Journal of Chromatography B最新文献

{"title":"Pentafluorobenzyl bromide – A versatile derivatization agent in chromatography and mass spectrometry: II. Analysis of organic acids and bases, and comparison with other perfluorinated reagents","authors":"Dimitrios Tsikas","doi":"10.1016/j.jchromb.2025.124578","DOIUrl":"10.1016/j.jchromb.2025.124578","url":null,"abstract":"<div><div>Analytical derivatization is an important for the vast majority of substances an indispensable sample preparation step for their quantitative GC–MS and GC–MS/MS analysis in biological samples. Pentafluorobenzyl bromide (PFB-Br), pentafluorobenzoyl chloride (PFB-COCl), pentafluorobenzyl hydroxylamine (PFB-NHNH₂), pentafluorophenyl hydrazine (PFPh-ONH₂), pentafluoropropionic anhydride (PFPA), and heptafluorobutyric anhydride (HFBA) are versatile derivatization reagents in analytical chemistry. In the present work, the utility of the above mentioned derivatization reagents for the GC–MS analysis of carboxylic, aldehydic, hydroxylic and amine groups containing analytes including amino acids is reviewed and discussed. Derivatization requires different conditions for solvents, reaction temperature and time, and possibly for catalysts. The perfluorinated derivatives are electrically neutral and best soluble in water-immiscible organic solvents such as toluene. Under negative-ion chemical ionization (NICI) conditions, the perfluorinated derivatives readily and abundantly ionize that allows for sensitive analysis. In addition, the perfluorinated analyte derivatives emerge earlier from GC columns than protiated, thus enabling shorter analysis times. Externally added ²H-, ¹³C-, ¹⁵N and ¹⁸O-isotopologs for use as internal standards undergo similar changes during derivatization, extraction by organic solvents, ionization in the ion-source of GC–MS apparatus and have almost identical retention times with the analytes. Due to selective analytical derivatization, almost all classes of endogenous and exogenous low-molecular-mass analytes, including drugs and inorganic anions such as nitrite, nitrate, carbonate, and (pseudo)halogenides, become accessible to quantitative GC–MS and GC–MS/MS analysis. Thanks the high sensitivity of quantitative analytical methods based on GC–MS and GC–MS/MS, very low amounts of perfluorinated derivatization reagents are consumed. In consideration of the enormously high global warming potential (GWP) of F-containing derivatization reagents, this article discussed a potential abandonment of the use of perfluorinated reagents and their replacement by F-free reagents in GC–MS and GC–MS/MS.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124578"},"PeriodicalIF":2.8,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734726","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel ultra-sensitive LC-MS/MS method for determination of elobixibat in human plasma; Application to a bioequivalence study on healthy volunteers","authors":"Mamdouh R. Rezk ,&nbsp;Michael Soliman ,&nbsp;Huda Shawky ,&nbsp;Kamal A. Badr ,&nbsp;Mina Wadie","doi":"10.1016/j.jchromb.2025.124576","DOIUrl":"10.1016/j.jchromb.2025.124576","url":null,"abstract":"<div><div>Towards a new era for alleviating chronic idiopathic constipation, elobixibat has been recently approved in Japan and completed phase III in clinical trials as a highly potent inhibitor for ileal bile acid transporter. This work provides the field of biomedical analysis and clinical studies with the first bioanalytical method for elobixibat quantitation in real human plasma. A new ultra-sensitive liquid chromatography-tandem mass spectrometric method was developed using elobixibat-d5 as an internal standard. First of all, several preliminary efforts were exerted and directed towards optimizing sample preparation procedures to extract the desired drug at a very low concentration level and to avoid any matrix interference or masking. The trials settled on adopting liquid-liquid extraction using methyl tertiary butyl ether after adding 200 μL of 10 % formic acid to 500 μL plasma sample. Chromatographic separation was then conducted using Kinetex® EVO C₁₈ column with mobile phase composed of acetonitrile and 20 mM ammonium format acidified with 0.1 % formic acid in ratio of 80:20 (<em>v</em>/v) and pumped at 0.6 mL/min. Positive electrospray ionization was adopted for mass acquisition, operated in multiple reactions monitoring (MRM) mode at <em>m</em>/<em>z</em> 696 → 593.1 for elobixibat and m/z 701 → 598.1 for the IS. Full bioanalytical validation as per FDA guidance was done over a range of 20.0–1500.0 pg/mL. The proposed method was successfully exploited for determination of elobixibat in human plasma samples and extended to estimate the pharmacokinetic parameters after administration of a single oral dose of 5 mg elobixibat tablet to thirty healthy volunteers.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124576"},"PeriodicalIF":2.8,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734736","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}

{"title":"Comparative pharmacokinetics of seven propargyl-linked antifolate antibiotics in the mouse","authors":"John Hoody ,&nbsp;Jeremy B. Alverson ,&nbsp;Santosh Keshipeddy ,&nbsp;Patrick A. Barney ,&nbsp;Larissa Walker ,&nbsp;Nathan D. Gibson ,&nbsp;Grant J. Sormunen ,&nbsp;Stephen C. Bergmeier ,&nbsp;Amy C. Anderson ,&nbsp;Dennis L. Wright ,&nbsp;Nigel D. Priestley","doi":"10.1016/j.jchromb.2025.124575","DOIUrl":"10.1016/j.jchromb.2025.124575","url":null,"abstract":"<div><div>Antimicrobial resistance (AMR) to existing antibiotics poses a critical global health challenge, with significant morbidity and mortality from bacterial infections. Methicillin-resistant <em>Staphylococcus aureus</em> (MRSA) and vancomycin-resistant strains (VISA, VRSA) are among the most pressing threats, particularly for vulnerable populations. To combat this crisis, the development of novel therapeutic strategies is imperative.</div><div>We report the pharmacokinetic evaluation of a promising class of propargyl-linked diaminopyrimidine dihydrofolate reductase (DHFR) inhibitors with potent activity against drug-resistant bacteria, including MRSA and VISA strains. Previous studies have demonstrated minimum inhibitory concentration (MIC) values below 1 μg.mL⁻¹ for several compounds in this series. Here, we detail the development and validation of an LC-QQQ bioanalytical method for seven propargyl-linked diaminopyrimidine analogues. Pharmacokinetic studies in a murine model across intravenous (IV), intraperitoneal (IP), and oral (PO) routes revealed substantial variability in parameters such as half-life (t₁_/₂), area under the curve (AUC), and peak plasma concentration (Cmax).</div><div>Compound <strong>38C1</strong> demonstrated favorable solubility, a higher maximum tolerated dose, and oral bioavailability of 20 %, making it a lead candidate. Pharmacokinetic-to-MIC ratio analyses showed that <strong>38C1</strong> maintained plasma concentrations significantly above MIC values for multiple <em>S. aureus</em> strains, including MRSA and VISA.</div><div>These findings highlight <strong>38C1</strong> as a promising antifolate candidate for further development. Ongoing studies will assess its efficacy in infection models and refine delivery strategies to maximize therapeutic potential while mitigating resistance development.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124575"},"PeriodicalIF":2.8,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143799928","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}

{"title":"A novel quality-by-design assisted HPLC-DAD method for the simultaneous quantification of tryptophan, tryptophol, and voriconazole for early diagnosis and prognosis of fungal infections decoding quorum sensing phenomenon","authors":"Ahmed M. Saleh ,&nbsp;Ola A. Saleh ,&nbsp;Rabeay Y.A. Hassan ,&nbsp;Amr M. Badawey ,&nbsp;Hoda M. Marzouk","doi":"10.1016/j.jchromb.2025.124571","DOIUrl":"10.1016/j.jchromb.2025.124571","url":null,"abstract":"<div><div>For the first time, a comprehensive analytical approach is introduced that simultaneously quantifies a metabolic precursor (tryptophan), a quorum-sensing biomarker for fungal infections (tryptophol), and an antifungal drug (voriconazole) within a single platform using reversed-phase high-performance liquid chromatography coupled with diode array detection (HPLC-DAD). The method utilizes a Pursuit PFP column featuring a unique pentafluorinated structure, with a mobile phase of methanol: water (60:40, v/v), a flow rate of 1.0 mL/min, and a detection wavelength set at 254.0 nm. An Analytical Quality-by-Design (AQbD) methodology was employed, incorporating a full factorial design for optimal method development. Validation was performed in accordance with ICH guidelines, demonstrating exceptional linearity (2.0–60.0 μg/mL) for all target analytes, along with high precision, accuracy, and system suitability. Furthermore, the method proved robust and versatile when applied to complex matrices, including spiked human serum and pharmaceutical tablet formulations. Noteworthy is the integration of green and white chemistry principles for evaluating the method's sustainability, representing a significant advancement in analytical technique development. Assessment of greenness, blueness, and whiteness with AGREE, ComplexGAPI index, BAGI and RGB 12 Tools, respectively. This innovative analytical platform provides a powerful tool for the early detection and real-time therapeutic monitoring of fungal infections. By enabling the simultaneous analysis of a metabolic marker, a quorum-sensing specific biomarker, and an antifungal agent, the method advances personalized medicine. It offers a novel, efficient, and sustainable solution for the personalized management of fungal infections, enhancing both diagnostic and therapeutic strategies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124571"},"PeriodicalIF":2.8,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143768008","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}

{"title":"Liquid chromatography in determination of pharmacokinetic properties of compounds in drug discovery process","authors":"Mihajlo V. Jakanovski ,&nbsp;Marko D. Jović ,&nbsp;Mirjana D. Mosić ,&nbsp;Dušanka M. Milojković-Opsenica ,&nbsp;Sandra B. Šegan","doi":"10.1016/j.jchromb.2025.124574","DOIUrl":"10.1016/j.jchromb.2025.124574","url":null,"abstract":"<div><div>Liquid chromatography plays a pivotal role in the determination of pharmacokinetic properties during drug discovery, particularly through the evaluation of lipophilicity. This parameter is essential in drug development, significantly influencing pharmacokinetic and pharmacodynamic behavior of potential drugs. It affects membrane permeability, solubility, distribution, and interaction with biological targets, making it a central focus in the early stages of drug design. Poor lipophilicity-related characteristics are often associated with drug failures, inefficacy, toxicity, and increased development costs. Experimental and computational methods, such as chromatographic techniques and theoretical calculations, are vital for accurately determining lipophilicity. These approaches enable the simulation of biological processes, providing insights into how lipophilicity impacts ADME (absorption, distribution, metabolism, and excretion) properties and supporting the optimization of drug candidates. <em>In silico</em> tools further enhance the efficiency of ADME evaluations, reducing the risk of pharmacokinetic-related failures and streamlining the drug discovery process.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124574"},"PeriodicalIF":2.8,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734787","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}

{"title":"Determination of disulfiram and diethyldithiocarbamate in plasma by high performance liquid chromatography with an electrochemical detection using a diamond electrode","authors":"Masahiro Komeno ,&nbsp;Arisa Ohta ,&nbsp;Naoaki Tanabe ,&nbsp;Hiroko Abe ,&nbsp;Yuya Terashima ,&nbsp;Akiyoshi Saitoh ,&nbsp;Kyohei Higashi","doi":"10.1016/j.jchromb.2025.124568","DOIUrl":"10.1016/j.jchromb.2025.124568","url":null,"abstract":"<div><div>Disulfiram (DSF), a drug approved for the treatment of alcoholism, has been increasing attention as an anti-inflammatory agent via inhibit cytoplasmic FROUNT, a common regulator of chemokine receptor CCR2 and CCR5 signaling. Although the determination of DSF in biological fluids, particularly plasma, is important for evaluating drug efficacy, rapid degradation of DSF by albumin often interferes with its accuracy and reproducibility. In this study, cost-effective, selective, and reproducible high-performance liquid chromatography (HPLC) using an electrochemical detector (ECD) equipped with a diamond electrode was used to quantify DSF and its metabolite, diethyldithiocarbamate (DDTC). The limit of quantification (LOQ) for DSF in an albumin-free solution using the HPLC-ECD method was 0.04 μg/mL; however, DSF was not detected in the serum spiked with DSF. Two forms of DSF metabolites, the free and albumin bound forms of DDTC, were detected as methyl-DDTC (MeDDTC) by methyl iodide treatment. The LOQ of MeDDTC in acetonitrile and serum determined using the HPLC-ECD method were 0.41 μg/mL and 1.22 μg/mL, respectively. Plasma MeDDTC was detected in rats that were orally administered DSF (1000 mg/kg) up to 48 h after treatment. This result suggests that HPLC-ECD with a diamond electrode is useful for determining MeDDTC for DSF monitoring in plasma.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124568"},"PeriodicalIF":2.8,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143698138","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}

{"title":"Comparison of three extraction methods, DMU-SPME, SAFE, and SDE, for the analysis of flavor compounds in soy sauce","authors":"Yun-Jiao Ma ,&nbsp;An-Qi Bi ,&nbsp;Ping Li ,&nbsp;Bei-Wei Zhu ,&nbsp;Ming Du ,&nbsp;Xian-Bing Xu","doi":"10.1016/j.jchromb.2025.124562","DOIUrl":"10.1016/j.jchromb.2025.124562","url":null,"abstract":"<div><div>The dual mode unity solid-phase microextraction (DMU-SPME) was evaluated against traditional solvent-based methods, including solvent-assisted flavor evaporation (SAFE) and simultaneous distillation extraction (SDE), to explore its potential as an alternative. Compared to SAFE and SDE, DMU-SPME is a green extraction technology for volatile organic compounds. The results demonstrated that DMU-SPME significantly outperformed both SAFE and SDE in extracting medium-volatility compounds (retention time of 20–40 min), identifying 55 unique volatiles. With the exception of hydrocarbons, DMU-SPME exhibited superior extraction efficiency and significantly better stability compared to both SAFE and SDE methods. Thus, DMU-SPME proves to be a promising alternative for the comprehensive extraction of volatile organic compounds.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124562"},"PeriodicalIF":2.8,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143705189","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}

{"title":"A validated liquid chromatography-tandem mass spectrometry assay for simultaneous quantitation of intact IGF-1 and IGF-2 in human serum","authors":"Hao-Long Zeng ,&nbsp;Jie Lu ,&nbsp;Huijun Li ,&nbsp;Liming Cheng","doi":"10.1016/j.jchromb.2025.124572","DOIUrl":"10.1016/j.jchromb.2025.124572","url":null,"abstract":"<div><div>The simultaneous quantification of insulin-like growth factor (IGF) 1 and 2 has demonstrated significant potential for routine diagnostic applications in growth disorders. However, a simplified and rapid assay still remains to be developed. In this study, we established a simple, cost-effective and antibody-free serological assay based on tandem mass spectrometry to simultaneously quantify intact IGF-1 and IGF-2 in human serum. The lower limits of quantification were estimated to be 18.16 ng/mL for IGF-1 and 23.71 ng/mL for IGF-2. The linearity correlation coefficients exceeded 0.99 for both analytes. No significant matrix effects or carryover were observed. Intra- and inter-assay precisions were both less than 15 %. Recoveries from the standard addition assay fell within the range of 80 % to 120 %. We subsequently assessed the serum levels of IGF-1 and IGF-2 in a population residing in Wuhan, China, and found IGF-1 levels exhibited an age-related increase, peaking between 10 and 20 years of age, whereas IGF-2 levels remained consistently elevated across all age groups, although a relative decrease was noted between the ages of 20 and 30 years. This assay provides a simple, rapid, and immunoaffinity-free approach, rendering it more suitable for clinical applications aimed at assessing serum IGF-1 and IGF-2 levels.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124572"},"PeriodicalIF":2.8,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143725936","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}

{"title":"Development and validation of an analytical method for metabolites of the plasticizer tri-(2-ethylhexyl) trimellitate (TOTM) in human urine by LC-MS/MS","authors":"Paulien Cleys ,&nbsp;Lucas Panneel ,&nbsp;Philippe G. Jorens ,&nbsp;Antonius Mulder ,&nbsp;Adrian Covaci","doi":"10.1016/j.jchromb.2025.124569","DOIUrl":"10.1016/j.jchromb.2025.124569","url":null,"abstract":"<div><div>Tri-(2-ethylhexyl) trimellitate (TEHTM or TOTM) is an alternative plasticizer used as replacement for conventional phthalates, such as bis-(2-ethyl-hexyl) phthalate (DEHP) in plastic materials. Migration studies show a lower leaching potential from the polymer matrix, compared to DEHP. Additionally, toxicological studies showed lower negative health effects of TOTM. A sensitive bioanalytical method is necessary to further enable the exposure assessment of TOTM. In this study, we developed a quantitative method based on SPE and LC-MS/MS for analysis of ten urinary metabolites of TOTM, such as 1,4-di-(2-ethylhexyl) trimellitate (1,4-DEHTM), 2,4-di-(2-ethylhexyl) trimellitate (2,4-DEHTM), 1-mono-(2-ethylhexyl) trimellitate (1-MEHTM), 2-mono-(2-ethylhexyl) trimellitate (2-MEHTM), 1-mono-(2-ethyl-5-carboxypentyl) trimellitate (5Cx-1-MEPTM), 2-mono-(2-ethyl-5-carboxypentyl) trimellitate (5Cx-2-MEPTM), 2-mono-(2-ethyl-5-hydroxyhexyl) trimellitate (5OH-2-MEHTM), and 1-mono-(2-ethyl-5-hydroxyhexyl) trimellitate (5OH-1-MEHTM), and 1-mono-(2-carboxymethylhexyl) trimellitate (2Cx-MMHTM). The method was validated according to the current ‘Bioanalytical Method Validation guidelines’ from European Medicines Agency (EMA). All validation criteria were successfully passed, except for 2Cx-MMHTM and 5Cx-2-MEPTM, which were discarded due to unsatisfactory validation results and low clinical relevance. Additionally, 1,4- and 2,4-DEHTM were successfully validated as a sum of two isomers. The validated method was applied to a pilot set of 30 urine samples from newborn patients. Significant higher urinary concentrations were found for 5Cx-1-MEPTM, 5OH-1-MEHTM, 5OH-2-MEHTM, 5oxo-1-MEHTM and 2-MEHTM in prematurely born babies from the neonatal intensive care unit (NICU) compared to term-born newborns. The current study expanded the range of analytes to eight TOTM metabolites which can be simultaneously analysed in urine samples. It also showed its clinical importance by analysing urine samples from newborns admitted to the NICU.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124569"},"PeriodicalIF":2.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143705188","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}

{"title":"Purification of chicoric acid from dandelion herbal pieces using macroporous resin combined with HSCCC and its antioxidant capacity in human keratinocytes","authors":"Ziwei Liang,&nbsp;Mengqi Wu,&nbsp;Jingying Xu,&nbsp;Yuxuan Liu,&nbsp;Qifeng Han,&nbsp;Xiang Yang,&nbsp;Wei Xia,&nbsp;Wenqing Zhang","doi":"10.1016/j.jchromb.2025.124567","DOIUrl":"10.1016/j.jchromb.2025.124567","url":null,"abstract":"<div><div>Dandelion (<em>Taraxacum mongolicum</em> Hand. - Mazz.) is a traditional Chinese medicinal herb rich in active ingredients such as flavonoids and polyphenols, which are used to treat swelling and inflammation-related diseases. In the present study, total flavonoids and polyphenols in dandelion herbal pieces (<em>Taraxaci Herba</em>) extracts were purified by HPD-500 resin column chromatography, and the total flavonoid purity in purified extracts E80% was 5.3 times that of before purification, reaching 43.98 %. The purified extracts E80% were analyzed by UHPLC–QTOF–MS and 8 phenolic acid compounds were identified, including caffeic acid, chlorogenic acid, and chicoric acid. Moreover, high-speed countercurrent chromatography method was established to separate chicoric acid from E80%. n-hexane-ethyl acetate-methanol-0.5 % acetic acid in water (2:7:2:7, <em>v</em>/v/<em>v</em>/v) was selected as the solvent system. Chicoric acid with a purity of 95.44 % was obtained with the flow rate of 2.0 mL/min, the speed of 850 r/min, the system temperature of 35 °C and sample loading of 100 mg. In vitro bioassay showed that chicoric acid presented impressive antioxidant activities in SDS-induced human keratinocytes by promoting cell proliferation, increasing superoxide dismutase (SOD) activity, reducing the reactive oxygen species (ROS) level, stabilizing the mitochondrial membrane potential, and inhibiting cell apoptosis. Collectively, these results lay a theoretical basis for the industrial separation of natural antioxidant compounds.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124567"},"PeriodicalIF":2.8,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687818","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}