Journal of Chromatography B最新文献

{"title":"Integrated untargeted and targeted metabolomics to reveal the mechanisms of herbal medicine HLXLD on Alzheimer's disease","authors":"Wei Zhou ,&nbsp;Ling Tian ,&nbsp;Xueqin Wang ,&nbsp;Guiyue Hou ,&nbsp;Jiao Yu ,&nbsp;Meige Chen ,&nbsp;Chenglong Xu ,&nbsp;Lijuan Xue ,&nbsp;Xiaojie Tan ,&nbsp;Ronghua Dai","doi":"10.1016/j.jchromb.2025.124746","DOIUrl":"10.1016/j.jchromb.2025.124746","url":null,"abstract":"<div><div>Huo-Luo-Xiao-Ling-Dan (HLXLD), a Chinese herbal medicine consisting of 11 herbs, has been shown to be effective in alleviating cognitive and memory impairment in Alzheimer's disease (AD). However, the underlying mechanisms require further investigation. This study aimed to clarify potential therapeutic mechanisms of HLXLD in the treatment of AD from a metabolic perspective. A rat model of AD induced by AlCl₃ and D-gal was established, the Morris water maze (MWM) test and hippocampal histopathology were used to evaluate the pharmacological effect of the HLXLD on AD. Subsequently, untargeted metabolomics of brain tissues samples was performed by UHPLC-Q-Exactive-Orbitrap-MS, followed by multivariate statistical analysis. Targeted metabolomics by UHPLC-QQQ-MS was further employed to validate and supplement the untargeted metabolomics finding, involving neurotransmitters, amino acids, arachidonic acid and lipids, to elucidate the relationship between disease, herbal medicine and metabolism pathway. The study found that HLXLD could relieve the progression of AD and regulate metabolic imbalances. The levels of 26 metabolites decreased and 6 increased in brain tissues including lysine, taurine, fumaric acid, prostaglandin E2, choline and so on, these altered metabolites were primarily associated with amino acid metabolism, TCA cycle, arachidonic acid metabolism and lipid metabolism. The targeted metabolomics results showed that compared with the model group, the levels of 8 neurotransmitters, 21 amino acids, 7 arachidonic acids and 16 lipids in brain tissue, 9 neurotransmitters, 20 amino acids, 6 arachidonic acids and 2 lipids in plasma were changed. In summary, HLXLD could improve the levels of endogenous metabolites by regulating multiple metabolic pathways and play a role in energy supply, anti-neuroinflammatory and neuroprotective effects in AD treatment.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1265 ","pages":"Article 124746"},"PeriodicalIF":2.8,"publicationDate":"2025-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144809548","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":"HILIC-FLD-based characterization of maternal plasma N-glycosylation profiles as novel diagnostic and predictive biomarkers for hemolytic disease of the fetus and newborn","authors":"Tingyi Li ,&nbsp;Zhou Qiu ,&nbsp;Yongcheng Li ,&nbsp;Yafei Yang ,&nbsp;Hou Qian ,&nbsp;Yujun Fan ,&nbsp;Yike Wu ,&nbsp;Wenlan Liu","doi":"10.1016/j.jchromb.2025.124744","DOIUrl":"10.1016/j.jchromb.2025.124744","url":null,"abstract":"<div><div>ABO incompatibility-induced hemolytic disease of the fetus and newborn (HDFN) lacks effective preventive measures, necessitating advanced strategies for early prenatal prediction. This study investigated whether pregnancy-associated maternal plasma N-glycome alterations enable accurate prediction and diagnosis of ABO-mediated HDFN. We conducted ultra high-resolution plasma N-glycomics using hydrophilic interaction liquid chromatography with fluorescence detection (HILIC-FLD) in a discovery cohort (40 HDFN cases vs. matched controls). Glycomic predictive models were developed through logistic regression of chromatographic glycan peaks (LglycoP) and feature-level analysis of glycosylation traits (LglycoF). Model performance was validated in an independent cohort (<em>n</em> = 42) using area under the receiver operator curve (AUC-ROC) with bootstrap resampling for stability assessment. Significant N-glycome dysregulation was identified in HDFN pregnancies, with 5 discriminative glycan peaks and altered glycosylation patterns featured by branching, trisialylation, di- and tri-galactosylation of glycans. The LglycoP model demonstrated robust capability of prediction and diagnosis of in both the discovery and validation cohort (AUC &gt; 0.80), respectively. This study establishes maternal plasma N-glycomic profiling as a precision tool for early prediction and diagnosis of ABO-HDFN, particularly for clinical false positive category of antibody titer test. Identification of unique plasma dysregulation of N-glycome features in ABO-HDFN pregnancies suggest potential mechanistic links between maternal glyco-inflammatory responses and fetal erythrocyte opsonization, offering new perspectives on ABO-HDFN pathogenesis.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1265 ","pages":"Article 124744"},"PeriodicalIF":2.8,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144773076","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":"HPLC analysis of gingerols and shogaols discriminated from ginger extracts with a diode array detector","authors":"Jinhui Liu ,&nbsp;Xiongjun Zuo ,&nbsp;Dan Wang ,&nbsp;Jin Lv ,&nbsp;Feng Long Gu","doi":"10.1016/j.jchromb.2025.124743","DOIUrl":"10.1016/j.jchromb.2025.124743","url":null,"abstract":"<div><div>A simple and inexpensive high performance liquid chromatography (HPLC) method for the analysis of gingerols and shogaols in ginger extracts discriminated with a diode array detector (DAD) has been developed. The ginger extracts were well separated with a linear gradient elution from mixed solvents of water and methanol to methanol within 30 min by using different C18 columns. The separated components were detected with a DAD. The gingerols and shogaols were discriminated from other components based on their same chromophore, which differs from other components. These components can be accurately and quantitatively analyzed contrast to the quality control method，INA method 114.000，which has severe separation and peak identification problems. Different samples prepared by solvent extraction and supercritical carbon dioxide extraction were analyzed by using our developed HPLC-DAD method. This method is quite promising for origin analysis of ginger rhizome slices with overlapped chromatograms of different detection wavelength.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124743"},"PeriodicalIF":2.8,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144712851","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":"Exploring the therapeutic potential of extractions of total flavonoids from Sarcandra Glabra in immune thrombocytopenia: A metabolomic and network pharmacology approach","authors":"Zhen Lu ,&nbsp;Yingjian Zeng ,&nbsp;Zhongkang Zhang ,&nbsp;Xiaonan Lu ,&nbsp;Guangbin Shang","doi":"10.1016/j.jchromb.2025.124742","DOIUrl":"10.1016/j.jchromb.2025.124742","url":null,"abstract":"<div><div><em>Sarcandra Glabra</em> is used in the treatment of Immune thrombocytopenia (ITP) clinically. Extractions of total flavonoids from <em>Sarcandra Glabra</em> (ETFSG) can improve platelet count in vivo and in vitro. However, the underlying mechanisms of ETFSG on ITP remain unclear. This study aims to investigate the mechanisms of ETFSG in the treatment of ITP via metabolomics and network pharmacology. Specifically, the number of differentiated megakaryocytes and platelet count in ITP model rats can be enhanced by the administration of ETFSG. Untargeted plasma metabolomics analysis revealed that ETFSG regulates the metabolic disorders of amino acids metabolism, arachidonic acid metabolism, pyrimidine metabolism, and citrate cycle metabolism pathways in the ITP animal model. Metabolomics-based network pharmacology analysis suggests that the PI3K-AKT signaling pathway may be involved in ETFSG's promotion of megakaryocyte differentiation. Western blot results indicate that ETFSG increases the expression of P-PI3K and P-AKT. In summary, these findings lay the groundwork for further exploration of the underlying mechanisms.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124742"},"PeriodicalIF":2.8,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144721761","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":"Targeted metabolomics analysis of 21 amino acids in serum of silicosis patients using HILIC-MS/MS","authors":"Huan Xu ,&nbsp;Mucen Xu ,&nbsp;Huiling Zhou ,&nbsp;Ao Pan ,&nbsp;Linyao Wang ,&nbsp;Jing Zhang ,&nbsp;Yongxin Li","doi":"10.1016/j.jchromb.2025.124736","DOIUrl":"10.1016/j.jchromb.2025.124736","url":null,"abstract":"<div><div>Silicosis is characterized by the formation of fibrotic lesions due to altered collagen synthesis, in which amino acids play a crucial role. However, targeted metabolomics studies of serum amino acids in silicosis patients remain limited. Herein, we developed a specific serum amino acid metabolomics assay for identifying differential amino acids as biomarkers, providing a sensitive and reliable basis for diagnosing silicosis. A hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the simultaneous quantification of 21 amino acids in serum was established by optimizing chromatographic and sample preparation conditions. The method validation results showed good linearity (<em>r</em> &gt; 0.99), recovery (78.3 %–128.4 %) and precision (1.27 %–13.01 %). The developed method was utilized to detect the levels of 21 amino acids in the serum of the silicosis patient group (SP; <em>n</em> = 104) and the healthy control group (HC; <em>n</em> = 118). The concentration of four amino acids (glutamate, arginine, aspartic acid and ornithine) was significantly different between the SP and the HC. Single diagnostic biomarkers exhibited area under the curve (AUC) values ranging from 0.712 to 0.892, while the glutamate-arginine combination achieved superior performance (AUC = 0.915, 86.1 % sensitivity, 90.0 % specificity). The aforementioned results indicate that four biomarkers were screened for the diagnosis of silicosis, with glutamate emerging as the most accurate single biomarker for predicting silicosis. The diagnostic performance was further enhanced by combining glutamate with arginine, which improved both sensitivity and specificity.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124736"},"PeriodicalIF":2.8,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144711118","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":"Quantification of dolutegravir, bictegravir, raltegravir and doravirine in human breastmilk with UPLC-MS/MS","authors":"van der Wekken - Pas L.C. ,&nbsp;van Ewijk - Beneken Kolmer E.W.J. ,&nbsp;L.H.M. te Brake,&nbsp;D.M. Burger","doi":"10.1016/j.jchromb.2025.124739","DOIUrl":"10.1016/j.jchromb.2025.124739","url":null,"abstract":"<div><div>With the use of newer and more robust antiretrovirals, the risk of viral transmission through breastfeeding has sharply diminished. This has led to a change in guideline recommendations. Currently, breastfeeding is regarded as an equipoise of formula, in case of a well-controlled HIV infection. However, it is not yet fully established whether infant exposure to antiretroviral drugs through breastmilk is a reason for concern. Only sparse and heterogenous data exist on concentrations of dolutegravir, raltegravir, bictegravir and doravirine in breastmilk. So further research is required and to accurately describe infant exposure of antiretrovirals through breastmilk, reproducible bioanalytical methods are needed.</div><div>Our existing UPLC-MS/MS method for several anti-HIV drugs in EDTA-plasma was modified for validation of dolutegravir, bictegravir, raltegravir and doravirine concentrations in human breastmilk. [¹³C,²H₅]-Dolutegravir, [¹³C,²H_2,¹⁵N]-bictegravir, [²H₆]-raltegravir and [¹³C₆]-doravirine were used as internal standard. The sample preparation involved protein precipitation and detection was performed with tandem mass spectrometry (MS/MS) in a total runtime of 10 min.</div><div>The assay was validated over the concentration range of 0.0100–10.0 mg/L for dolutegravir and doravirine, 0.00500–10.0 mg/L for raltegravir and 0.0200–20.0 mg/L for bictegravir. Within-run and between-run accuracy were within ±10 % of the nominal concentration and precision &lt;10 CV% for quality controls at high, medium and low concentrations, and ± 8.8 % and &lt; 14 CV%, respectively, at the lower limit of quantification for all analytes. Extraction recovery was 81 % and 79 % for dolutegravir and its internal standard, 83 % and 83 % for bictegravir and its internal standard, 89 % and 92 % for raltegravir and its internal standard and 104 % and 101 % for doravirine and its internal standard. Processed samples of dolutegravir, bictegravir, raltegravir and doravirine in breastmilk were 108 %, 95.5 %, 105 % and 102 % after 14 days at 4 °C.</div><div>The assay is currently being implemented successfully in pharmacokinetic studies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124739"},"PeriodicalIF":2.8,"publicationDate":"2025-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144694503","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 high-throughput LC-MS/MS method for simultaneous analysis of albendazole, albendazole sulfoxide and albendazole sulfone in human plasma","authors":"Nela Žideková ,&nbsp;Martin Kertys ,&nbsp;Juraj Mokrý ,&nbsp;Daniela Antolová ,&nbsp;Katarína Šimeková ,&nbsp;Róbert Rosoľanka","doi":"10.1016/j.jchromb.2025.124741","DOIUrl":"10.1016/j.jchromb.2025.124741","url":null,"abstract":"<div><div>Albendazole is a benzimidazole derivative with a broad spectrum of antihelmintic activity. Its primary metabolite, albendazole sulfoxide, is responsible for the treatment's efficacy. On the other hand, it contributes to side effects and toxicity. Therefore, their quantification in plasma may increase therapeutic success and reduce associated risks by maintaining values within the therapeutic range. The present study introduces an LC-MS/MS method for the simultaneous quantification of albendazole, albendazole sulfoxide, and albendazole sulfone in human plasma. A simple and sensitive LC-MS/MS method was developed to monitor the plasma levels of albendazole (0.25–200 ng/mL), albendazole sulfoxide (5–3500 ng/mL), and albendazole sulfone (0.5–500 ng/mL) with a coefficient of variation below 7 %. A one-step extraction procedure using an Ostro™ plate was applied, and the extracts were analysed by gradient elution followed by detection on a mass spectrometer in multiple reaction monitoring mode. The method offers several significant advantages, including a low sample volume (50 μL), a short run time (4 min), and is sufficiently linear to quantify both low and high concentrations of all analytes. The method was successfully validated according to the ICH guideline M10 on bioanalytical method validation, covering selectivity, linearity of the calibration curve, lower limit of quantification (LLOQ), accuracy, precision, dilution integrity, carry-over effect, matrix effects, extraction recovery, and stability. The fully developed and validated method was used to determine albendazole, albendazole sulfoxide, and albendazole sulfone in the plasma samples of ten patients, illustrating the monitoring of albendazole treatment in patients with alveolar echinococcosis.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124741"},"PeriodicalIF":2.8,"publicationDate":"2025-07-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144680000","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":"Magnetic solid-phase extraction combined with UPLC-MS/MS: A novel application for the analysis of alkylphenols and bisphenols in urine","authors":"Junlin Wang ,&nbsp;Nianhua Zhang ,&nbsp;Zhiyuan Wang,&nbsp;Peng Wang,&nbsp;Yiyao Cao","doi":"10.1016/j.jchromb.2025.124740","DOIUrl":"10.1016/j.jchromb.2025.124740","url":null,"abstract":"<div><div>A rapid, sensitive, simple, and accurate method was developed for the simultaneous determination of 3 alkylphenols (APs) and 11 bisphenols (BPs) in urine based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Following enzymatic hydrolysis of urine samples, corresponding isotope internal standards were added. Magnetic solid-phase extraction (M-SPE) technology was employed for separation and enrichment of analytes, followed by UPLC-MS/MS analysis. Results showed that within the defined concentration ranges, calibration curves for both APs and BPs exhibited excellent linearity, with all correlation coefficients (<em>r</em>) &gt;0.999. The method demonstrated excellent precision and accuracy, with intra- and inter-day coefficients of variation (CVs) &lt; 4.4 % and 4.9 %, respectively, and recoveries ranging from 80.8 % to 107.7 %. Limits of detection (LODs) of the 14 target analytes were 0.006 ng/mL–0.6 ng/mL, and limits of quantification (LOQs) were 0.02 ng/mL–2.0 ng/mL. When applied to quantify APs and BPs in urine samples, the method proved effective, demonstrating rapidity, sensitivity, simplicity, and accuracy, thus making it highly suitable for detecting APs and BPs in urine.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124740"},"PeriodicalIF":2.8,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144653686","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":"Exploring the potential of a quick and simultaneous DoE-based stability indicating novel RP-HPLC method for the estimation of capecitabine and curcumin in biodegradable nanoparticles and human plasma","authors":"Chethan Patil,&nbsp;Prasiddhi Naik,&nbsp;T. Mallamma,&nbsp;Prakash Goudanavar","doi":"10.1016/j.jchromb.2025.124731","DOIUrl":"10.1016/j.jchromb.2025.124731","url":null,"abstract":"<div><div>There is solid evidence that capecitabine (CAP) and curcumin (CUR) can alleviate symptoms of a wide range of illnesses. A fast and specific RP-HPLC method was designed and validated according to ICH requirements, employing an isosbestic point. This technology was chosen for its drug delivery application and synergistic effects of CAP and CUR as no HPLC approach has been documented to estimate and quantify both medicines simultaneously. The mobile phase used for the chromatographic separation was methanol: 0.1 % orthophosphoric acid, (80:20) and the conditions were 1 mL/min flow rate and 332 nm. Shim-pack Solar C18 (4.6 × 250 mm, 5 μm) HPLC column was used for this purpose. With a correlation coefficient value &gt;0.999, the created technique was determined to be linear across the concentration range of 0.25–16 μg/mL. The approach that was created was strong, accurate (with a recovery rate of 91.85–106.53 %), and precise (with a relative standard deviation of &lt;2.0 %). For CAP, the corresponding limits of detection and quantification were determined to be 0.02 and 0.08 μg/mL, whereas for CUR, they were 0.08 and 0.12 μg/mL, respectively. By ICH standards, the proposed approach was also tested in human plasma for validation. In addition, tests were conducted on the stress degradation and the established method's applicability to biodegradable nanoformulation. In the presence of degradation products, there was a good separation of drug peaks. The experimental model was shown to be significant (<em>P</em> &lt; 0.05) after a deliberate adjustment was assessed using analysis of variance. The Quality by Design methodology led to a more accurate procedure that produces consistent, reliable, high-quality data and accurately quantifies CAP and CUR in bulk and nanoparticulate systems.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124731"},"PeriodicalIF":2.8,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672588","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":"Optimization of natural deep eutectic solvent composition for ultrasound-assisted extraction of phenolic compound from propolis samples","authors":"Luciano A. de Albuquerque ,&nbsp;Marcos Levi C.M. dos Reis ,&nbsp;Débora de A. Santana ,&nbsp;Sérgio T. Oliva ,&nbsp;Maria Fernanda Silva ,&nbsp;Magdalena Espino ,&nbsp;Federico J.V. Gomez ,&nbsp;Fabio de S. Dias","doi":"10.1016/j.jchromb.2025.124737","DOIUrl":"10.1016/j.jchromb.2025.124737","url":null,"abstract":"<div><div>Propolis is a resinous substance collected by bees rich in bioactive phenolic compounds, which confer medicinal properties, making it highly commercialized. Thus, this study aimed to develop a green method of ultrasound-assisted extraction using a natural deep eutectic solvent (NADES) to determine bioactive phenolic compounds in propolis by HPLC-DAD. A mixture design was conducted to optimize the proportion of components in the NADES synthesis, evaluating the components lactic acid, glucose, and water. The best molar ratio was determined to be 2:1:30. To obtain the optimal experimental conditions for the phenolic compound extraction procedure using NADES, the factors NADES volume and sonication time were evaluated using a Doehlert matrix as the response surface methodology, and the optimal conditions were 3.5 mL and 21 min, respectively. Analytical curves were constructed based on the peak areas of standard solutions of vanillic acid, rutin, gallic acid, <em>trans</em>-cinnamic acid, catechin, syringic acid, d-hydroxybenzoic acid, <em>p</em>-coumaric acid, caffeic acid, chlorogenic acid, ferulic acid, and sinapic acid. The detection limits ranged from 0.09 to 0.4 mg L⁻¹. The method presented good precision and accuracy and was applied to determining 12 phenolic compounds in three propolis samples. Furthermore, the proposed method was evaluated as a green alternative for determining phenolic compounds in propolis samples using AGREE and AGREEprep metrics. The final result of these metrics confirms that this method is aligned with Green Analytical Chemistry (GAC) principles.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1264 ","pages":"Article 124737"},"PeriodicalIF":2.8,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144686128","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}