Journal of pharmaceutical and biomedical analysis最新文献_第2页

Determination of Gentamicin C-subtypes in Inner Ear Perilymph Using Liquid Chromatography with Fluorescence Detection 液相色谱-荧光检测法测定内耳淋巴周围庆大霉素c亚型。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-05 DOI: 10.1016/j.jpba.2026.117394

Shreshtha Dash, Molly T. McDevitt, D. David Smith, Peter S. Steyger

Gentamicin is a broad-spectrum aminoglycoside used frequently to treat gram-positive and gram-negative bacterial infections. In this study, a new, simple, fast, and sensitive isocratic reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the detection and quantification of fluorescent OPA-ethanethiol derivatized gentamicin in very small biological sample volumes. To our knowledge, there is no report of the use of ethanethiol for the derivatization of gentamicin with OPA, and the simultaneous determination of the four major C-subtypes of gentamicin using OPA-derivatives. Optimum chromatographic conditions were achieved on a C₁₈ column with a mobile phase consisting of methanol, glacial acetic acid, and an aqueous solution of sodium 1-heptanesulfonate at a flow rate of 1.0 mL/min under ambient conditions. The method was successfully validated according to the acceptance criteria of USP guidelines in terms of selectivity, linearity, accuracy, precision, and sensitivity. The linearity of the method was demonstrated with a concentration range of gentamicin (10-400 ng/mL) prepared in artificial perilymph. The limit of detection was 0.2 ng/mL and the limit of quantification was 10–11 ng/mL for all four major C-subtypes of gentamicin. Finally, due to its high sensitivity, this method was successfully applied to quantify gentamicin concentrations in the small volumes of perilymph present in the inner ear of mice. Thus, this RP-HPLC-fluorescence method for detecting derivatized gentamicin in preclinical models is promising in terms of simplicity and high sensitivity.

庆大霉素是一种广谱氨基糖苷，常用于治疗革兰氏阳性和革兰氏阴性细菌感染。本研究建立了一种新的、简单、快速、灵敏的等容反相高效液相色谱（RP-HPLC）方法，用于极小生物样品体积下opa -乙硫醇衍生庆大霉素的荧光检测和定量。据我们所知，目前还没有使用乙硫醇与OPA衍生庆大霉素的报道，也没有使用OPA衍生物同时测定庆大霉素的四个主要c亚型的报道。在C18色谱柱上，以甲醇、冰醋酸和1-庚烷磺酸钠水溶液为流动相，流速为1.0 mL/min，获得了最佳色谱条件。方法在选择性、线性度、准确度、精密度、灵敏度等方面均符合美国药典的验收标准。在人工淋巴周围制备的庆大霉素（10 ~ 400 ng/mL）浓度范围内，本法线性良好。4种主要c亚型庆大霉素的检测限为0.2 ng/mL，定量限为10-11 ng/mL。最后，由于该方法灵敏度高，成功地应用于小鼠内耳淋巴周围小体积庆大霉素浓度的定量。因此，这种rp - hplc -荧光法在临床前模型中检测衍生庆大霉素具有简便和高灵敏度的优点。

{"title":"Determination of Gentamicin C-subtypes in Inner Ear Perilymph Using Liquid Chromatography with Fluorescence Detection","authors":"Shreshtha Dash, Molly T. McDevitt, D. David Smith, Peter S. Steyger","doi":"10.1016/j.jpba.2026.117394","DOIUrl":"10.1016/j.jpba.2026.117394","url":null,"abstract":"<div><div>Gentamicin is a broad-spectrum aminoglycoside used frequently to treat gram-positive and gram-negative bacterial infections. In this study, a new, simple, fast, and sensitive isocratic reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the detection and quantification of fluorescent OPA-ethanethiol derivatized gentamicin in very small biological sample volumes. To our knowledge, there is no report of the use of ethanethiol for the derivatization of gentamicin with OPA, and the simultaneous determination of the four major C-subtypes of gentamicin using OPA-derivatives. Optimum chromatographic conditions were achieved on a C<sub>18</sub> column with a mobile phase consisting of methanol, glacial acetic acid, and an aqueous solution of sodium 1-heptanesulfonate at a flow rate of 1.0 mL/min under ambient conditions. The method was successfully validated according to the acceptance criteria of USP guidelines in terms of selectivity, linearity, accuracy, precision, and sensitivity. The linearity of the method was demonstrated with a concentration range of gentamicin (10-400 ng/mL) prepared in artificial perilymph. The limit of detection was 0.2 ng/mL and the limit of quantification was 10–11 ng/mL for all four major C-subtypes of gentamicin. Finally, due to its high sensitivity, this method was successfully applied to quantify gentamicin concentrations in the small volumes of perilymph present in the inner ear of mice. Thus, this RP-HPLC-fluorescence method for detecting derivatized gentamicin in preclinical models is promising in terms of simplicity and high sensitivity.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117394"},"PeriodicalIF":3.1,"publicationDate":"2026-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146157367","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

Aggregation-induced luminescence probe based lateral flow immunoassay for the simultaneous quantitative detection of IL-6/PCT 基于聚集诱导发光探针的侧流免疫法同时定量检测IL-6/PCT。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-05 DOI: 10.1016/j.jpba.2026.117393

Rong He , Yanjie Zhu , Weiping Zhou , Muhammad Azhar Hayat Nawaz , Yunhui Li , Jianwei Zhu , Huimin Feng , Anna Maria Nowicka , Wenzhao Han , Cong Yu

The detection of important inflammatory biomarkers possesses substantial advantages in guiding clinical decision-making. In particular, simultaneous detection of interleukin-6 (IL-6) and procalcitonin (PCT) significantly improves the differentiation between bacterial and viral infections, a critical challenge in early-stage diagnostics. To address this need, a reliable lateral flow immunoassay (LFIA) incorporating aggregation-induced luminescent nanoparticles (BTA@PS) was successfully developed. A novel aggregation-induced emission (AIE) probe was designed and encapsulated within polystyrene microspheres, thereby overcoming the limitations of aggregation-caused quenching (ACQ) in conventional fluorescent materials. The resulting BTA@PS nanoparticles, conjugated with specific antibodies against IL-6 and PCT, served as stable and effective immunofluorescent probes for the LFIA platform. Under optimized experimental conditions, the developed BTA@PS-LFIA enabled simultaneous quantification of IL-6 and PCT, demonstrating excellent linearity over the range of 2–8000 pg/mL and 0.04–30 ng/mL, respectively, with coefficient of variation (CV) values of below 5 %. Furthermore, this method demonstrated superior detection capability for PCT and IL-6 in serum, confirming its high potential for rapid clinical diagnostics.

重要炎症生物标志物的检测在指导临床决策方面具有实质性的优势。特别是，同时检测白细胞介素-6 （IL-6）和降钙素原（PCT）可显著提高细菌和病毒感染的区分，这是早期诊断的一个关键挑战。为了满足这一需求，成功开发了一种可靠的横向流动免疫测定（LFIA），其中包含聚集诱导的发光纳米颗粒（BTA@PS）。设计了一种聚苯乙烯微球封装的新型聚集致猝灭（AIE）探针，克服了传统荧光材料中聚集致猝灭（ACQ）的局限性。所得BTA@PS纳米颗粒结合了针对IL-6和PCT的特异性抗体，作为LFIA平台稳定有效的免疫荧光探针。在优化的实验条件下，建立的BTA@PS-LFIA可以同时定量IL-6和PCT，分别在2-8000 pg/mL和0.04-30 ng/mL范围内具有良好的线性关系，变异系数（CV）值小于5 %。此外，该方法对血清中PCT和IL-6的检测能力优越，证实了其在快速临床诊断中的巨大潜力。

{"title":"Aggregation-induced luminescence probe based lateral flow immunoassay for the simultaneous quantitative detection of IL-6/PCT","authors":"Rong He , Yanjie Zhu , Weiping Zhou , Muhammad Azhar Hayat Nawaz , Yunhui Li , Jianwei Zhu , Huimin Feng , Anna Maria Nowicka , Wenzhao Han , Cong Yu","doi":"10.1016/j.jpba.2026.117393","DOIUrl":"10.1016/j.jpba.2026.117393","url":null,"abstract":"<div><div>The detection of important inflammatory biomarkers possesses substantial advantages in guiding clinical decision-making. In particular, simultaneous detection of interleukin-6 (IL-6) and procalcitonin (PCT) significantly improves the differentiation between bacterial and viral infections, a critical challenge in early-stage diagnostics. To address this need, a reliable lateral flow immunoassay (LFIA) incorporating aggregation-induced luminescent nanoparticles (BTA@PS) was successfully developed. A novel aggregation-induced emission (AIE) probe was designed and encapsulated within polystyrene microspheres, thereby overcoming the limitations of aggregation-caused quenching (ACQ) in conventional fluorescent materials. The resulting BTA@PS nanoparticles, conjugated with specific antibodies against IL-6 and PCT, served as stable and effective immunofluorescent probes for the LFIA platform. Under optimized experimental conditions, the developed BTA@PS-LFIA enabled simultaneous quantification of IL-6 and PCT, demonstrating excellent linearity over the range of 2–8000 pg/mL and 0.04–30 ng/mL, respectively, with coefficient of variation (CV) values of below 5 %. Furthermore, this method demonstrated superior detection capability for PCT and IL-6 in serum, confirming its high potential for rapid clinical diagnostics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117393"},"PeriodicalIF":3.1,"publicationDate":"2026-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146180304","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

Temperature as the primary risk factor for sevoflurane degradation: Identification and toxicological risk assessment of novel degradants by nuclear magnetic resonance (NMR) spectroscopy 温度是七氟醚降解的主要危险因素：核磁共振（NMR）光谱技术鉴定新型降解剂及毒理学风险评估。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-04 DOI: 10.1016/j.jpba.2026.117391

Yi Jin , Yaning Li , Haoran Yang , Huiying Yang , Yaqing Guo , Xianfu Wu

Sevoflurane is an inhalation anesthetic, which is widely favored by anesthesiologists in modern clinical practice. It is reported that sevoflurane readily degrades under certain conditions. Forced degradation studies are essential for elucidating its degradation profile under stressed conditions, as recommended by the ICH guidelines. This study systematically assessed the impact of temperature, light, oxidation, and hydrolysis on sevoflurane stability through forced degradation studies. The results indicated that temperature was identified as the primary trigger for degradation, leading to the formation of four degradation products (DPs) (including the pharmacopeial-known Impurity C and three novel sevoflurane degradants H, I, and J) at 60℃ and 95℃. Light, oxidation, and hydrolysis had no significant effect under the tested conditions. The structures of all DPs were confirmed using nuclear magnetic resonance spectroscopy (NMR). Capitalizing on the inherent quantitative proficiency of this technique, we subsequently employed quantitative proton nuclear magnetic resonance spectroscopy (qHNMR) to determine the relative contents of these four DPs, which can further elucidate the degradation profile of sevoflurane. In silico toxicity and metabolic behavior of them were assessed by Derek Nexus and Meteor Nexus software, respectively. DPs H-J exhibited potential skin irritation/corrosion and sensitization effects across species, potentially attributable to alkyl aldehyde functional groups. All of our efforts are expected to provide guidance for the quality control and optimal storage of sevoflurane.

七氟醚是一种吸入性麻醉剂，在现代临床实践中受到麻醉医师的广泛青睐。据报道，七氟醚在一定条件下容易降解。根据ICH指南的建议，强制降解研究对于阐明其在压力条件下的降解概况至关重要。本研究通过强制降解研究系统地评估了温度、光照、氧化和水解对七氟醚稳定性的影响。结果表明，温度是降解的主要触发因素，在60℃和95℃下形成4种降解产物（包括药典中已知的杂质C和3种新型七氟烷降解物H、I和J）。光照、氧化和水解在测试条件下没有显著影响。所有DPs的结构都用核磁共振波谱（NMR）证实。利用该技术固有的定量能力，我们随后采用定量质子核磁共振波谱（qHNMR）测定了这四种DPs的相对含量，这可以进一步阐明七氟醚的降解谱。采用Derek Nexus和Meteor Nexus软件分别评价其硅毒性和代谢行为。DPs H-J在不同物种间表现出潜在的皮肤刺激/腐蚀和致敏效应，可能归因于烷基醛官能团。我们的所有努力都有望为七氟醚的质量控制和最佳储存提供指导。

{"title":"Temperature as the primary risk factor for sevoflurane degradation: Identification and toxicological risk assessment of novel degradants by nuclear magnetic resonance (NMR) spectroscopy","authors":"Yi Jin , Yaning Li , Haoran Yang , Huiying Yang , Yaqing Guo , Xianfu Wu","doi":"10.1016/j.jpba.2026.117391","DOIUrl":"10.1016/j.jpba.2026.117391","url":null,"abstract":"<div><div>Sevoflurane is an inhalation anesthetic, which is widely favored by anesthesiologists in modern clinical practice. It is reported that sevoflurane readily degrades under certain conditions. Forced degradation studies are essential for elucidating its degradation profile under stressed conditions, as recommended by the ICH guidelines. This study systematically assessed the impact of temperature, light, oxidation, and hydrolysis on sevoflurane stability through forced degradation studies. The results indicated that temperature was identified as the primary trigger for degradation, leading to the formation of four degradation products (DPs) (including the pharmacopeial-known Impurity C and three novel sevoflurane degradants H, I, and J) at 60℃ and 95℃. Light, oxidation, and hydrolysis had no significant effect under the tested conditions. The structures of all DPs were confirmed using nuclear magnetic resonance spectroscopy (NMR). Capitalizing on the inherent quantitative proficiency of this technique, we subsequently employed quantitative proton nuclear magnetic resonance spectroscopy (qHNMR) to determine the relative contents of these four DPs, which can further elucidate the degradation profile of sevoflurane. <em>In silico</em> toxicity and metabolic behavior of them were assessed by Derek Nexus and Meteor Nexus software, respectively. DPs H-J exhibited potential skin irritation/corrosion and sensitization effects across species, potentially attributable to alkyl aldehyde functional groups. All of our efforts are expected to provide guidance for the quality control and optimal storage of sevoflurane.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117391"},"PeriodicalIF":3.1,"publicationDate":"2026-02-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146157363","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

In memoriam of Professor Sándor Görög 为纪念Sándor Görög教授。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-04 DOI: 10.1016/j.jpba.2026.117395

引用次数: 0

Determination of antigen components in inactivated SARS-CoV-2 vaccine using ultra-high-performance liquid chromatography tandem mass spectrometry 超高效液相色谱串联质谱法测定SARS-CoV-2灭活疫苗抗原成分

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-02 DOI: 10.1016/j.jpba.2026.117388

Jiexia Shi , Fenfang Deng , Yongxian Li , Juntao Li , Rongfei Peng , Jun Yuan , Lei Tan

Accurate and reliable quantification of antigen components in vaccines is critical in vaccine quality control and evaluation of immunogenic consistency. However, conventional immunoassays often suffer from limited specificity, trace-level antigen concentrations, and indirect quantification. In this study, we demonstrated an ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the determination of effective antigen components in inactivated SARS-CoV-2 vaccines. Specifically, the vaccine samples were denatured and digested with trypsin to generate tryptic peptides. Then, the signature peptides derived from the nucleocapsid protein and their stable isotope–labeled internal standards were selectively captured and separated using anti-peptide antibody-conjugated magnetic beads. The signature peptides were characterized by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, which confirmed their amino acid sequence and multi-charged ionization states. Quantitative analysis was then performed using UHPLC-MS/MS in positive electrospray ionization mode with multiple reaction monitoring. Chromatographic separation of the signature peptides was achieved on an ACQUITY Premier Peptide BEH C₁₈ column using 0.1 % formic acid in water and 0.1 % formic acid in acetonitrile as the mobile phases. The method was validated and exhibited excellent linearity for the signature peptides over the concentration range of 1–60 μg/L, with correlation coefficients higher than 0.999. The recovery ranged from 75.1 % to 86.3 %, with intra-day precision (RSD) of 0.8–1.0 % and inter-day precision of 1.3–3.6 %. Finally, the method was successfully applied to determine the effective antigen components in inactivated SARS-CoV-2 vaccine samples. The concentrations of the signature peptide ADETQALPQR ranged from 4.95 to 12.95 µg/L across the three vaccine batches analyzed, corresponding to 4.38–11.47 nmol/L of nucleocapsid protein. The results indicated that the method exhibited great promise for the determination of active antigenic proteins in inactivated SARS-CoV-2 vaccine samples and provided an alternative analytical platform for vaccine quality control.

疫苗中抗原成分的准确和可靠定量对疫苗质量控制和免疫原性一致性评价至关重要。然而，传统的免疫测定法往往具有有限的特异性、微量水平的抗原浓度和间接定量。本研究建立了一种超高效液相色谱-串联质谱（UHPLC-MS/MS）测定SARS-CoV-2灭活疫苗中有效抗原成分的方法。具体地说，疫苗样品变性和胰蛋白酶消化产生胰蛋白酶肽。然后，利用抗肽抗体偶联磁珠选择性捕获核衣壳蛋白的特征肽及其稳定的同位素标记内标。采用超高效液相色谱-四极杆飞行时间质谱法对特征肽进行了表征，确定了它们的氨基酸序列和多电荷电离态。采用超高效液相色谱-质谱联用（UHPLC-MS/MS）进行定量分析，电喷雾电离模式为多反应监测。特征肽的色谱分离在ACQUITY Premier Peptide BEH C₁₈色谱柱上实现，流动相为0.1 %甲酸水溶液和0.1 %甲酸乙腈水溶液。该方法在1 ~ 60 μg/L范围内具有良好的线性关系，相关系数大于0.999。加样回收率为75.1% % ~ 86.3% %，日内精密度（RSD）为0.8 ~ 1.0 %，日内精密度为1.3 ~ 3.6 %。最后，将该方法成功应用于SARS-CoV-2灭活疫苗样品中有效抗原组分的测定。特征肽addetqalpqr在三个疫苗批次中的浓度范围为4.95 ~ 12.95 µg/L，对应于4.38 ~ 11.47 nmol/L的核衣壳蛋白。结果表明，该方法可用于灭活SARS-CoV-2疫苗样品中活性抗原蛋白的测定，为疫苗质量控制提供了新的分析平台。

{"title":"Determination of antigen components in inactivated SARS-CoV-2 vaccine using ultra-high-performance liquid chromatography tandem mass spectrometry","authors":"Jiexia Shi , Fenfang Deng , Yongxian Li , Juntao Li , Rongfei Peng , Jun Yuan , Lei Tan","doi":"10.1016/j.jpba.2026.117388","DOIUrl":"10.1016/j.jpba.2026.117388","url":null,"abstract":"<div><div>Accurate and reliable quantification of antigen components in vaccines is critical in vaccine quality control and evaluation of immunogenic consistency. However, conventional immunoassays often suffer from limited specificity, trace-level antigen concentrations, and indirect quantification. In this study, we demonstrated an ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the determination of effective antigen components in inactivated SARS-CoV-2 vaccines. Specifically, the vaccine samples were denatured and digested with trypsin to generate tryptic peptides. Then, the signature peptides derived from the nucleocapsid protein and their stable isotope–labeled internal standards were selectively captured and separated using anti-peptide antibody-conjugated magnetic beads. The signature peptides were characterized by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, which confirmed their amino acid sequence and multi-charged ionization states. Quantitative analysis was then performed using UHPLC-MS/MS in positive electrospray ionization mode with multiple reaction monitoring. Chromatographic separation of the signature peptides was achieved on an ACQUITY Premier Peptide BEH C₁₈ column using 0.1 % formic acid in water and 0.1 % formic acid in acetonitrile as the mobile phases. The method was validated and exhibited excellent linearity for the signature peptides over the concentration range of 1–60 μg/L, with correlation coefficients higher than 0.999. The recovery ranged from 75.1 % to 86.3 %, with intra-day precision (RSD) of 0.8–1.0 % and inter-day precision of 1.3–3.6 %. Finally, the method was successfully applied to determine the effective antigen components in inactivated SARS-CoV-2 vaccine samples. The concentrations of the signature peptide ADETQALPQR ranged from 4.95 to 12.95 µg/L across the three vaccine batches analyzed, corresponding to 4.38–11.47 nmol/L of nucleocapsid protein. The results indicated that the method exhibited great promise for the determination of active antigenic proteins in inactivated SARS-CoV-2 vaccine samples and provided an alternative analytical platform for vaccine quality control.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117388"},"PeriodicalIF":3.1,"publicationDate":"2026-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146125450","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 and validation of a sensitive and rapid UHPLC-MS/MS method for the simultaneous quantification of CG-0255 and its active metabolite in human plasma and its application to Phase I studies 高效液相色谱-质谱联用同时定量人血浆中CG-0255及其活性代谢物的方法的建立与验证及其在I期研究中的应用

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-02 DOI: 10.1016/j.jpba.2026.117390

Hanjing Chen , Jiali Li , Fei Yuan , Gongxin He , Hao Wu , Hua Yan , Hongrong Xu , Chao Liu , Lei Sheng , Xuening Li

CG-0255, a thiol prodrug of clopidogrel’s active metabolite H4 (CG-0236), is a novel thienopyridine P2Y12 receptor antagonist under initial clinical development for the treatment of acute coronary syndromes. Unlike clopidogrel, CG-0255 is converted to the active thiol metabolite H4 (CG-0236) in a single hydrolytic step. Compared with clopidogrel, CG-0255 exhibits more efficient and consistent H4 formation in humans, which can be quantified in plasma following either intravenous or oral administration. In this study, we developed and validated a sensitive, rapid, and robust UHPLC–MS/MS method for the simultaneous quantification of CG-0255 and its derivatized active metabolite (MP-H4, CG-0261) in human plasma. After solid-phase extraction from 94.5 μL of plasma, analytes and isotope-labeled internal standards were separated on an ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using isocratic elution with 0.1 % formic acid in water and acetonitrile (57:43, v/v) at a flow rate of 0.5 mL/min, followed by a 3.5 min column washing and re-equilibration, giving a total analytical run time of 7 min. Baseline separation of CG-0255, CG-0261, and their respective isomers was achieved. Detection was performed using positive electrospray ionization in multiple reaction monitoring mode on a Q-Trap 6500⁺ mass spectrometer. Calibration curves were linear over 0.05–25 ng/mL for both analytes, corresponding to 0.0353–17.65 ng/mL for H4 (CG-0236). Intra- and inter-day precision and accuracy were within ±15 % at all quality-control levels. The validated assay was successfully applied to two phase I clinical studies conducted at our center, characterizing the pharmacokinetics of CG-0255 following single-dose intravenous and multiple-dose oral administration. This UHPLC–MS/MS method provides a reliable platform for the quantitative evaluation of CG-0255 and its active metabolite in human plasma, and is well suited to support further global clinical development.

CG-0255是氯吡格雷活性代谢物H4 （CG-0236）的巯基前药，是一种新型噻吩吡啶P2Y12受体拮抗剂，正在初步临床开发中，用于治疗急性冠状动脉综合征。与氯吡格雷不同，CG-0255在一个水解步骤中转化为活性硫醇代谢物H4 （CG-0236）。与氯吡格雷相比，CG-0255在人体内表现出更有效和一致的H4形成，可以在静脉或口服给药后在血浆中量化。在本研究中，我们建立并验证了一种灵敏、快速、稳健的UHPLC-MS/MS同时定量人血浆中CG-0255及其衍生活性代谢物（MP-H4, CG-0261）的方法。固相萃取94.5 μL血浆后，用ACQUITY UPLC BEH C18色谱柱（2.1 mm × 50 mm, 1.7 μm）分离分析物和同位素标记的内标物，用0.1 %甲酸水溶液和乙腈（57:43,v/v）等压洗脱，流速0.5 mL/min，洗涤3.5 min，再平衡，总分析运行时间7 min。对CG-0255、CG-0261及其异构体进行了基线分离。在Q-Trap 6500+质谱仪上采用多反应监测模式下的正电喷雾电离进行检测。两种分析物的校准曲线在0.05-25 ng/mL范围内呈线性，对应于H4 （CG-0236）的校准曲线为0.0353-17.65 ng/mL。在所有质量控制水平下，日内、日间精密度和准确度均在±15 %以内。验证的检测方法已成功应用于我们中心进行的两项I期临床研究，表征了CG-0255单剂量静脉注射和多剂量口服给药后的药代动力学。该UHPLC-MS/MS方法为CG-0255及其在人血浆中的活性代谢物的定量评价提供了可靠的平台，非常适合支持进一步的全球临床开发。

{"title":"Development and validation of a sensitive and rapid UHPLC-MS/MS method for the simultaneous quantification of CG-0255 and its active metabolite in human plasma and its application to Phase I studies","authors":"Hanjing Chen , Jiali Li , Fei Yuan , Gongxin He , Hao Wu , Hua Yan , Hongrong Xu , Chao Liu , Lei Sheng , Xuening Li","doi":"10.1016/j.jpba.2026.117390","DOIUrl":"10.1016/j.jpba.2026.117390","url":null,"abstract":"<div><div>CG-0255, a thiol prodrug of clopidogrel’s active metabolite H4 (CG-0236), is a novel thienopyridine P2Y12 receptor antagonist under initial clinical development for the treatment of acute coronary syndromes. Unlike clopidogrel, CG-0255 is converted to the active thiol metabolite H4 (CG-0236) in a single hydrolytic step. Compared with clopidogrel, CG-0255 exhibits more efficient and consistent H4 formation in humans, which can be quantified in plasma following either intravenous or oral administration. In this study, we developed and validated a sensitive, rapid, and robust UHPLC–MS/MS method for the simultaneous quantification of CG-0255 and its derivatized active metabolite (MP-H4, CG-0261) in human plasma. After solid-phase extraction from 94.5 μL of plasma, analytes and isotope-labeled internal standards were separated on an ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using isocratic elution with 0.1 % formic acid in water and acetonitrile (57:43, v/v) at a flow rate of 0.5 mL/min, followed by a 3.5 min column washing and re-equilibration, giving a total analytical run time of 7 min. Baseline separation of CG-0255, CG-0261, and their respective isomers was achieved. Detection was performed using positive electrospray ionization in multiple reaction monitoring mode on a Q-Trap 6500<sup>+</sup> mass spectrometer. Calibration curves were linear over 0.05–25 ng/mL for both analytes, corresponding to 0.0353–17.65 ng/mL for H4 (CG-0236). Intra- and inter-day precision and accuracy were within ±15 % at all quality-control levels. The validated assay was successfully applied to two phase I clinical studies conducted at our center, characterizing the pharmacokinetics of CG-0255 following single-dose intravenous and multiple-dose oral administration. This UHPLC–MS/MS method provides a reliable platform for the quantitative evaluation of CG-0255 and its active metabolite in human plasma, and is well suited to support further global clinical development.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117390"},"PeriodicalIF":3.1,"publicationDate":"2026-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146132099","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

Corrigendum to ‘A liquid biopsy-RNAseq method for monitoring the expression of genes involved in drug disposition: proof-of-concept application to cholestatic liver disease’ [J. Pharm. Biomed. Anal. (2025) 269: 117244] “液体活检- rnaseq方法监测药物处置相关基因表达：概念验证应用于胆汁淤积性肝病”的更正[J]。制药。生物医学。分析的。（2025） 269: 117244]

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-02 DOI: 10.1016/j.jpba.2026.117385

Amit Dahal , Teresa Sierra , Colleen M. Hayes , David N. Assis , Amin Rostami-Hodjegan , Nisanne S. Ghonem , Brahim Achour

引用次数: 0

Development and validation of an UPLC–MS/MS method for simultaneous determination of meropenem and its open-ring metabolite in human serum and cerebrospinal fluid with application to clinical samples 同时测定人血清和脑脊液中美罗培南及其开环代谢物的UPLC-MS/MS方法的建立及应用于临床样品的验证

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-02 DOI: 10.1016/j.jpba.2026.117389

XiangLong Chen , Jinhui Xu , Chengliang Wang, Lijuan Yang, Jinwei Fan, Tongtong Li, Qian Zhang, Yanxia Yu, Lian Tang, Shenjia Huang

Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradation to an inactive open-ring metabolite (ORM). In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of MER and ORM in human serum and CSF. Chromatographic separation was accomplished on an Agela Venusil MP C18 column, with MER-d6 and ORM-d6 as internal standards. Methanol was found to promote methanolysis, yielding a characteristic product (m/z 416.2). Therefore, acetonitrile was selected as both the organic phase and the protein-precipitation solvent. Method validation was conducted according to the ICH M10 guideline. Follow validation, the method was successfully applied to 57 serum and 16 CSF samples. ORM concentrations in human CSF were reported for the first time. This method provides a valuable tool to support MER monitoring in patients with CNS infections.

中枢神经系统（CNS）感染需要在作用部位充分暴露于药物，但抗生素美罗苯南（MER）显示脑脊液（CSF）渗透有限，并且容易经历非酶降解为无活性开环代谢物（ORM）。本研究建立了一种简便、灵敏、快速的液相色谱串联质谱（LC-MS/MS）同时测定人血清和脑脊液中MER和ORM的方法。色谱分离采用Agela Venusil MP C18色谱柱，MER-d6和ORM-d6为内标。发现甲醇促进甲醇分解，产生特征产物（m/z 416.2）。因此，选择乙腈作为有机相和蛋白质沉淀溶剂。方法验证按照ICH M10指南进行。经过验证，该方法成功应用于57份血清和16份脑脊液样本。ORM在人脑脊液中的浓度为首次报道。该方法为支持中枢神经系统感染患者的MER监测提供了有价值的工具。

{"title":"Development and validation of an UPLC–MS/MS method for simultaneous determination of meropenem and its open-ring metabolite in human serum and cerebrospinal fluid with application to clinical samples","authors":"XiangLong Chen , Jinhui Xu , Chengliang Wang, Lijuan Yang, Jinwei Fan, Tongtong Li, Qian Zhang, Yanxia Yu, Lian Tang, Shenjia Huang","doi":"10.1016/j.jpba.2026.117389","DOIUrl":"10.1016/j.jpba.2026.117389","url":null,"abstract":"<div><div>Central nervous system (CNS) infections require adequate drug exposure at the site of action, yet antibiotic meropenem (MER) shows limited cerebrospinal fluid (CSF) penetration and easily undergoes non-enzymatic degradation to an inactive open-ring metabolite (ORM). In this study, we developed a simple, sensitive, and fast liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of MER and ORM in human serum and CSF. Chromatographic separation was accomplished on an Agela Venusil MP C18 column, with MER-d6 and ORM-d6 as internal standards. Methanol was found to promote methanolysis, yielding a characteristic product (<em>m/z</em> 416.2). Therefore, acetonitrile was selected as both the organic phase and the protein-precipitation solvent. Method validation was conducted according to the ICH M10 guideline. Follow validation, the method was successfully applied to 57 serum and 16 CSF samples. ORM concentrations in human CSF were reported for the first time. This method provides a valuable tool to support MER monitoring in patients with CNS infections.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117389"},"PeriodicalIF":3.1,"publicationDate":"2026-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119315","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

Fecal metabolic biomarkers associated with insomnia severity: A study on 5-hydroxyindoleacetic acid, octopamine, oleoylethanolamide, and elaidic acid 与失眠严重程度相关的粪便代谢生物标志物：5-羟基吲哚乙酸、章鱼胺、油基乙醇酰胺和elaidi酸的研究

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-02-01 DOI: 10.1016/j.jpba.2026.117387

Yinuo Wu , Xiaoli Li , Haixia Feng , Yifan Chen , Wenwen Wu , Siqin Wang

Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (OEA), and elaidic acid (EA) in patients with sleep disorders, as well as their correlations with insomnia severity. Sixty participants were divided into two groups, with thirty patients with sleep disorders hospitalized in the Department of Neurology, Zhongda Hospital, Southeast University (October 2024–March 2025) and 30 healthy controls recruited during the same period. Fecal samples were collected from all participants, and metabolite levels were analyzed via untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS). The Chinese version of the Insomnia Severity Index (C-ISI) was employed to evaluate insomnia severity, and the correlations between insomnia severity and these four metabolites were subjected to statistical analysis. Both univariate and multivariate analyses revealed significant metabolic differences between groups. The experimental group showed significantly lower levels of 5-HIAA (FC = 0.947, P = 0.020) and OA (FC = 0.953, P < 0.001), but higher OEA (FC = 1.101, P < 0.001) and EA (FC = 1.026, P < 0.001). C-ISI scores correlated negatively with 5-HIAA (r = -0.380, P = 0.003) and OA (r = -0.448, P < 0.001), and positively with OEA (r = 0.500, P < 0.001) and EA (r = 0.408, P = 0.001). These fecal metabolites associate with insomnia severity and may serve as potential biomarkers for understanding its pathophysiology and developing interventions.

人们越来越认识到失眠是一种复杂代谢基础的疾病。我们研究了睡眠障碍患者粪便代谢物5-羟基吲哚乙酸（5-HIAA）、章鱼胺（OA）、油基乙醇酰胺（OEA）和elaidic酸（EA）水平的变化及其与失眠严重程度的相关性。60名受试者分为两组，选取东南大学中大医院神经内科住院的睡眠障碍患者30例（2024年10月- 2025年3月）和同期招募的健康对照30例。收集所有参与者的粪便样本，并通过液相色谱-质谱（LC-MS）非靶向代谢组学分析分析代谢物水平。采用中文版失眠症严重程度指数（C-ISI）评估失眠严重程度，并对失眠严重程度与上述四种代谢物的相关性进行统计分析。单因素和多因素分析均显示各组之间的代谢差异显著。实验组5-HIAA （FC = 0.947, P = 0.020）和OA （FC = 0.953, P . 0.05）水平显著低于对照组

{"title":"Fecal metabolic biomarkers associated with insomnia severity: A study on 5-hydroxyindoleacetic acid, octopamine, oleoylethanolamide, and elaidic acid","authors":"Yinuo Wu , Xiaoli Li , Haixia Feng , Yifan Chen , Wenwen Wu , Siqin Wang","doi":"10.1016/j.jpba.2026.117387","DOIUrl":"10.1016/j.jpba.2026.117387","url":null,"abstract":"<div><div>Insomnia is increasingly recognized as a disorder with complex metabolic underpinnings. We investigated the changes in fecal metabolite levels of 5-hydroxyindoleacetic acid (5-HIAA), octopamine (OA), oleoylethanolamide (OEA), and elaidic acid (EA) in patients with sleep disorders, as well as their correlations with insomnia severity. Sixty participants were divided into two groups, with thirty patients with sleep disorders hospitalized in the Department of Neurology, Zhongda Hospital, Southeast University (October 2024–March 2025) and 30 healthy controls recruited during the same period. Fecal samples were collected from all participants, and metabolite levels were analyzed via untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS). The Chinese version of the Insomnia Severity Index (C-ISI) was employed to evaluate insomnia severity, and the correlations between insomnia severity and these four metabolites were subjected to statistical analysis. Both univariate and multivariate analyses revealed significant metabolic differences between groups. The experimental group showed significantly lower levels of 5-HIAA (FC = 0.947, P = 0.020) and OA (FC = 0.953, P < 0.001), but higher OEA (FC = 1.101, P < 0.001) and EA (FC = 1.026, P < 0.001). C-ISI scores correlated negatively with 5-HIAA (r = -0.380, P = 0.003) and OA (r = -0.448, P < 0.001), and positively with OEA (r = 0.500, P < 0.001) and EA (r = 0.408, P = 0.001). These fecal metabolites associate with insomnia severity and may serve as potential biomarkers for understanding its pathophysiology and developing interventions.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117387"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119341","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

Cost-effective routine pharmaceutical testing using radial flow stream splitting HPLC columns: Quantitative analysis and performance metrics in the analysis of over-the-counter drugs 采用径向流分裂高效液相色谱柱的成本效益常规药物检测：非处方药分析中的定量分析和性能指标。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2026-01-30 DOI: 10.1016/j.jpba.2026.117386

Michalina McDermott , Zachary Sargeant , Christopher E. Karlsen , Feng Li , R. Andrew Shalliker , Jake A. Cravino

The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quantification of active pharmaceutical ingredients, and degradation product analysis. However, traditional HPLC methods are often constrained by pressure limitations at higher flow rates, which can hinder analytical throughput. While recent advancements in column technology have improved performance, they typically exacerbate pressure-related challenges. In this study, we evaluate a novel column technology designed to address these limitations by enabling high-resolution separations at reduced pressures and increased flow rates. Our findings demonstrate that the column, when operated in Radial Flow Stream Splitting (RFS) mode, maintains quantitative accuracy and repeatability while achieving up to a 120 % improvement in separation efficiency and a 30 % reduction in backpressure compared to conventional operation. By way of assaying over-the-counter medication, we have found no difference in the quantitative reliability of the assay when in RFS vs stock mode, despite reducing the analysis time by up to 40 %.

制药行业对快速可靠的分析方法的需求持续增长，超高效液相色谱法（U/HPLC）仍然是杂质分析、活性药物成分定量和降解产物分析的金标准。然而，传统的HPLC方法在高流速下往往受到压力限制，这可能会阻碍分析通量。虽然柱技术的最新进步提高了性能，但它们通常会加剧与压力相关的挑战。在这项研究中，我们评估了一种新的色谱柱技术，该技术旨在解决这些限制，在降低压力和增加流量的情况下实现高分辨率分离。我们的研究结果表明，与传统操作相比，在径向流分裂（RFS）模式下操作的色谱柱，在保持定量准确性和重复性的同时，分离效率提高了120% %，背压降低了30% %。通过分析非处方药物，我们发现在RFS和库存模式下，尽管分析时间减少了40% %，但该分析的定量可靠性没有差异。

{"title":"Cost-effective routine pharmaceutical testing using radial flow stream splitting HPLC columns: Quantitative analysis and performance metrics in the analysis of over-the-counter drugs","authors":"Michalina McDermott , Zachary Sargeant , Christopher E. Karlsen , Feng Li , R. Andrew Shalliker , Jake A. Cravino","doi":"10.1016/j.jpba.2026.117386","DOIUrl":"10.1016/j.jpba.2026.117386","url":null,"abstract":"<div><div>The demand for rapid and reliable analytical methods in the pharmaceutical industry continues to grow, with Ultra-/High-Performance Liquid Chromatography (U/HPLC) remaining the gold standard for impurity profiling, quantification of active pharmaceutical ingredients, and degradation product analysis. However, traditional HPLC methods are often constrained by pressure limitations at higher flow rates, which can hinder analytical throughput. While recent advancements in column technology have improved performance, they typically exacerbate pressure-related challenges. In this study, we evaluate a novel column technology designed to address these limitations by enabling high-resolution separations at reduced pressures and increased flow rates. Our findings demonstrate that the column, when operated in Radial Flow Stream Splitting (RFS) mode, maintains quantitative accuracy and repeatability while achieving up to a 120 % improvement in separation efficiency and a 30 % reduction in backpressure compared to conventional operation. By way of assaying over-the-counter medication, we have found no difference in the quantitative reliability of the assay when in RFS vs stock mode, despite reducing the analysis time by up to 40 %.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117386"},"PeriodicalIF":3.1,"publicationDate":"2026-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119296","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