Pub Date : 2026-02-11DOI: 10.1016/j.jpba.2026.117412
Yang Han, Jindong Chen, Guangxue Chen, Ligang Shan, Yifan Qiu, Fan Wang, Wenyan Wang
Insomnia is a common sleep issue among pregnant women. This study aimed to explore the metabolic characteristics of pregnant women with insomnia and identify potential biomarkers through plasma and cerebrospinal fluid (CSF) metabolomics analysis. A total of 423 pregnant women participated in the plasma sample collection, and 89 provided CSF samples during delivery. Plasma metabolomics analysis was conducted using UPLC-Q Exactive Orbitrap high-resolution mass spectrometry. Furtherly, targeted amino acid and neurotransmitter analysis in plasma and CSF using validated LC-MS/MS methods. Multivariate statistical methods were employed for data analysis. The results indicated that the incidences of insomnia were 6.74 %, 14.0 %, and 50.0 % in the first, second, and third trimesters, respectively. Plasma metabolomics analysis revealed significant alterations in amino acid metabolism pathways in pregnant women with insomnia. In the following targeted amino acid analysis, alanine demonstrated the highest diagnostic value (AUC: 0.914) in the first trimester, remaining significant in the third trimester. CSF analysis showed elevated glutamine, histidine, methionine, 5-hydroxyindole acetic acid, and dopamine in pregnant women with insomnia. In conclusion, pregnant women with late-pregnancy insomnia exhibited specific amino acid metabolism disorders during the first trimester, indicating potential value of early intervention in preventing pregnancy insomnia. Disrupted amino acid and neurotransmitter metabolism may be key features of pregnancy-related insomnia.
{"title":"Integrating plasma metabolomics using UPLC-Q Exactive Orbitrap HRMS and targeted amino acid and neurotransmitter analysis in plasma and cerebrospinal fluid by LC-MS/MS to explore biomarkers in pregnancy women with insomnia.","authors":"Yang Han, Jindong Chen, Guangxue Chen, Ligang Shan, Yifan Qiu, Fan Wang, Wenyan Wang","doi":"10.1016/j.jpba.2026.117412","DOIUrl":"https://doi.org/10.1016/j.jpba.2026.117412","url":null,"abstract":"<p><p>Insomnia is a common sleep issue among pregnant women. This study aimed to explore the metabolic characteristics of pregnant women with insomnia and identify potential biomarkers through plasma and cerebrospinal fluid (CSF) metabolomics analysis. A total of 423 pregnant women participated in the plasma sample collection, and 89 provided CSF samples during delivery. Plasma metabolomics analysis was conducted using UPLC-Q Exactive Orbitrap high-resolution mass spectrometry. Furtherly, targeted amino acid and neurotransmitter analysis in plasma and CSF using validated LC-MS/MS methods. Multivariate statistical methods were employed for data analysis. The results indicated that the incidences of insomnia were 6.74 %, 14.0 %, and 50.0 % in the first, second, and third trimesters, respectively. Plasma metabolomics analysis revealed significant alterations in amino acid metabolism pathways in pregnant women with insomnia. In the following targeted amino acid analysis, alanine demonstrated the highest diagnostic value (AUC: 0.914) in the first trimester, remaining significant in the third trimester. CSF analysis showed elevated glutamine, histidine, methionine, 5-hydroxyindole acetic acid, and dopamine in pregnant women with insomnia. In conclusion, pregnant women with late-pregnancy insomnia exhibited specific amino acid metabolism disorders during the first trimester, indicating potential value of early intervention in preventing pregnancy insomnia. Disrupted amino acid and neurotransmitter metabolism may be key features of pregnancy-related insomnia.</p>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"117412"},"PeriodicalIF":3.1,"publicationDate":"2026-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146213586","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 : 2026-02-10DOI: 10.1016/j.jpba.2026.117410
Jiyu Xu , Mingyao Wang , Zhifei Hou , Jie Kong , Mengge Zhang , Yizhuo Tian , Jiangfeng Liu , Juntao Yang , Huaping Dai
Chronic inhalation of coal dust causes coal workers’ pneumoconiosis (CWP), which is one of the leading occupational diseases. Coal workers’ pneumoconiosis is currently incurable, posing a serious public health threat. Identifying the underlying molecular mechanisms of CWP is critical to overcome this challenge. Nowadays, metabolomics has bridged underlying molecular alterations with disease progression, providing a useful tool for researching the pathogenesis and finding biomarkers. In this study, a comprehensive view of metabolic characterization of serum from CWP patients at all stages was provided using untargeted metabolomic analysis. As a result, when compared to healthy controls, the specific alteration patterns of each stage were observed. The results showed arginine and cortisol could be core metabolites in CWP progression. Moreover, five metabolites that significantly changed when going from “solely chronic coal-dust exposure” to an early stage were screened out as potential biomarkers. The receiver operating characteristic results were 0.691–0.862 (individual) and 0.884–0.907 (combined). These findings will benefit the application of metabolomics to understand the pathological mechanism and identify diagnostic biomarkers for CWP.
{"title":"Serum metabolomics reveal metabolic changes in coal workers’ pneumoconiosis progression","authors":"Jiyu Xu , Mingyao Wang , Zhifei Hou , Jie Kong , Mengge Zhang , Yizhuo Tian , Jiangfeng Liu , Juntao Yang , Huaping Dai","doi":"10.1016/j.jpba.2026.117410","DOIUrl":"10.1016/j.jpba.2026.117410","url":null,"abstract":"<div><div>Chronic inhalation of coal dust causes coal workers’ pneumoconiosis (CWP), which is one of the leading occupational diseases. Coal workers’ pneumoconiosis is currently incurable, posing a serious public health threat. Identifying the underlying molecular mechanisms of CWP is critical to overcome this challenge. Nowadays, metabolomics has bridged underlying molecular alterations with disease progression, providing a useful tool for researching the pathogenesis and finding biomarkers. In this study, a comprehensive view of metabolic characterization of serum from CWP patients at all stages was provided using untargeted metabolomic analysis. As a result, when compared to healthy controls, the specific alteration patterns of each stage were observed. The results showed arginine and cortisol could be core metabolites in CWP progression. Moreover, five metabolites that significantly changed when going from “solely chronic coal-dust exposure” to an early stage were screened out as potential biomarkers. The receiver operating characteristic results were 0.691–0.862 (individual) and 0.884–0.907 (combined). These findings will benefit the application of metabolomics to understand the pathological mechanism and identify diagnostic biomarkers for CWP.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117410"},"PeriodicalIF":3.1,"publicationDate":"2026-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192204","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 : 2026-02-10DOI: 10.1016/j.jpba.2026.117411
Burçak Demi̇rbakan , Ahmet Çeti̇nkaya , Evrim Güneş Altuntaş , Mehmet Altay Ünal , Mustafa Kemal Sezgi̇ntürk , Sibel A. Özkan
A novel electrochemical biosensor was constructed for the ultrasensitive detection of aflatoxin B1 (AFB1) in food samples. The biosensor design was based on a 3-mercaptopropionic acid (3-MPA)-modified self-assembled monolayer (SAM) constructed on disposable gold screen-printed electrodes (SPE-Au). Covalent attachment of anti-AFB1 antibodies was achieved on the functionalized electrode surface, and the modification process at each stage was analyzed using CV and EIS techniques. 3-MPA concentration was optimized to enhance analytical performance. The biosensor exhibited a wide linear detection range of 0.1–250 pg/mL with a calculated limit of detection (LOD) of 0.94 pg/mL and limit of quantification (LOQ) of 3.14 pg/mL. It also demonstrated high reproducibility and excellent selectivity toward AFB1. Finally, the biosensor was successfully applied to real food samples, including milk, rice, peanuts, and chili pepper, confirming its reliability and potential for practical food safety monitoring.
{"title":"3-mercaptopropionic acid agent-based biosensor system using gold-screen printed electrode for aflatoxin B1 detection","authors":"Burçak Demi̇rbakan , Ahmet Çeti̇nkaya , Evrim Güneş Altuntaş , Mehmet Altay Ünal , Mustafa Kemal Sezgi̇ntürk , Sibel A. Özkan","doi":"10.1016/j.jpba.2026.117411","DOIUrl":"10.1016/j.jpba.2026.117411","url":null,"abstract":"<div><div>A novel electrochemical biosensor was constructed for the ultrasensitive detection of aflatoxin B1 (AFB1) in food samples. The biosensor design was based on a 3-mercaptopropionic acid (3-MPA)-modified self-assembled monolayer (SAM) constructed on disposable gold screen-printed electrodes (SPE-Au). Covalent attachment of anti-AFB1 antibodies was achieved on the functionalized electrode surface, and the modification process at each stage was analyzed using CV and EIS techniques. 3-MPA concentration was optimized to enhance analytical performance. The biosensor exhibited a wide linear detection range of 0.1–250 pg/mL with a calculated limit of detection (LOD) of 0.94 pg/mL and limit of quantification (LOQ) of 3.14 pg/mL. It also demonstrated high reproducibility and excellent selectivity toward AFB1. Finally, the biosensor was successfully applied to real food samples, including milk, rice, peanuts, and chili pepper, confirming its reliability and potential for practical food safety monitoring.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117411"},"PeriodicalIF":3.1,"publicationDate":"2026-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192139","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 : 2026-02-10DOI: 10.1016/j.jpba.2026.117403
Shuyi Lv , Jiayi Lin , Yu Sun , Mingqin Kang , Xin Li , Lili Song , Yubo Li
Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by symmetrical, erosive polyarthritis and multi-organ involvement, with renal complications posing a life-threatening risk. Paeoniae Radix Alba, known as BaiShao (BS) in Chinese, is a widely used herbal medicine for clinical RA treatment, yet its potential in preventing RA-associated renal damage and its underlying active ingredients remain elusive. To address this knowledge gap, we employed a type II collagen-induced arthritis (CIA) rat model. Biochemical assays and histopathological analyses confirmed that BS exerted robust renoprotective effects in CIA rats. Serum and urine metabolomics identified 48 renal damage-related biomarkers, 18 of which showed distinct regulatory trends following BS intervention. Meanwhile, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was used to characterize 34 chemical components in BS. Through an integrative approach that combines two-way orthogonal partial least squares (O2PLS) and Pearson correlation analysis, 18 active ingredients were identified as key mediators of BS-mediated renoprotection. Collectively, this study establishes a novel metabolomics-O2PLS strategy for discovering active ingredients in BS, laying a foundation for understanding the mechanism underlying the prevention of RA-associated renal damage mediated by BS and providing insights for the development of novel preventive therapeutics.
{"title":"Metabolomics technology combined with O2PLS analysis reveals the active components of Paeoniae Radix Alba in preventing renal damage in rheumatoid arthritis","authors":"Shuyi Lv , Jiayi Lin , Yu Sun , Mingqin Kang , Xin Li , Lili Song , Yubo Li","doi":"10.1016/j.jpba.2026.117403","DOIUrl":"10.1016/j.jpba.2026.117403","url":null,"abstract":"<div><div>Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by symmetrical, erosive polyarthritis and multi-organ involvement, with renal complications posing a life-threatening risk. <em>Paeoniae Radix Alba</em>, known as BaiShao (BS) in Chinese, is a widely used herbal medicine for clinical RA treatment, yet its potential in preventing RA-associated renal damage and its underlying active ingredients remain elusive. To address this knowledge gap, we employed a type II collagen-induced arthritis (CIA) rat model. Biochemical assays and histopathological analyses confirmed that BS exerted robust renoprotective effects in CIA rats. Serum and urine metabolomics identified 48 renal damage-related biomarkers, 18 of which showed distinct regulatory trends following BS intervention. Meanwhile, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was used to characterize 34 chemical components in BS. Through an integrative approach that combines two-way orthogonal partial least squares (O2PLS) and Pearson correlation analysis, 18 active ingredients were identified as key mediators of BS-mediated renoprotection. Collectively, this study establishes a novel metabolomics-O2PLS strategy for discovering active ingredients in BS, laying a foundation for understanding the mechanism underlying the prevention of RA-associated renal damage mediated by BS and providing insights for the development of novel preventive therapeutics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117403"},"PeriodicalIF":3.1,"publicationDate":"2026-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192140","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 : 2026-02-09DOI: 10.1016/j.jpba.2026.117409
Fabian E. Bülow , Adam Matkowski , Alexander Weng
Saponins display medicinal properties such as enhancing the drug delivery of biologicals, and boosting immune responses within vaccinations. As plant derived molecules, their phytochemical analysis is key to ensure pharmaceutical quality. The quantification of saponins is typically carried out by high-performance liquid chromatography (HPLC). The aim of this study was the development of a simple thin-layer chromatography (TLC) based method for the quantification of the sapogenin quillaic acid (QA), the triterpenoid backbone of saponins such as QS21 that are used as immunological adjuvants. Following acidic hydrolysis of the saponins, the corresponding sapogenins were derivatized with 4-hydrazino-7-nitro-2,1,3-benzoxadiazole hydrazine (NBD-H) to form fluorescent hydrazones, increasing both the sensitivity and selectivity of the method. The detection of sapogenins with NBD-H is reported here for the first time. The QA-containing TLC bands were identified by mass spectrometry and their quantification was subsequently performed by densitometry. After validation, the method was applied to two plant species from Caryophyllaceae. To verify the plant’s QA contents determined by TLC, a complementary HPLC method was developed. This study presents a new cost-effective method to quantify QA, enabling laboratories with limited resources to monitor plant cultivation and perform phytopharmaceutical quality control.
{"title":"A novel thin layer chromatography based method for the quantification of quillaic acid saponins","authors":"Fabian E. Bülow , Adam Matkowski , Alexander Weng","doi":"10.1016/j.jpba.2026.117409","DOIUrl":"10.1016/j.jpba.2026.117409","url":null,"abstract":"<div><div>Saponins display medicinal properties such as enhancing the drug delivery of biologicals, and boosting immune responses within vaccinations. As plant derived molecules, their phytochemical analysis is key to ensure pharmaceutical quality. The quantification of saponins is typically carried out by high-performance liquid chromatography (HPLC). The aim of this study was the development of a simple thin-layer chromatography (TLC) based method for the quantification of the sapogenin quillaic acid (QA), the triterpenoid backbone of saponins such as QS21 that are used as immunological adjuvants. Following acidic hydrolysis of the saponins, the corresponding sapogenins were derivatized with 4-hydrazino-7-nitro-2,1,3-benzoxadiazole hydrazine (NBD-H) to form fluorescent hydrazones, increasing both the sensitivity and selectivity of the method. The detection of sapogenins with NBD-H is reported here for the first time. The QA-containing TLC bands were identified by mass spectrometry and their quantification was subsequently performed by densitometry. After validation, the method was applied to two plant species from Caryophyllaceae. To verify the plant’s QA contents determined by TLC, a complementary HPLC method was developed. This study presents a new cost-effective method to quantify QA, enabling laboratories with limited resources to monitor plant cultivation and perform phytopharmaceutical quality control.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117409"},"PeriodicalIF":3.1,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192203","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 : 2026-02-09DOI: 10.1016/j.jpba.2026.117408
Yunwang Chen , Long Liu , Yinting Wei , Qing Zhang , Yanping Wang
Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by recurrent episodes of intestinal inflammation and mucosal injury. Qingkui Yuyang decoction (QKY), a clinically validated traditional Chinese medicinal formula, has been widely used in the treatment of UC; however, its pharmacodynamically active constituents and underlying mechanisms of action have not been fully elucidated. In this study, we explored the therapeutic mechanisms of QKY in treating UC by employing a combination of serum pharmacochemistry, network pharmacology, and molecular docking techniques. Initially, using UPLC-Q-Exactive Orbitrap-MS/MS, 28 candidate active compounds in the serum of rats treated with QKY were identified. Subsequently, network pharmacology analysis identified 43 overlapping targets between UC and the active components, and 30 related signaling pathways. Further analysis and molecular docking studies have confirmed that the key active components (Loureirin A, Berberine, Ellagic acid) possess potential for effective therapeutic effects with the core targets (RELA, AKT1). In addition, in vitro experiments demonstrated that QKY significantly downregulated the expression levels of the pro-inflammatory cytokines IL-6 and TNF-α. QKY also markedly reduced the phosphorylation levels of NF-κB p65 and p38 MAPK, as well as the corresponding mRNA expression levels of these signaling molecules. These results suggest that QKY may exert its therapeutic effects on UC by modulating the MAPK and NF-κB signaling pathways, offering a promising strategy for the prevention and treatment of UC.
{"title":"Identification of blood-absorbed components of Qingkui Yuyang decoction and its mechanistic roles in ulcerative colitis based on UPLC-Q-Exactive Orbitrap-MS/MS and network pharmacology","authors":"Yunwang Chen , Long Liu , Yinting Wei , Qing Zhang , Yanping Wang","doi":"10.1016/j.jpba.2026.117408","DOIUrl":"10.1016/j.jpba.2026.117408","url":null,"abstract":"<div><div>Ulcerative colitis (UC) is a chronic inflammatory bowel disease characterized by recurrent episodes of intestinal inflammation and mucosal injury. Qingkui Yuyang decoction (QKY), a clinically validated traditional Chinese medicinal formula, has been widely used in the treatment of UC; however, its pharmacodynamically active constituents and underlying mechanisms of action have not been fully elucidated. In this study, we explored the therapeutic mechanisms of QKY in treating UC by employing a combination of serum pharmacochemistry, network pharmacology, and molecular docking techniques. Initially, using UPLC-Q-Exactive Orbitrap-MS/MS, 28 candidate active compounds in the serum of rats treated with QKY were identified. Subsequently, network pharmacology analysis identified 43 overlapping targets between UC and the active components, and 30 related signaling pathways. Further analysis and molecular docking studies have confirmed that the key active components (Loureirin A, Berberine, Ellagic acid) possess potential for effective therapeutic effects with the core targets (RELA, AKT1). In addition, in vitro experiments demonstrated that QKY significantly downregulated the expression levels of the pro-inflammatory cytokines IL-6 and TNF-α. QKY also markedly reduced the phosphorylation levels of NF-κB p65 and p38 MAPK, as well as the corresponding mRNA expression levels of these signaling molecules. These results suggest that QKY may exert its therapeutic effects on UC by modulating the MAPK and NF-κB signaling pathways, offering a promising strategy for the prevention and treatment of UC.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117408"},"PeriodicalIF":3.1,"publicationDate":"2026-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146180310","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}
Chuanzhi Qingyu (CZQY) recipe is clinically used for the secondary prevention and treatment of cardiovascular diseases associated with atherosclerosis (AS) However, the potential active components and underlying molecular mechanisms require further elucidation. This study established a mouse model of vulnerable AS plaques through a high-fat diet. The efficacy of CZQY in stabilizing these vulnerable plaques was assessed using histological staining techniques. Additionally, biochemical analyses, ELISA assays, small animal ultrasound technology, and immunohistochemistry were employed. Subsequently, the active pharmaceutical ingredients of CZQY were identified through UPLC-Q-TOF-MSE. Finally, a network pharmacology approach, in conjunction with RNA-seq was utilized to predict the targets and mechanisms of CZQY in stabilizing vulnerable AS plaques, which were subsequently validated using Western blot and PCR methods. The research results show that CZQY can significantly improve lipid levels and liver function, reduce serum inflammation and oxidative stress levels, improve vascular function, and stabilize vulnerable AS plaques. The main components of CZQY that stabilize vulnerable AS plaques include flavonoids, terpenoids, and organic acids. The findings from RNA-seq and network pharmacology suggest that CZQY stabilizes vulnerable AS plaques potentially through mechanisms related to lipid metabolism, inflammatory responses, and cell migration, etc. Subsequent RT-qPCR and Western blot analyses confirmed that CZQY significantly modulated the expression of key targets at both mRNA and protein levels. In conclusions, CZQY effectively reduces risk factors associated with vulnerable AS plaques, improves vascular function, and stabilizes these plaques. This may be related to its ability to modulate mechanisms involving lipid metabolism, inflammation, extracellular matrix dynamics, and smooth muscle cell phenotype. These findings provide significant data support for further pharmacological research and clinical applications.
{"title":"Exploring potential mechanism of Chuanzhi Qingyu recipe for vulnerable atherosclerosis plaques based on UPLC-Q-TOF-MS<sup>E</sup>, systems biology strategy, and experimental validation.","authors":"Jiajun Weng, Luyao Huan, Shiyu Zhang, Weiyi Cao, Fengqin Xu, Yanhong Zhang","doi":"10.1016/j.jpba.2026.117404","DOIUrl":"https://doi.org/10.1016/j.jpba.2026.117404","url":null,"abstract":"<p><p>Chuanzhi Qingyu (CZQY) recipe is clinically used for the secondary prevention and treatment of cardiovascular diseases associated with atherosclerosis (AS) However, the potential active components and underlying molecular mechanisms require further elucidation. This study established a mouse model of vulnerable AS plaques through a high-fat diet. The efficacy of CZQY in stabilizing these vulnerable plaques was assessed using histological staining techniques. Additionally, biochemical analyses, ELISA assays, small animal ultrasound technology, and immunohistochemistry were employed. Subsequently, the active pharmaceutical ingredients of CZQY were identified through UPLC-Q-TOF-MSE. Finally, a network pharmacology approach, in conjunction with RNA-seq was utilized to predict the targets and mechanisms of CZQY in stabilizing vulnerable AS plaques, which were subsequently validated using Western blot and PCR methods. The research results show that CZQY can significantly improve lipid levels and liver function, reduce serum inflammation and oxidative stress levels, improve vascular function, and stabilize vulnerable AS plaques. The main components of CZQY that stabilize vulnerable AS plaques include flavonoids, terpenoids, and organic acids. The findings from RNA-seq and network pharmacology suggest that CZQY stabilizes vulnerable AS plaques potentially through mechanisms related to lipid metabolism, inflammatory responses, and cell migration, etc. Subsequent RT-qPCR and Western blot analyses confirmed that CZQY significantly modulated the expression of key targets at both mRNA and protein levels. In conclusions, CZQY effectively reduces risk factors associated with vulnerable AS plaques, improves vascular function, and stabilizes these plaques. This may be related to its ability to modulate mechanisms involving lipid metabolism, inflammation, extracellular matrix dynamics, and smooth muscle cell phenotype. These findings provide significant data support for further pharmacological research and clinical applications.</p>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"274 ","pages":"117404"},"PeriodicalIF":3.1,"publicationDate":"2026-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146776169","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 : 2026-02-07DOI: 10.1016/j.jpba.2026.117405
Yanting Wang , Xiaofei Wang , Ningqi Xia , Hao Chen , Linfeng Zhang , Bin Lu , Diya Lv , Yan Cao
The application of Bio-Layer Interferometry (BLI) is contingent upon the immobilization of highly purified target proteins onto the sensor. The cumbersome and time-consuming nature of traditional protein expression and purification processes restricts the application of BLI in high-throughput screening of traditional Chinese medicine (TCM). This study aims to develop a rapid and efficient BLI-based platform for screening bioactive components in TCM. An integrated platform combining cell-free protein synthesis (CFPS), BLI, and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) was established for efficient TCM bioactive compound discovery. Functional C-X-C chemokine receptor 4 (CXCR4) was synthesized in vitro using a CFPS system, which were then validated by surface plasmon resonance (SPR) and western blotting. Immobilized CXCR4 on NTA biosensors enabled BLI-based high-throughput screening of TCM extracts, followed by target-specific compound recovery and characterized via mass spectrometry. Three bioactive TCM constituents were successfully fished and identified as coptisine, ligustilide, and senkyunolide A. All of them exhibited negligible cytotoxicity at concentrations ranging from 6.25 to 100 μM). Furthermore, ligustilide and senkyunolide A demonstrated certain affinity for CXCR4 with KD of 69.86 μM and 14.7 μM, respectively, and significantly inhibited cell migration. This study is the first identification of ligustilide and senkyunolide A as functional ligands of CXCR4. The established CFPS-BLI-UHPLC-QTOF/MS platform enables efficient discovery of low-toxicity, high-affinity CXCR4-targeting therapeutics from TCM.
{"title":"Construction of bio-layer interferometry biosensors via cell-free synthesized proteins for fishing bioactive compounds from Chinese herbs","authors":"Yanting Wang , Xiaofei Wang , Ningqi Xia , Hao Chen , Linfeng Zhang , Bin Lu , Diya Lv , Yan Cao","doi":"10.1016/j.jpba.2026.117405","DOIUrl":"10.1016/j.jpba.2026.117405","url":null,"abstract":"<div><div>The application of Bio-Layer Interferometry (BLI) is contingent upon the immobilization of highly purified target proteins onto the sensor. The cumbersome and time-consuming nature of traditional protein expression and purification processes restricts the application of BLI in high-throughput screening of traditional Chinese medicine (TCM). This study aims to develop a rapid and efficient BLI-based platform for screening bioactive components in TCM. An integrated platform combining cell-free protein synthesis (CFPS), BLI, and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) was established for efficient TCM bioactive compound discovery. Functional C-X-C chemokine receptor 4 (CXCR4) was synthesized <em>in vitro</em> using a CFPS system, which were then validated by surface plasmon resonance (SPR) and western blotting. Immobilized CXCR4 on NTA biosensors enabled BLI-based high-throughput screening of TCM extracts, followed by target-specific compound recovery and characterized via mass spectrometry. Three bioactive TCM constituents were successfully fished and identified as coptisine, ligustilide, and senkyunolide A. All of them exhibited negligible cytotoxicity at concentrations ranging from 6.25 to 100 μM). Furthermore, ligustilide and senkyunolide A demonstrated certain affinity for CXCR4 with K<sub>D</sub> of 69.86 μM and 14.7 μM, respectively, and significantly inhibited cell migration. This study is the first identification of ligustilide and senkyunolide A as functional ligands of CXCR4. The established CFPS-BLI-UHPLC-QTOF/MS platform enables efficient discovery of low-toxicity, high-affinity CXCR4-targeting therapeutics from TCM.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"273 ","pages":"Article 117405"},"PeriodicalIF":3.1,"publicationDate":"2026-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192141","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 : 2026-02-05DOI: 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.
{"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}
Pub Date : 2026-02-05DOI: 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.
{"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}
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