充分验证。将验证的方法应用于以吡罗替尼和紫杉烷为基础的化疗的乳腺癌患者的治疗药物监测。治疗药物监测结果显示，吡罗替尼、多西他赛和紫杉醇的血药浓度在个体间差异显著。吡罗替尼、多西紫杉醇和紫杉醇的治疗药物监测对于个体化治疗至关重要，以确保疗效和安全性。
{"title":"An LC-MS/MS method for simultaneous quantification of pyrotinib, docetaxel and paclitaxel in human plasma and its application to therapeutic drug monitoring","authors":"Lingxiao Zhang , Jie Qu , Xinyan Sun , Haiyan Dong , Hongping Yao , Xiaoliang Cheng","doi":"10.1016/j.jchromb.2025.124840","DOIUrl":"10.1016/j.jchromb.2025.124840","url":null,"abstract":"<div><div>Pyrotinib which is a novel irreversible tyrosine kinase inhibitor plus docetaxel or paclitaxel is effective for patients with Her2 positive early or advanced breast cancer including those who failed in first-line treatment. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed and verified for simultaneous quantification of pyrotinib, docetaxel and paclitaxel in human plasma, and applied to therapeutic drug monitoring. A reversed-phase Hypersil GOLD aQ column eluted by a gradient mobile phase composed of water and acetonitrile both containing 0.1 % formic acid under flow rate of 0.3 mL min<sup>−1</sup> was used for chromatographic separation. The mass spectrometry was operated in positive electrospray ionization mode, and selective reaction monitoring was applied for quantitative analysis. With imatinib as internal standard, one-step deproteinization approach with acetonitrile was applied to extract analytes and purify specimens. This method was adequately validated according to guidelines in terms of specificity and selectivity, sensitivity, linearity, extraction recovery, matrix effect, precision and accuracy, dilution integration and stability. The validated method was applied to therapeutic drug monitoring for breast cancer patients receiving pyrotinib and taxanes based chemotherapy. The therapeutic drug monitoring results showed that the plasma concentration of pyrotinib, docetaxel and paclitaxel varied significantly among individuals. Therapeutic drug monitoring for pyrotinib, docetaxel and paclitaxel is essential for individualized treatment to ensure efficacy and safety.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124840"},"PeriodicalIF":2.8,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145440431","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}
Pub Date : 2025-10-28DOI: 10.1016/j.jchromb.2025.124830
Xi Hu , Siye Gao , Hancong Wu , Qingyun He , Jianing Liu , Danyue Zhao , Zhenqiang Wu
This study developed an environmentally friendly method for extracting polyphenols and rubusoside from Rubus suavissimus leaves. Under optimal extraction conditions, the contents of polyphenol and rubusoside reached 91.89 ± 2.39 mg/g and 47.03 ± 1.45 mg/g, respectively. The extract was purified by twelve types of macroporous resins, among which the ADS-7 resin had the highest static adsorption and desorption capacities. Optimized purification parameters for ADS-7 resin were determined by static and dynamic adsorption-desorption experiments and found to be: loading flow rate, 3 BV/h; loading concentration, 10 mg/mL; loading volume, 24 BV; elution solvent, 80 % (v/v) ethanol; elution speed, 3 BV/h; and elution volume, 20 BV. After purification, the content of polyphenols and rubusoside increased by 2.63 and 2.32 times, respectively. Kinetic and thermodynamic analyses revealed distinct adsorption mechanisms: polyphenols followed a pseudo-second-order model, whereas rubusoside followed a pseudo-first-order model; both exhibited spontaneous (ΔG < 0) and endothermic (ΔH > 0) physisorption.
Ultrafiltration-assisted centrifugal fractionation combined with UPLC-ESI-qTOF-MS/MS identified eight potential α-glucosidase inhibitors, including quercetin, apigenin, rutin, ellagic acid, kaempferol-3-O-rutinoside, kaempferol, rubusoside, and steviol. Molecular docking simulations revealed the molecular interactions between these compounds and α-glucosidase.
The enriched extract demonstrated enhanced bioactivity, with a 3.35-fold higher α-glucosidase inhibition, and its IC₅₀ values for DPPH and ABTS+ radical scavenging decreased from 142.03 μg/mL to 88.45 μg/mL and from 197.63 μg/mL to 107.63 μg/mL, respectively. These results substantiate the effectiveness of the extraction-purification approach for producing R. suavissimus leaf extracts and suggest their potential application as functional hypoglycemic and antioxidant food ingredients.
{"title":"Characterization and purification of hypoglycemic components from Rubus suavissimus leaf by affinity ultrafiltration, molecular docking, and macroporous resin","authors":"Xi Hu , Siye Gao , Hancong Wu , Qingyun He , Jianing Liu , Danyue Zhao , Zhenqiang Wu","doi":"10.1016/j.jchromb.2025.124830","DOIUrl":"10.1016/j.jchromb.2025.124830","url":null,"abstract":"<div><div>This study developed an environmentally friendly method for extracting polyphenols and rubusoside from <em>Rubus suavissimus</em> leaves. Under optimal extraction conditions, the contents of polyphenol and rubusoside reached 91.89 ± 2.39 mg/g and 47.03 ± 1.45 mg/g, respectively. The extract was purified by twelve types of macroporous resins, among which the ADS-7 resin had the highest static adsorption and desorption capacities. Optimized purification parameters for ADS-7 resin were determined by static and dynamic adsorption-desorption experiments and found to be: loading flow rate, 3 BV/h; loading concentration, 10 mg/mL; loading volume, 24 BV; elution solvent, 80 % (<em>v</em>/v) ethanol; elution speed, 3 BV/h; and elution volume, 20 BV. After purification, the content of polyphenols and rubusoside increased by 2.63 and 2.32 times, respectively. Kinetic and thermodynamic analyses revealed distinct adsorption mechanisms: polyphenols followed a pseudo-second-order model, whereas rubusoside followed a pseudo-first-order model; both exhibited spontaneous (ΔG < 0) and endothermic (ΔH > 0) physisorption.</div><div>Ultrafiltration-assisted centrifugal fractionation combined with UPLC-ESI-qTOF-MS/MS identified eight potential α-glucosidase inhibitors, including quercetin, apigenin, rutin, ellagic acid, kaempferol-3-O-rutinoside, kaempferol, rubusoside, and steviol. Molecular docking simulations revealed the molecular interactions between these compounds and α-glucosidase.</div><div>The enriched extract demonstrated enhanced bioactivity, with a 3.35-fold higher α-glucosidase inhibition, and its IC₅₀ values for DPPH and ABTS<sup>+</sup> radical scavenging decreased from 142.03 μg/mL to 88.45 μg/mL and from 197.63 μg/mL to 107.63 μg/mL, respectively. These results substantiate the effectiveness of the extraction-purification approach for producing <em>R. suavissimus</em> leaf extracts and suggest their potential application as functional hypoglycemic and antioxidant food ingredients.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124830"},"PeriodicalIF":2.8,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415411","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}
Pub Date : 2025-10-28DOI: 10.1016/j.jchromb.2025.124839
Yan Zhang, Zexi Yu, Xiaotong Zhu, Rongyue Zhang, Nan Li, Min Wang, Juan Qiao
A strategy utilizing oriented γ-cyclodextrin-based metal organic frameworks (γ-CD-MOFs) as chiral additives in micellar electrokinetic chromatography (MEKC) is presented for enhanced enantioseparation of dansylated D,L-amino acids (Dns-D,L-AAs). Its core innovation resides in the fixed orientation of γ-CD within the MOFs structure. This unique characteristic facilitates a shift in chiral recognition from “random interactions” to a “directional and highly efficient recognition” process, thereby substantially enhancing the resolution of chiral separation while overcoming the limitations of free γ-CD, including unstable interactions and poor reproducibility. Systematic optimization of SDS concentration (9.0 mM), γ-CD-MOFs concentration (16.0 mM), and buffer pH (9.5) yielded a maximum resolution (Rs) of 2.2, representing a 50 % improvement over traditional γ-CD. Under these conditions, 14 pairs of Dns-D,L-AAs achieved baseline separation, with 4 additional pairs showing partial separation. Quantitative validation for D,L-methionine (D,L-Met) demonstrated excellent linearity (19.8–1500 μM, r2 = 0.999), low limits of detection (6.6 μM) and quantitation (19.8 μM), and high stability (relative standard deviations <5 %). Application to rat plasma samples revealed peak concentration (Cmax) of D,L-Met at 90 min post-injection, highlighting its utility in pharmacokinetic studies of amino acid-derived prodrugs (e.g., S-adenosylmethionine). This work introduces oriented γ-CD-MOFs as a transformative chiral additive in MEKC, offering superior selectivity and reproducibility for pharmaceutical and biomedical analyses.
提出了一种利用定向γ-环糊精基金属有机骨架（γ-CD-MOFs）作为胶束电动色谱（MEKC）手性添加剂的策略，以增强丹化D， l -氨基酸（dn -D,L-AAs）的对端分离。其核心创新在于γ-CD在mof结构中的固定取向。这种独特的特性有助于手性识别从“随机相互作用”向“定向和高效识别”过程的转变，从而大大提高了手性分离的分辨率，同时克服了游离γ-CD的局限性，包括相互作用不稳定和再现性差。系统优化SDS浓度（9.0 mM）、γ-CD- mof浓度（16.0 mM）和缓冲液pH(9.5)，最大分辨率（Rs）为2.2，比传统的γ-CD提高了50%。在这些条件下，14对Dns-D， l - aa达到基线分离，另外4对部分分离。定量验证结果表明，D， l -蛋氨酸（D,L-Met）具有良好的线性关系（19.8 ~ 1500 μM, r2 = 0.999），低检出限（6.6 μM）和定量限（19.8 μM），高稳定性（相对标准偏差<； 5%）。应用于大鼠血浆样品显示，注射后90分钟D，L-Met的峰值浓度（Cmax），突出了其在氨基酸衍生的前药（如s -腺苷蛋氨酸）的药代动力学研究中的应用。本研究介绍了定向γ- cd - mof作为MEKC中的一种变革性手性添加剂，为药物和生物医学分析提供了优越的选择性和可重复性。
{"title":"γ-cyclodextrin based metal organic frameworks as micellar electrokinetic chromatography additive for the chiral separation of dansylated amino acids","authors":"Yan Zhang, Zexi Yu, Xiaotong Zhu, Rongyue Zhang, Nan Li, Min Wang, Juan Qiao","doi":"10.1016/j.jchromb.2025.124839","DOIUrl":"10.1016/j.jchromb.2025.124839","url":null,"abstract":"<div><div>A strategy utilizing oriented γ-cyclodextrin-based metal organic frameworks (γ-CD-MOFs) as chiral additives in micellar electrokinetic chromatography (MEKC) is presented for enhanced enantioseparation of dansylated D,L-amino acids (Dns-D,L-AAs). Its core innovation resides in the fixed orientation of γ-CD within the MOFs structure. This unique characteristic facilitates a shift in chiral recognition from “random interactions” to a “directional and highly efficient recognition” process, thereby substantially enhancing the resolution of chiral separation while overcoming the limitations of free γ-CD, including unstable interactions and poor reproducibility. Systematic optimization of SDS concentration (9.0 mM), γ-CD-MOFs concentration (16.0 mM), and buffer pH (9.5) yielded a maximum resolution (<em>R</em><sub>s</sub>) of 2.2, representing a 50 % improvement over traditional γ-CD. Under these conditions, 14 pairs of Dns-D,L-AAs achieved baseline separation, with 4 additional pairs showing partial separation. Quantitative validation for D,L-methionine (D,L-Met) demonstrated excellent linearity (19.8–1500 μM, r<sup>2</sup> = 0.999), low limits of detection (6.6 μM) and quantitation (19.8 μM), and high stability (relative standard deviations <5 %). Application to rat plasma samples revealed peak concentration (C<sub>max</sub>) of D,L-Met at 90 min post-injection, highlighting its utility in pharmacokinetic studies of amino acid-derived prodrugs (e.g., S-adenosylmethionine). This work introduces oriented γ-CD-MOFs as a transformative chiral additive in MEKC, offering superior selectivity and reproducibility for pharmaceutical and biomedical analyses.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124839"},"PeriodicalIF":2.8,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415410","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}
Pub Date : 2025-10-28DOI: 10.1016/j.jchromb.2025.124828
Qinyu Xiao , Jiamin Yang , Jianjun Xie , Qing Liu , Hongbo Huang , Yiwen Tao , Bo Ding
Metabolomics workflows involve multiple complex steps including sample collection, storage, preparation, metabolite extraction, analytical platforms selection, data acquisition and interpretation. Each step may introduce variability that affects the quality and reliability of metabolomic data. To systematically investigate the effects of these factors on metabolomics outcomes, plasma samples from four different anatomical sites of colon cancer patients were analyzed using Liquid chromatography- Quadrupole-Exactive Orbitrap mass spectrometry (LC-Q-Exactive Orbitrap MS) for untargeted metabolomics. Response surface methodology was employed to optimize the ultrasound-assisted extraction conditions during sample pretreatment. Data analysis strategies were systematically evaluated, including Feature-Based Molecular Networking (FBMN) construction parameters and comparative assessment of different FBMN platforms for metabolite annotation. The optimized extraction conditions were determined as 300 % methanol concentration, sample freezing at −20 °C for 40 min, followed by ultrasonication for 5 min. Sample standardization protocols requiring single-use portioning and limiting freeze-thaw cycles to ≤2–3 cycles were identified as essential for reliable biomarker discovery and therapeutic mechanism exploration. Optimal FBMN construction parameters comprised a 25-min gradient elution time, 50 mm chromatographic column length, and high sample concentration. Comparative evaluation of Global Natural Products Social Molecular Networking (GNPS) and MZmine implementations of FBMN revealed that GNPS was recommended for studies prioritizing comprehensive annotation coverage and discovery-oriented metabolomics, while MZmine was preferred for method development, or applications requiring local processing without external data upload. This study demonstrated that preprocessing and data analysis strategies were critical determinants of data quality in untargeted plasma metabolomics. The findings provided evidence-based recommendations for experimental design, storage conditions, and data handling procedures that can guide protocol standardization and minimize undesired analytical variation in metabolomics studies.
{"title":"Impact of Preprocessing and data analysis strategies on metabolite annotation in biological samples: A mass spectrometry-based metabolomics study using feature-based molecular networking","authors":"Qinyu Xiao , Jiamin Yang , Jianjun Xie , Qing Liu , Hongbo Huang , Yiwen Tao , Bo Ding","doi":"10.1016/j.jchromb.2025.124828","DOIUrl":"10.1016/j.jchromb.2025.124828","url":null,"abstract":"<div><div>Metabolomics workflows involve multiple complex steps including sample collection, storage, preparation, metabolite extraction, analytical platforms selection, data acquisition and interpretation. Each step may introduce variability that affects the quality and reliability of metabolomic data. To systematically investigate the effects of these factors on metabolomics outcomes, plasma samples from four different anatomical sites of colon cancer patients were analyzed using Liquid chromatography- Quadrupole-Exactive Orbitrap mass spectrometry (LC-Q-Exactive Orbitrap MS) for untargeted metabolomics. Response surface methodology was employed to optimize the ultrasound-assisted extraction conditions during sample pretreatment. Data analysis strategies were systematically evaluated, including Feature-Based Molecular Networking (FBMN) construction parameters and comparative assessment of different FBMN platforms for metabolite annotation. The optimized extraction conditions were determined as 300 % methanol concentration, sample freezing at −20 °C for 40 min, followed by ultrasonication for 5 min. Sample standardization protocols requiring single-use portioning and limiting freeze-thaw cycles to ≤2–3 cycles were identified as essential for reliable biomarker discovery and therapeutic mechanism exploration. Optimal FBMN construction parameters comprised a 25-min gradient elution time, 50 mm chromatographic column length, and high sample concentration. Comparative evaluation of Global Natural Products Social Molecular Networking (GNPS) and MZmine implementations of FBMN revealed that GNPS was recommended for studies prioritizing comprehensive annotation coverage and discovery-oriented metabolomics, while MZmine was preferred for method development, or applications requiring local processing without external data upload. This study demonstrated that preprocessing and data analysis strategies were critical determinants of data quality in untargeted plasma metabolomics. The findings provided evidence-based recommendations for experimental design, storage conditions, and data handling procedures that can guide protocol standardization and minimize undesired analytical variation in metabolomics studies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124828"},"PeriodicalIF":2.8,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415359","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}
Pub Date : 2025-10-26DOI: 10.1016/j.jchromb.2025.124837
Zhanying Chang , Guihua Liu , Zixin Chen , Pengxia Yao , Xiaoli Gao
Punicalagin (PUN) is a high abundant ellagitannin and the principal bioactive component existing in Pomegranate (Punica granatum L.) peel, juice, and extract, yet its absorption, bioavailability and pharmacokinetic parameters have not been adequately investigated after oral administration in vivo model. Thus, this study developed a reproducible UPLC-MS/MS method (linear range: 0.125–70 μg/mL; LLOQ: 0.125 μg/mL) to characterize PUN pharmacokinetic profile in Sprague-Dawley rats. After intravenous (iv, 10 mg/kg) administration, PUN exhibited slow elimination (iv, t1/2 = 6.45 ± 2.11 h) and plasma-restricted distribution (Vd = 0.94 ± 0.23 L/kg). By intragastric route (ig, 100–400 mg/kg), PUN showed dose-dependent absorption (Tmax ≈ 2 h), critically low absolute bioavailability (3.22–5.38%) and extensive tissue distribution (Vd = 14.0–44.5 L/kg). The area under the plasma concentration-time curve from time zero to last sampling time (AUC0-t) and AUC0-∞ (from time zero to infinity) were 30.0–211.5, 32.0–213.8 μg*h/mL. The Cmax of PUN in plasma samples was ranged from 1.91 to 34.8 μg/mL. The dose proportionality study demonstrated the Cmax and AUC0-t values were positively correlated with ig doses, with R2 (95% CI) being 0.858 (0.770–0.959) for Cmax and 0.904 (0.847–0.956). Overall, the comprehensively detailed pharmacokinetic parameters of pure PUN have been determined, providing valuable information for preclinical study.
{"title":"A pharmacokinetic study on punicalagin following oral and intravenous administration to the rat using UPLC-MS/MS","authors":"Zhanying Chang , Guihua Liu , Zixin Chen , Pengxia Yao , Xiaoli Gao","doi":"10.1016/j.jchromb.2025.124837","DOIUrl":"10.1016/j.jchromb.2025.124837","url":null,"abstract":"<div><div>Punicalagin (PUN) is a high abundant ellagitannin and the principal bioactive component existing in Pomegranate (<em>Punica granatum</em> L.) peel, juice, and extract, yet its absorption, bioavailability and pharmacokinetic parameters have not been adequately investigated after oral administration <em>in vivo</em> model. Thus, this study developed a reproducible UPLC-MS/MS method (linear range: 0.125–70 μg/mL; LLOQ: 0.125 μg/mL) to characterize PUN pharmacokinetic profile in Sprague-Dawley rats. After intravenous (iv, 10 mg/kg) administration, PUN exhibited slow elimination (iv, t<sub>1/2</sub> = 6.45 ± 2.11 h) and plasma-restricted distribution (V<sub>d</sub> = 0.94 ± 0.23 L/kg). By intragastric route (ig, 100–400 mg/kg), PUN showed dose-dependent absorption (T<sub>max</sub> ≈ 2 h), critically low absolute bioavailability (3.22–5.38%) and extensive tissue distribution (V<sub>d</sub> = 14.0–44.5 L/kg). The area under the plasma concentration-time curve from time zero to last sampling time (AUC<sub>0-t</sub>) and AUC<sub>0-∞</sub> (from time zero to infinity) were 30.0–211.5, 32.0–213.8 μg*h/mL. The C<sub>max</sub> of PUN in plasma samples was ranged from 1.91 to 34.8 μg/mL. The dose proportionality study demonstrated the C<sub>max</sub> and AUC<sub>0-t</sub> values were positively correlated with ig doses, with R<sup>2</sup> (95% CI) being 0.858 (0.770–0.959) for C<sub>max</sub> and 0.904 (0.847–0.956). Overall, the comprehensively detailed pharmacokinetic parameters of pure PUN have been determined, providing valuable information for preclinical study.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124837"},"PeriodicalIF":2.8,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145412459","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}
Pub Date : 2025-10-26DOI: 10.1016/j.jchromb.2025.124838
Maksim Dolov , Igor Rodin
A fast and highly sensitive LC–MS/MS method has been developed and validated for measuring rivastigmine levels in human plasma. The extraction of rivastigmine from plasma was performed using a simple and quick protein precipitation technique (PPT). The internal standard used was atazanavir-d5. Chromatographic separation was achieved using a YMC-Triart C18 column, followed by detection with mass spectrometry. The mass transitions monitored were m/z 251.1 > 206.0 for rivastigmine, and m/z 710.5 > 168.1 for atazanavir-d5. This method involves rapid plasma extraction, straightforward gradient chromatography, and mass spectrometric detection, allowing for the detection of rivastigmine at sub-nanogram per milliliter levels. The method was validated over a linear range of 25 to 5000 pg/ml, with a correlation coefficient of at least 0.9980. Both intra- and inter-day precision and accuracy were within 15 %. The overall recovery rates for rivastigmine and atazanavir-d5 were close to 100 %. The total analysis time per sample was only 3 min. This method was successfully applied to determine the pharmacokinetic parameters of rivastigmine after a single oral dose of 1.5 mg (capsules) in 26 healthy volunteers.
{"title":"Validation of a highly sensitive assay for the determination of rivastigmine in human plasma for pharmacokinetic studies.","authors":"Maksim Dolov , Igor Rodin","doi":"10.1016/j.jchromb.2025.124838","DOIUrl":"10.1016/j.jchromb.2025.124838","url":null,"abstract":"<div><div>A fast and highly sensitive LC–MS/MS method has been developed and validated for measuring rivastigmine levels in human plasma. The extraction of rivastigmine from plasma was performed using a simple and quick protein precipitation technique (PPT). The internal standard used was atazanavir-d<sub>5</sub>. Chromatographic separation was achieved using a YMC-Triart C18 column, followed by detection with mass spectrometry. The mass transitions monitored were <em>m</em>/<em>z</em> 251.1 > 206.0 for rivastigmine, and m/z 710.5 > 168.1 for atazanavir-d<sub>5</sub>. This method involves rapid plasma extraction, straightforward gradient chromatography, and mass spectrometric detection, allowing for the detection of rivastigmine at sub-nanogram per milliliter levels. The method was validated over a linear range of 25 to 5000 pg/ml, with a correlation coefficient of at least 0.9980. Both intra- and inter-day precision and accuracy were within 15 %. The overall recovery rates for rivastigmine and atazanavir-d<sub>5</sub> were close to 100 %. The total analysis time per sample was only 3 min. This method was successfully applied to determine the pharmacokinetic parameters of rivastigmine after a single oral dose of 1.5 mg (capsules) in 26 healthy volunteers.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124838"},"PeriodicalIF":2.8,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145415412","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}
Pub Date : 2025-10-26DOI: 10.1016/j.jchromb.2025.124836
Hae-In Choi , Hyeon-Cheol Jeong , Jong-Woo Jeong , Jaeyoung Lee , Da Hae Kim , Kyong-Cheol Ko , Yoon-Jee Chae , Kyeong-Ryoon Lee
A sensitive and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of tirzepatide, a dual agonist of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors in rat plasma. Tirzepatide was extracted from rat plasma by protein precipitation using methanol. Chromatographic separation was achieved using a peptide C18 column with gradient elution of water and acetonitrile containing 0.1 % formic acid. Mass spectrometric detection was performed in positive electrospray ionization mode using multiple reaction monitoring with transitions of m/z 1204.4 → 1473.6 for tirzepatide and m/z 1029.4 → 1238.4 for the internal standard, semaglutide. The developed method exhibited good linearity over a concentration range of 1–1000 ng/mL (r2 > 0.99). Intra- and inter-day accuracy (−4.324–5.057 %) and precision (5.250–9.000 %) met the regulatory criteria at all quality control levels, and were stable under various plasma handling and storage conditions. The validated method was successfully applied to a pharmacokinetic study in rats following the intravenous and subcutaneous injection of tirzepatide at 0.3 mg/kg. The terminal half-lives were 10.04 h and 9.803 h after intravenous and subcutaneous administration, respectively, indicating comparable elimination profiles. The bioavailability following subcutaneous dosing was estimated to be approximately 62.38 %. These findings highlight the robustness and applicability of the developed method, suggesting its potential utility for the quantitative analysis of other peptide therapeutics with structures or mechanisms of action similar to those of tirzepatide.
{"title":"Development and validation of an LC-MS/MS method for Tirzepatide, a dual GIP/GLP-1 receptor agonist, in rat plasma for application to a pharmacokinetic study","authors":"Hae-In Choi , Hyeon-Cheol Jeong , Jong-Woo Jeong , Jaeyoung Lee , Da Hae Kim , Kyong-Cheol Ko , Yoon-Jee Chae , Kyeong-Ryoon Lee","doi":"10.1016/j.jchromb.2025.124836","DOIUrl":"10.1016/j.jchromb.2025.124836","url":null,"abstract":"<div><div>A sensitive and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of tirzepatide, a dual agonist of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors in rat plasma. Tirzepatide was extracted from rat plasma by protein precipitation using methanol. Chromatographic separation was achieved using a peptide C18 column with gradient elution of water and acetonitrile containing 0.1 % formic acid. Mass spectrometric detection was performed in positive electrospray ionization mode using multiple reaction monitoring with transitions of <em>m</em>/<em>z</em> 1204.4 → 1473.6 for tirzepatide and m/z 1029.4 → 1238.4 for the internal standard, semaglutide. The developed method exhibited good linearity over a concentration range of 1–1000 ng/mL (r<sup>2</sup> > 0.99). Intra- and inter-day accuracy (−4.324–5.057 %) and precision (5.250–9.000 %) met the regulatory criteria at all quality control levels, and were stable under various plasma handling and storage conditions. The validated method was successfully applied to a pharmacokinetic study in rats following the intravenous and subcutaneous injection of tirzepatide at 0.3 mg/kg. The terminal half-lives were 10.04 h and 9.803 h after intravenous and subcutaneous administration, respectively, indicating comparable elimination profiles. The bioavailability following subcutaneous dosing was estimated to be approximately 62.38 %. These findings highlight the robustness and applicability of the developed method, suggesting its potential utility for the quantitative analysis of other peptide therapeutics with structures or mechanisms of action similar to those of tirzepatide.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1268 ","pages":"Article 124836"},"PeriodicalIF":2.8,"publicationDate":"2025-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145461102","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}
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