Journal of separation science最新文献

{"title":"Chemical Derivatization-Based Liquid Chromatography-Mass Spectrometry Method for Fatty Acid Profiling in Biological Samples","authors":"Jingxuan Yang,&nbsp;Yi Wu,&nbsp;Liang Zhao,&nbsp;Meng Li,&nbsp;Wenjun Guo,&nbsp;Yajuan Xu,&nbsp;Yang Wang","doi":"10.1002/jssc.70211","DOIUrl":"https://doi.org/10.1002/jssc.70211","url":null,"abstract":"<div>\u0000 \u0000 <p>Fatty acids (FAs) are important metabolites for various biochemical functions in living organisms, including energy storage, cell structure, and cell signaling. Changes in FAs composition were significantly related to abnormal metabolic levels and body status. In this study, the derivatization-based liquid chromatography-quadrupole-orbitrap mass spectrometry method was established for FA analysis under parallel reaction monitoring mode. The 4-amino-1-benzylpiperidine (4A1BP) was used as a derivatization reagent to amide with FAs. The results showed that protonated molecules of FA+4A1BP–H₂O were observed, and characteristic fragment ions at <i>m</i>/<i>z</i> 174.1280 and 91.0548 were detected in the tandem mass spectrum. The derivatization method was optimized in terms of sample preparation, chromatographic separation, and mass spectrometric conditions. The linearity, stability, and repeatability of the method were also investigated with good results. Finally, we applied the established method to analyze the serum and liver tissue samples, successfully evaluating the efficacy of Qizha Shuangye granules in the treatment of hyperlipidemia rats. The established method has broad utility and great potential in the detection of FAs, which can provide technical support for disease mechanism research, biomarker discovery, as well as food quality assurance, and nutritional value evaluation.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558225","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":"The Quality Control Markers Study of Artificial Tiger Bone Powder Based on Peptidomic by NanoLC-HRMS and UHPLC-QQQ-MS","authors":"Shaowei Hu,&nbsp;Jing Xu,&nbsp;Liying Tang,&nbsp;Fangbo Zhang,&nbsp;Ye Zhao,&nbsp;Jing Li,&nbsp;Hongwei Wu,&nbsp;Rui Zhai,&nbsp;Hongjun Yang","doi":"10.1002/jssc.70208","DOIUrl":"https://doi.org/10.1002/jssc.70208","url":null,"abstract":"<div>\u0000 \u0000 <p>Artificial tiger bone powder (ATBP) is an animal-derived traditional Chinese medicine for treating osteoporosis. The combination of NanoLC-HRMS and UHPLC-QQQ-MS was used to identify and quantify the quality control markers of ATBP. Peptidomics by NanoLC-HRMS was used to characterize the native peptides in ATBP and four individual bone extracts. Five signature peptides from the deer bone extracts were identified in ATBP and validated by comparison with synthetic peptides. These signature peptides were used as quality marker peptides to establish an analytical method using UHPLC-QQQ-MS, which was found to have good specificity, stability, and accuracy. The bioactivity of peptides was determined using in vitro osteoblast proliferation and anti-inflammatory assays. GPQGIAGQRGVV and AFAQLSELH could promote the proliferation and mineralization of MC3T3E1 cells. VDVVGAEALGR and FAVEGPKLVA exhibited anti-inflammatory effects in lipopolysaccharide-treated RAW 264.7 cells. This strategy for screening signature peptides in ATBP exhibits significant translational potential and can be systematically applied to the identification and characterization of quality marker peptides in various animal-derived pharmaceuticals.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144558231","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":"Eco-Friendly Miniaturized Solid Phase Microextraction for Pesticide Analysis in Grapes","authors":"Mereke Alimzhanova,&nbsp;Yerkanat Syrgabek,&nbsp;Pedro Antonio Garcia Encina","doi":"10.1002/jssc.70204","DOIUrl":"https://doi.org/10.1002/jssc.70204","url":null,"abstract":"<div>\u0000 \u0000 <p>A simple and environmentally friendly method based on miniaturized headspace solid-phase microextraction (Mini-HS-SPME) coupled with gas chromatography–mass spectrometry (GC-MS) was developed for the identification and quantification of ten pesticides (atrazine, simazine, prometryn, metribuzin, cyanazine, fluroxypyr-meptyl, tebuconazole, epoxiconazole, quizalofop-methyl, and boscalid) in grape samples. Key extraction parameters were optimized, including extraction temperature, extraction time, selection of the optimal fiber, incubation time, effect of salt, volume of sample, and pH. Final extraction conditions included the use of 200 µL of sample using PDMS 100 µm fiber in headspace mode at 90°C for 30 min. The Mini-HS-SPME coupled with the GC-MS method has been successfully validated in terms of linearity, recovery, and accuracy. The studied linear range of pesticides was 5–1000 ng mL⁻¹. The limit of detection and limit of quantification ranged between 0.09–3.0 ng mL⁻¹ and 0.2–10.1 ng mL⁻¹, respectively. In conclusion, this research presents a method for detecting pesticides in terms of miniaturization, and evaluation of environmental friendliness and applicability using modern GAPI, AGREEprep, and BAGI tools.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144519723","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":"Eco-Friendly Miniaturized Solid Phase Microextraction for Pesticide Analysis in Grapes","authors":"Mereke Alimzhanova,&nbsp;Yerkanat Syrgabek,&nbsp;Pedro Antonio Garcia Encina","doi":"10.1002/jssc.70204","DOIUrl":"https://doi.org/10.1002/jssc.70204","url":null,"abstract":"<div>\u0000 \u0000 <p>A simple and environmentally friendly method based on miniaturized headspace solid-phase microextraction (Mini-HS-SPME) coupled with gas chromatography–mass spectrometry (GC-MS) was developed for the identification and quantification of ten pesticides (atrazine, simazine, prometryn, metribuzin, cyanazine, fluroxypyr-meptyl, tebuconazole, epoxiconazole, quizalofop-methyl, and boscalid) in grape samples. Key extraction parameters were optimized, including extraction temperature, extraction time, selection of the optimal fiber, incubation time, effect of salt, volume of sample, and pH. Final extraction conditions included the use of 200 µL of sample using PDMS 100 µm fiber in headspace mode at 90°C for 30 min. The Mini-HS-SPME coupled with the GC-MS method has been successfully validated in terms of linearity, recovery, and accuracy. The studied linear range of pesticides was 5–1000 ng mL⁻¹. The limit of detection and limit of quantification ranged between 0.09–3.0 ng mL⁻¹ and 0.2–10.1 ng mL⁻¹, respectively. In conclusion, this research presents a method for detecting pesticides in terms of miniaturization, and evaluation of environmental friendliness and applicability using modern GAPI, AGREEprep, and BAGI tools.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144520230","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":"Simultaneous Determination of Seven Benzotriazole UV Stabilizers in Surface Waters by Using Solid-Phase Extraction Coupled With Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry With Electrospray Ionization","authors":"Chen Huifan,&nbsp;Zhou Huimin,&nbsp;Wang Zhenguo,&nbsp;Hu Xialin,&nbsp;Yin Daqiang","doi":"10.1002/jssc.70200","DOIUrl":"https://doi.org/10.1002/jssc.70200","url":null,"abstract":"<div>\u0000 \u0000 <p>Benzotriazole ultraviolet stabilizers (BUVSs) are emerging persistent pollutants, due to the weak polarity and wide log <i>K</i>_ow values, there is no general optimal analytical method for BUVSs. In this study, a solid-phase extraction-ultra performance liquid chromatography tandem mass spectrometry (SPE-UHPLC/MS/MS) method was developed for the simultaneous determination of seven BUVSs in the aqueous environment, based on the generalized electrospray ionization (ESI+) mode. The conditions of ESI-UHPLC-MS/MS detection and SPE were systematically optimized. A 50 mm UHPLC column at a column temperature of 40°C gave the highest response for the target BUVSs using methanol–0.1% (v/v) formic acid solution as the mobile phase. The effects of the four packed SPE cartridges (HLB, HC-C₁₈, LC-C₁₈ and phenyl columns) and sample pH (3.0–9.0) on the SPE enrichment were comparatively analyzed, and the optimum condition was HLB cartridges at sample pH 3.0. The SPE elution solvent was optimized as 5 mL methanol + 5 mL dichloromethane (V:V = 1:1). The seven target BUVSs were quantified by isotope internal standard method, and the linear range of the standard curve was wide (0.1–50 ng∙L⁻¹) with the correlation coefficient <i>R</i>² &gt; 0.998. The recoveries of the spiked standards for the purified water as well as the sample of Huangpu River were 80.6%–105.2% and 53.9%–103.7% (except UV-P: 141.6%–148.7%), respectively. The relative standard deviations (RSDs) of the intra-day were 4.1%–18.0%; and the RSDs of the inter-day were 3.4%–18.8%, respectively. The limits of detection of the method were in the range of 0.050–0.406 ng∙L⁻¹. Compared with the reported studies, this method is more sensitive and stable, and the UHPLC-MS/MS instrument based on the ESI ionization source is more generalizable.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144520226","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":"Simultaneous Determination of Seven Benzotriazole UV Stabilizers in Surface Waters by Using Solid-Phase Extraction Coupled With Ultra-High-Performance Liquid Chromatography Tandem Mass Spectrometry With Electrospray Ionization","authors":"Chen Huifan,&nbsp;Zhou Huimin,&nbsp;Wang Zhenguo,&nbsp;Hu Xialin,&nbsp;Yin Daqiang","doi":"10.1002/jssc.70200","DOIUrl":"https://doi.org/10.1002/jssc.70200","url":null,"abstract":"<div>\u0000 \u0000 <p>Benzotriazole ultraviolet stabilizers (BUVSs) are emerging persistent pollutants, due to the weak polarity and wide log <i>K</i>_ow values, there is no general optimal analytical method for BUVSs. In this study, a solid-phase extraction-ultra performance liquid chromatography tandem mass spectrometry (SPE-UHPLC/MS/MS) method was developed for the simultaneous determination of seven BUVSs in the aqueous environment, based on the generalized electrospray ionization (ESI+) mode. The conditions of ESI-UHPLC-MS/MS detection and SPE were systematically optimized. A 50 mm UHPLC column at a column temperature of 40°C gave the highest response for the target BUVSs using methanol–0.1% (v/v) formic acid solution as the mobile phase. The effects of the four packed SPE cartridges (HLB, HC-C₁₈, LC-C₁₈ and phenyl columns) and sample pH (3.0–9.0) on the SPE enrichment were comparatively analyzed, and the optimum condition was HLB cartridges at sample pH 3.0. The SPE elution solvent was optimized as 5 mL methanol + 5 mL dichloromethane (V:V = 1:1). The seven target BUVSs were quantified by isotope internal standard method, and the linear range of the standard curve was wide (0.1–50 ng∙L⁻¹) with the correlation coefficient <i>R</i>² &gt; 0.998. The recoveries of the spiked standards for the purified water as well as the sample of Huangpu River were 80.6%–105.2% and 53.9%–103.7% (except UV-P: 141.6%–148.7%), respectively. The relative standard deviations (RSDs) of the intra-day were 4.1%–18.0%; and the RSDs of the inter-day were 3.4%–18.8%, respectively. The limits of detection of the method were in the range of 0.050–0.406 ng∙L⁻¹. Compared with the reported studies, this method is more sensitive and stable, and the UHPLC-MS/MS instrument based on the ESI ionization source is more generalizable.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144520225","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":"Study on Quality Markers of Chebulae Fructus Immaturus by the Integration of Network Pharmacology, UHPLC Fingerprint, and Serum Pharmacochemistry","authors":"Qian-Qian Li,&nbsp;Ru Yang,&nbsp;Xing-Ping Luo,&nbsp;Juan Chen","doi":"10.1002/jssc.70199","DOIUrl":"https://doi.org/10.1002/jssc.70199","url":null,"abstract":"<div>\u0000 \u0000 <p>The chemical composition of <i>Chebulae Fructus Immaturus</i>, a traditional Chinese medicine, is complex. Its efficacy relies on the interaction of its components, making quality evaluation based on a single chemical insufficient. This study proposes a multi-dimensional feature network to enhance <i>Chebulae Fructus Immaturus</i> quality assessment, focusing on effectiveness, content, traceability, and specificity. The network pharmacological analysis is employed to elucidate the underlying mechanism by which chemical components in <i>Chebulae Fructus Immaturus</i> exert their therapeutic effects, specifically in terms of heat-clearing, body fluid promotion, and detoxification. UHPLC fingerprinting analysis can systematically parse the overall chemical profile of <i>Chebulae Fructus Immaturus</i>, and in combination with chemometric analysis, based on the screening criterion of variable importance projection value &gt; 1, nine potential differential compounds in <i>Chebulae Fructus Immaturus</i> were screened out from the chemometric analysis. Subsequently, the UHPLC-QTOF-MS method, which features high sensitivity, high selectivity, high precision and is capable of multi-stage mass spectrometry, was employed to qualitatively identify the aforementioned potential differential compounds to accurately reveal the chemical constituents of <i>Chebulae Fructus Immaturus</i>. The rat serum was subjected to pharmacochemical analysis, which revealed the presence of nine prototype components that were absorbed into the bloodstream. The candidate components were identified and the scores for four dimensions were computed, leading to the construction of a multi-dimensional feature network based on the “spider web” model. Key quality markers in <i>Chebulae Fructus Immaturus</i> were identified as ellagic acid, chebulic acid, methyl gallate, and gallic acid. In conjunction with our prior research on the quality markers of <i>Chebulae Fructus</i>. The binding ability of quality markers in <i>Chebulae Fructus Immaturus</i> and <i>Chebulae Fructus</i> to their corresponding core targets was verified using molecular docking. This multi-dimensional approach founded on advanced analytical methods is capable of accurately identifying and validating key <i>Chebulae Fructus Immaturus</i> quality markers.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514833","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":"Development of a Solid Phase Extraction Cartridge-Based Analytical Method for Determination of Free Drug in Nanoliposomal Oncology Drug Formulations","authors":"Wei Zhang,&nbsp;Mark Paciolla,&nbsp;Aastha Chadha,&nbsp;Kaylee Worrell","doi":"10.1002/jssc.70152","DOIUrl":"https://doi.org/10.1002/jssc.70152","url":null,"abstract":"<p>A reliable SPE-free drug testing method was developed for active loading nanoliposome formulations through a systematic development approach. Ultrafiltration (UF), nanoparticle exclusion chromatography (HPLC-nPEC), solid phase extraction (SPE), and size exclusion chromatography (SEC) were screened for liposome-free drug separation. Full-scale method development was then conducted on the selected SPE technique. HLB 3cc (60 mg) SPE cartridges were used for developing a liposome-free drug testing procedure. SPE retention capacity for the Mirati drug was determined (1028 µg per cartridge). SPE testing conditions were studied to achieve a good separation between drug-loaded liposomes and free drug, which include cartridge conditioning, buffer wash steps for liposomal drug elution, organic media wash steps for free drug elution, and sample size. Qualification of this newly developed SPE method has demonstrated sufficient specificity/selectivity, excellent detection linearity (correlation coefficient (<i>R</i>) &gt; 0.999) over a study concentration range (1.0 to 46.9 µg/mL), acceptable LOQ (0.52 µg/mL equivalent to 1.7% of free drug in a liposome formulation at 2.5 mg/mL), good accuracy/recovery (within 90% to 94%) for free drug in spiked samples at 4.5%, 9%, and 18% levels, satisfactory method precision (RSD (<i>n</i> = 6) of &lt;2.0% for free drug analysis), and 2-day solution stability for standard and sample solutions (2–8°C). In comparative sample testing, SPE free drug results were in good agreement with SEC results for testing active loading formulations. A significant difference in free drug detection was observed among SPE, SEC, and HPLC-nPEC methods for testing passive loading formulations.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514764","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":"Development of a Solid Phase Extraction Cartridge-Based Analytical Method for Determination of Free Drug in Nanoliposomal Oncology Drug Formulations","authors":"Wei Zhang,&nbsp;Mark Paciolla,&nbsp;Aastha Chadha,&nbsp;Kaylee Worrell","doi":"10.1002/jssc.70152","DOIUrl":"https://doi.org/10.1002/jssc.70152","url":null,"abstract":"<p>A reliable SPE-free drug testing method was developed for active loading nanoliposome formulations through a systematic development approach. Ultrafiltration (UF), nanoparticle exclusion chromatography (HPLC-nPEC), solid phase extraction (SPE), and size exclusion chromatography (SEC) were screened for liposome-free drug separation. Full-scale method development was then conducted on the selected SPE technique. HLB 3cc (60 mg) SPE cartridges were used for developing a liposome-free drug testing procedure. SPE retention capacity for the Mirati drug was determined (1028 µg per cartridge). SPE testing conditions were studied to achieve a good separation between drug-loaded liposomes and free drug, which include cartridge conditioning, buffer wash steps for liposomal drug elution, organic media wash steps for free drug elution, and sample size. Qualification of this newly developed SPE method has demonstrated sufficient specificity/selectivity, excellent detection linearity (correlation coefficient (<i>R</i>) &gt; 0.999) over a study concentration range (1.0 to 46.9 µg/mL), acceptable LOQ (0.52 µg/mL equivalent to 1.7% of free drug in a liposome formulation at 2.5 mg/mL), good accuracy/recovery (within 90% to 94%) for free drug in spiked samples at 4.5%, 9%, and 18% levels, satisfactory method precision (RSD (<i>n</i> = 6) of &lt;2.0% for free drug analysis), and 2-day solution stability for standard and sample solutions (2–8°C). In comparative sample testing, SPE free drug results were in good agreement with SEC results for testing active loading formulations. A significant difference in free drug detection was observed among SPE, SEC, and HPLC-nPEC methods for testing passive loading formulations.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514837","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":"Study on Quality Markers of Chebulae Fructus Immaturus by the Integration of Network Pharmacology, UHPLC Fingerprint, and Serum Pharmacochemistry","authors":"Qian-Qian Li,&nbsp;Ru Yang,&nbsp;Xing-Ping Luo,&nbsp;Juan Chen","doi":"10.1002/jssc.70199","DOIUrl":"https://doi.org/10.1002/jssc.70199","url":null,"abstract":"<div>\u0000 \u0000 <p>The chemical composition of <i>Chebulae Fructus Immaturus</i>, a traditional Chinese medicine, is complex. Its efficacy relies on the interaction of its components, making quality evaluation based on a single chemical insufficient. This study proposes a multi-dimensional feature network to enhance <i>Chebulae Fructus Immaturus</i> quality assessment, focusing on effectiveness, content, traceability, and specificity. The network pharmacological analysis is employed to elucidate the underlying mechanism by which chemical components in <i>Chebulae Fructus Immaturus</i> exert their therapeutic effects, specifically in terms of heat-clearing, body fluid promotion, and detoxification. UHPLC fingerprinting analysis can systematically parse the overall chemical profile of <i>Chebulae Fructus Immaturus</i>, and in combination with chemometric analysis, based on the screening criterion of variable importance projection value &gt; 1, nine potential differential compounds in <i>Chebulae Fructus Immaturus</i> were screened out from the chemometric analysis. Subsequently, the UHPLC-QTOF-MS method, which features high sensitivity, high selectivity, high precision and is capable of multi-stage mass spectrometry, was employed to qualitatively identify the aforementioned potential differential compounds to accurately reveal the chemical constituents of <i>Chebulae Fructus Immaturus</i>. The rat serum was subjected to pharmacochemical analysis, which revealed the presence of nine prototype components that were absorbed into the bloodstream. The candidate components were identified and the scores for four dimensions were computed, leading to the construction of a multi-dimensional feature network based on the “spider web” model. Key quality markers in <i>Chebulae Fructus Immaturus</i> were identified as ellagic acid, chebulic acid, methyl gallate, and gallic acid. In conjunction with our prior research on the quality markers of <i>Chebulae Fructus</i>. The binding ability of quality markers in <i>Chebulae Fructus Immaturus</i> and <i>Chebulae Fructus</i> to their corresponding core targets was verified using molecular docking. This multi-dimensional approach founded on advanced analytical methods is capable of accurately identifying and validating key <i>Chebulae Fructus Immaturus</i> quality markers.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 7","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144514955","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}