Journal of separation science最新文献

{"title":"A Metabolism-Oriented Strategy for Systematic Analysis of the Pharmacological Material Basis of Classic Traditional Chinese Medicine Formulas: Taking Wuao Sanzi Decoction as an Example","authors":"Jin-Jin Liao,&nbsp;Yang-Dan Liu,&nbsp;Lan-Ying Gao,&nbsp;Gui-Zhong Xin,&nbsp;Yuan Cao,&nbsp;Zi-Qi Shi","doi":"10.1002/jssc.70521","DOIUrl":"10.1002/jssc.70521","url":null,"abstract":"<div>\u0000 \u0000 <p>Deciphering the pharmacodynamic material basis of classical traditional Chinese medicine (TCM) formulas remains a critical challenge, owing to their multicomponent nature, multitarget interactions, and intricate in vivo metabolic fates. To address this issue, a metabolism-oriented strategy was established and applied to Wuao Sanzi Decoction (WASZD), a classic TCM formula for respiratory diseases such as asthma. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry was used for systematic analysis of the chemicalome, in vivo prototypes, metabolites, and tissue distribution of WASZD. A total of 200 chemical components were identified in vitro, including 102 flavonoids, 29 triterpenoids, 10 steroids, 14 organic acids, and 45 other compounds. Further, 91 prototypes and 212 metabolites in vivo were characterized in biological samples of rats. Key metabolic pathways involved phase I reactions (reduction, oxidation, and demethylation), phase II reactions (glucuronidation, methylation, and sulfation), and gut microbiota-mediated ring cleavage of flavonoids, which generated bioactive small-molecule phenolic acids. Tissue distribution analysis revealed that prototypes and metabolites were predominantly enriched in the ileum, colon, liver, kidney, and lung, with flavonoids and their metabolites identified as the most abundant components, suggesting they are the core bioactive candidates of WASZD. This study not only putatively clarifies the pharmacodynamic material basis of WASZD but also establishes a reproducible technical framework for decoding the complex in vivo behavior of multi-component TCM formulas, facilitating their modernization and rational clinical application.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148865473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Untargeted UHPLC-Q-Exactive Orbitrap MS and Targeted UHPLC-QqQ-MS/MS for Exploring the Effective Components of “Wen Tong Plaster” Cataplasm in Treating Rats With Primary Dysmenorrhea","authors":"Wenjing Chen,&nbsp;Zongtong Yang,&nbsp;Ying Li,&nbsp;Longyun Duan,&nbsp;Tong Zhang,&nbsp;Yufei Li,&nbsp;Zaiyun Sui,&nbsp;Jin Liu,&nbsp;Xinjun Zhang,&nbsp;Beibei Yu,&nbsp;Wenjing Hou","doi":"10.1002/jssc.70508","DOIUrl":"10.1002/jssc.70508","url":null,"abstract":"<div>\u0000 \u0000 <p>“Wen Tong Plaster” is renowned for its therapeutic effects of warming meridians, dispelling cold, and promoting blood circulation. However, its chemical composition and active pharmacological substances have yet to be fully elucidated. In this study, the chemical components of “Wen Tong Plaster” cataplasm and the transdermal components in vivo and in vitro were comprehensively analyzed by mass spectrometry targeted quantification method, and the release kinetics of the main transdermal components in vitro were evaluated, and the concentration-time variation of key active transdermal components in vivo was studied, finally molecular docking technology was used to predict the mechanism of action of key bioactive transdermal components. Using UHPLC-Q-Exactive Orbitrap MS, a total of 129 plant chemical components were found in the “Wen Tong Plaster” cataplasm extract, with 41 components detected as in vitro transdermal compounds. In subcutaneous tissue and uterine tissue, 53 and 36 in vivo transdermal components were characterized, respectively. The release kinetics of “Wen Tong Plaster” cataplasm were evaluated by tetrahydropalmatine, corydaline, and dihydrotanshinone I, three marker components, and fitted to a first-order kinetic model based on integrated release kinetics. After “Wen Tong Plaster” cataplasm administration, tetrahydropalmatine, corydaline, dihydrotanshinone I, and α-cyperone were accurately quantified in the subcutaneous tissue, plasma, and uterus tissue by UHPLC-QqQ-MS/MS, while the concentration-time changes of them in the subcutaneous tissue, plasma, and uterus tissue were investigated. The molecular docking results indicated that all four components exhibited favorable binding affinity with the targets, particularly demonstrating strong binding capacity to the cytochrome P450 1A1 target, suggesting it as a potential therapeutic target for “Wen Tong Plaster” cataplasm. In summary, the study revealed the pharmacodynamic material basis and in vitro release kinetics of “Wen Tong Plaster” cataplasm, providing a theoretical foundation and data support for further research on the mechanism of action and clinical application.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148865555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Wild-Simulated Ginseng Powder, Cultivated Ginseng Powder, Cultivated Ginseng Decoction Pieces, and Baoyuan Decoction Containing Wild-Simulated Ginseng and Cultivated Ginseng by UHPLC-Q-TOF-MS/MS and Non-Targeted Metabolite Profiling.","authors":"Han Li, Yute Zhong, Wenjing Gao, Dan Wu, Weijie Li, Xiaowei Cui, Wei Kang, Haiyu Xu, Ping Wang","doi":"10.1002/jssc.70516","DOIUrl":"10.1002/jssc.70516","url":null,"abstract":"<p><p>Wild-simulated ginseng (linxiashen, LXS) powder, farm-grown ginseng (yuanshen, YS) powder, and farm-grown ginseng decoction pieces (YSDP), as well as the Baoyuan decoction prepared from them (BYD-LXS, BYD-YS, and BYD-YSDP) by boiling, exhibit certain metabolic differences in their chemical composition. To elucidate the holistic molecular profile and differences, a reversed-phase UHPLC-Q-TOF-MS^E metabolomics strategy with an ACQUITY HSS T3 stationary phase was applied to characterize and discover differential chemical markers. In total, 293, 663, 311, and 645 compounds (including 22, 13, 24, and 28 rare ginsenosides) were annotated in LXS/YS, BYD-LXS/BYD-YS, YS/YSDP, and BYD-YS/BYD-YSDP respectively. Based on PCA and OPLS-DA, 87, 222, 95, and 168 chemical markers (including 8, 7, 13, and 3 rare ginsenosides) were annotated in LXS/YS, BYD-LXS/BYD-YS, YS/YSDP, and BYD-YS/BYD-YSDP decoctions, respectively. Secondary metabolites such as triterpene saponins were found to be present at higher levels in LXS decoction and BYD-LXS decoction. The amount of components eluted from YSDP decoction was significantly higher than that from the decoction of YS powder. In addition, Support Vector Machine and Back Propagation Neural Network models with feature selection nested within cross-validation were constructed to classify LXS/YS, BYD-LXS/BYD-YS, YS/YSDP, and BYD-YS/BYD-YSDP decoctions, achieving 100%, 100%, 100%, and 94.44% classification accuracies, respectively. This study provides a basis for the quality assessment of ginseng and Baoyuan decoction, as well as for the development and utilization of rare ginsenosides.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":"e70516"},"PeriodicalIF":3.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13576869/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148969345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Five-Dimensional \"Spider-Web\" Strategy for Quality Markers Identification of Traditional Chinese Medicine Compound Preparations Based on an Integrated Characteristic Network.","authors":"Ji-Yan Li, Fang-Di Hu, Juan Chen","doi":"10.1002/jssc.70528","DOIUrl":"10.1002/jssc.70528","url":null,"abstract":"<p><p>Given the complex composition of traditional Chinese medicine (TCM) formulas, quality evaluation cannot rely on a single component or a limited few. Therefore, a five-dimensional quality marker (Q-marker) identification system encompassing \"effectiveness, specificity, transferability, measurability, and formula compatibility\" was developed and subsequently applied to Fufang Huangqi Jianpi oral liquid (FFHQJPOL) as an example. Using UHPLC-Q-TOF/MS, 62 compounds were identified in the preparation. The measurability dimension was constructed based on 17 major constituents derived from 23 common peaks in the chromatographic fingerprints of 10 batches. Serum pharmacochemistry via UHPLC-Q-TOF/MS revealed 32 prototype components absorbed into the bloodstream, thereby establishing the transferability dimension. Network pharmacology approaches were applied to construct a \"component-target-disease-pathway\" network, defining the effectiveness dimension. A topological network of compounds and herbs was established to depict the distribution pattern of the candidate compounds across various herbs, thereby revealing specificity. The compound compatibility dimension was established based on the function and proportional weight of its active ingredients in the formula. Ultimately, through regression area (RA) and coefficient of variation (CV) calculation, 3-hydroxy-9,10-dimethoxypterocarpan, ononin, zizyphusine, calycosin-7-O-glucoside, and sinapine were initially identified as the key Q-markers. It is worth mentioning that astragaloside IV and astragaloside II were additionally incorporated into the Q-markers system based on UHPLC-Q-TOF/MS analysis and their known pharmacological activities, ultimately establishing seven key Q-markers for FFHQJPOL. This strategy provided a multi‑dimensional integrated paradigm for Q‑markers discovery in TCM compound preparations, facilitating the transition from single‑index to system‑integrated quality control.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":"e70528"},"PeriodicalIF":3.6,"publicationDate":"2026-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13584845/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148990492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bridging the Gap Between Chemiluminescence Immunoassay and Liquid Chromatography-Tandem Mass Spectrometry for Infliximab Monitoring: Method Validation and Clinical Comparability Study","authors":"Ying Xia,&nbsp;Ming-Mei Zhu,&nbsp;Meng-Qing Xiao,&nbsp;Chen-Chun Zhong,&nbsp;Yu-Jun Zhou,&nbsp;Yi-Hao Xue,&nbsp;Shi-Wei Zheng,&nbsp;Li Li,&nbsp;Feng Chen","doi":"10.1002/jssc.70515","DOIUrl":"https://doi.org/10.1002/jssc.70515","url":null,"abstract":"<div>\u0000 \u0000 <p>Infliximab (IFX) is a cornerstone biologic for pediatric inflammatory bowel disease, where therapeutic drug monitoring (TDM) is essential but complicated by inter-platform variability. Systematic comparison of automated immunoassays with reference methods is therefore warranted. This study aimed to validate an in-house chemiluminescence immunoassay (CLIA) for IFX quantification and evaluate its agreement with liquid chromatography-tandem mass spectrometry (LC-MS/MS). A CLIA kit was validated by assessing the limit of blank, specificity, linearity, accuracy, precision, matrix equivalence, and stability. IFX concentrations in 52 pediatric plasma samples were measured by CLIA and a reference LC-MS/MS method. Comparability was evaluated using linear regression, Passing-Bablok regression, and Bland-Altman analysis. CLIA showed excellent performance (lower limit of quantification: 0.640 µg/mL; specificity &lt;10.0% interference; reportable range: 0.640–462 µg/mL). LC-MS/MS yielded higher concentrations than the CLIA (median: 6.25 vs. 2.87 µg/mL). Strong correlation was observed (<i>r</i> = 0.9101) with no significant deviation from linearity (Passing-Bablok, <i>p</i> &gt; 0.05). However, Bland-Altman revealed substantial mean relative bias (67.9%) and poor categorical agreement (52% concordance) within the therapeutic range (3–7 µg/mL), indicating non-interchangeability. We validated a rapid, automated CLIA suitable for high-throughput TDM. Despite strong correlation, significant quantitative differences preclude direct result substitution, underscoring the need for method-specific therapeutic thresholds.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148849359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bridging the Gap Between Chemiluminescence Immunoassay and Liquid Chromatography-Tandem Mass Spectrometry for Infliximab Monitoring: Method Validation and Clinical Comparability Study","authors":"Ying Xia,&nbsp;Ming-Mei Zhu,&nbsp;Meng-Qing Xiao,&nbsp;Chen-Chun Zhong,&nbsp;Yu-Jun Zhou,&nbsp;Yi-Hao Xue,&nbsp;Shi-Wei Zheng,&nbsp;Li Li,&nbsp;Feng Chen","doi":"10.1002/jssc.70515","DOIUrl":"https://doi.org/10.1002/jssc.70515","url":null,"abstract":"<div>\u0000 \u0000 <p>Infliximab (IFX) is a cornerstone biologic for pediatric inflammatory bowel disease, where therapeutic drug monitoring (TDM) is essential but complicated by inter-platform variability. Systematic comparison of automated immunoassays with reference methods is therefore warranted. This study aimed to validate an in-house chemiluminescence immunoassay (CLIA) for IFX quantification and evaluate its agreement with liquid chromatography-tandem mass spectrometry (LC-MS/MS). A CLIA kit was validated by assessing the limit of blank, specificity, linearity, accuracy, precision, matrix equivalence, and stability. IFX concentrations in 52 pediatric plasma samples were measured by CLIA and a reference LC-MS/MS method. Comparability was evaluated using linear regression, Passing-Bablok regression, and Bland-Altman analysis. CLIA showed excellent performance (lower limit of quantification: 0.640 µg/mL; specificity &lt;10.0% interference; reportable range: 0.640–462 µg/mL). LC-MS/MS yielded higher concentrations than the CLIA (median: 6.25 vs. 2.87 µg/mL). Strong correlation was observed (<i>r</i> = 0.9101) with no significant deviation from linearity (Passing-Bablok, <i>p</i> &gt; 0.05). However, Bland-Altman revealed substantial mean relative bias (67.9%) and poor categorical agreement (52% concordance) within the therapeutic range (3–7 µg/mL), indicating non-interchangeability. We validated a rapid, automated CLIA suitable for high-throughput TDM. Despite strong correlation, significant quantitative differences preclude direct result substitution, underscoring the need for method-specific therapeutic thresholds.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148848880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the Active Components of Buqi-Zhitong-Decoction and Its Therapeutic Potential for Chronic Fatigue Syndrome Based on LC-MS/MS and Network Pharmacology","authors":"Yan Liu,&nbsp;Yanli Xin,&nbsp;Xulong Wang,&nbsp;Lixia Wen,&nbsp;Lei Guo,&nbsp;Wenbin He,&nbsp;Qinqing Li","doi":"10.1002/jssc.70510","DOIUrl":"https://doi.org/10.1002/jssc.70510","url":null,"abstract":"<div>\u0000 \u0000 <p>Chronic fatigue syndrome (CFS) is a complex condition that requires effective intervention and treatment. Buqi-Zhitong-Decoction (BQZTD), an empirical formula, has been used clinically to treat CFS and has shown good clinical results. However, the underlying bioactive components and mechanism of action for its treatment of CFS remain unclear. This study is based on the serum pharmacochemistry method, utilizing LC-MS/MS technology for detection and analysis of BQZTD water extracts and their components entering the bloodstream. A total of 98 compounds were preliminarily identified from the BQZTD. Furthermore, 44 ingredients were detected in rat serum, including 19 prototype components and 25 metabolite components. The results of network pharmacology analysis indicated that the BQZTD might improve CFS by regulating immune, inflammatory, neuroendocrine system, and energy metabolism-related pathways. In addition, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) plays a central role in the regulatory network. Molecular docking simulations with six potential active components in serum revealed that all of them exhibit high affinity for PTGS2, with compound Isoformononetin showing the strongest binding affinity. This study represents the first comprehensive identification of the chemical components, targets, and signaling pathways of BQZTD, clarifying its potential mechanism of action in treating CFS. The research findings provide a reference basis for pharmacodynamic studies and quality control.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148849360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the Active Components of Buqi-Zhitong-Decoction and Its Therapeutic Potential for Chronic Fatigue Syndrome Based on LC-MS/MS and Network Pharmacology","authors":"Yan Liu,&nbsp;Yanli Xin,&nbsp;Xulong Wang,&nbsp;Lixia Wen,&nbsp;Lei Guo,&nbsp;Wenbin He,&nbsp;Qinqing Li","doi":"10.1002/jssc.70510","DOIUrl":"https://doi.org/10.1002/jssc.70510","url":null,"abstract":"<div>\u0000 \u0000 <p>Chronic fatigue syndrome (CFS) is a complex condition that requires effective intervention and treatment. Buqi-Zhitong-Decoction (BQZTD), an empirical formula, has been used clinically to treat CFS and has shown good clinical results. However, the underlying bioactive components and mechanism of action for its treatment of CFS remain unclear. This study is based on the serum pharmacochemistry method, utilizing LC-MS/MS technology for detection and analysis of BQZTD water extracts and their components entering the bloodstream. A total of 98 compounds were preliminarily identified from the BQZTD. Furthermore, 44 ingredients were detected in rat serum, including 19 prototype components and 25 metabolite components. The results of network pharmacology analysis indicated that the BQZTD might improve CFS by regulating immune, inflammatory, neuroendocrine system, and energy metabolism-related pathways. In addition, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) plays a central role in the regulatory network. Molecular docking simulations with six potential active components in serum revealed that all of them exhibit high affinity for PTGS2, with compound Isoformononetin showing the strongest binding affinity. This study represents the first comprehensive identification of the chemical components, targets, and signaling pathways of BQZTD, clarifying its potential mechanism of action in treating CFS. The research findings provide a reference basis for pharmacodynamic studies and quality control.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148848881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation and Implementation of Analyte Protectants in the Gas Chromatography–Tandem Mass Spectrometry Analysis of Benzodiazepines and Opioids in Whole Blood Extracts","authors":"Michal P. Dybowski,&nbsp;Krystian J. Siwek","doi":"10.1002/jssc.70517","DOIUrl":"https://doi.org/10.1002/jssc.70517","url":null,"abstract":"<div>\u0000 \u0000 <p>The determination of benzodiazepines and opioids in whole blood by gas chromatography (GC) remains challenging due to low analyte concentrations and matrix-related effects that may compromise analytical response. In this study, the use of analyte protectants (APs) was investigated as a means of enhancing detector response in GC–tandem mass spectrometry (GC–MS/MS) analysis of selected benzodiazepines and opioids following Quick, Easy, Cheap, Effective, Rugged, and Safe-based sample preparation. A series of high-boiling compounds, including linear alcohols, amines, carboxylic acids, and polyethylene glycols (PEGs), were evaluated as APs capable of modulating the chromatographic response. Among the tested compounds, PEG-400 and PEG-600 provided the most consistent signal enhancement, increasing responses of benzodiazepines by 413%–475% and opioids by up to 628% under optimized conditions. Long-chain amines showed the highest enhancement for selected opioids, reaching up to 1080%. The influence of key experimental parameters on APs efficiency was systematically assessed, demonstrating that acetonitrile as the extraction solvent, elevated injector temperatures (up to 325°C), and low initial oven temperatures favored signal enhancement. The developed GC–MS/MS procedure exhibited satisfactory analytical performance, including high linearity (<i>R</i>² &gt; 0.998), good precision (relative standard deviation &lt;5%), and low limits of quantification. Application of the approach to authentic whole-blood samples confirmed that signal enhancement can also be achieved in complex biological matrices, although its magnitude was reduced by the presence of residual matrix constituents. In particular, bromazepam exhibited a 593% increase in signal following the addition of PEG-400. The results demonstrate that the implementation of APs can serve as a simple means of improving analytical response in GC–MS/MS analysis of benzodiazepines and opioids without modification of routine sample preparation procedures. The findings further contribute to the understanding of protectant-mediated signal enhancement mechanisms in the analysis of biological samples.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148823341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation and Implementation of Analyte Protectants in the Gas Chromatography–Tandem Mass Spectrometry Analysis of Benzodiazepines and Opioids in Whole Blood Extracts","authors":"Michal P. Dybowski,&nbsp;Krystian J. Siwek","doi":"10.1002/jssc.70517","DOIUrl":"https://doi.org/10.1002/jssc.70517","url":null,"abstract":"<div>\u0000 \u0000 <p>The determination of benzodiazepines and opioids in whole blood by gas chromatography (GC) remains challenging due to low analyte concentrations and matrix-related effects that may compromise analytical response. In this study, the use of analyte protectants (APs) was investigated as a means of enhancing detector response in GC–tandem mass spectrometry (GC–MS/MS) analysis of selected benzodiazepines and opioids following Quick, Easy, Cheap, Effective, Rugged, and Safe-based sample preparation. A series of high-boiling compounds, including linear alcohols, amines, carboxylic acids, and polyethylene glycols (PEGs), were evaluated as APs capable of modulating the chromatographic response. Among the tested compounds, PEG-400 and PEG-600 provided the most consistent signal enhancement, increasing responses of benzodiazepines by 413%–475% and opioids by up to 628% under optimized conditions. Long-chain amines showed the highest enhancement for selected opioids, reaching up to 1080%. The influence of key experimental parameters on APs efficiency was systematically assessed, demonstrating that acetonitrile as the extraction solvent, elevated injector temperatures (up to 325°C), and low initial oven temperatures favored signal enhancement. The developed GC–MS/MS procedure exhibited satisfactory analytical performance, including high linearity (<i>R</i>² &gt; 0.998), good precision (relative standard deviation &lt;5%), and low limits of quantification. Application of the approach to authentic whole-blood samples confirmed that signal enhancement can also be achieved in complex biological matrices, although its magnitude was reduced by the presence of residual matrix constituents. In particular, bromazepam exhibited a 593% increase in signal following the addition of PEG-400. The results demonstrate that the implementation of APs can serve as a simple means of improving analytical response in GC–MS/MS analysis of benzodiazepines and opioids without modification of routine sample preparation procedures. The findings further contribute to the understanding of protectant-mediated signal enhancement mechanisms in the analysis of biological samples.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"49 9","pages":""},"PeriodicalIF":3.6,"publicationDate":"2026-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148823400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}