Journal of Chromatography B最新文献

{"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}

{"title":"Evaluation of ion source parameters and liquid chromatography methods for plasma untargeted metabolomics using orbitrap mass spectrometer","authors":"Hailemariam Abrha Assress ,&nbsp;Ahsan Hameed ,&nbsp;Lindsay M. Pack ,&nbsp;Mario G. Ferruzzi ,&nbsp;Renny S. Lan","doi":"10.1016/j.jchromb.2025.124564","DOIUrl":"10.1016/j.jchromb.2025.124564","url":null,"abstract":"<div><div>Although untargeted metabolomics holds promise for study of metabolites in human health and disease, robust method development and optimization are needed to reduce potential analytical biases and to ensure comprehensive, high-throughput results. In this study, the effect of mass spectrometer (MS) ion source parameters on the signal reproducibility and number of metabolite annotations during untargeted metabolomics is shown. Furthermore, different mobile phase gradients and columns (five reversed phase (RP)-C18 and two hydrophilic interaction liquid chromatography (HILIC) columns) were evaluated for untargeted metabolomics of blood plasma extracts. Positioning the electrospray needle at the farthest on the <em>Z</em>-direction and the closest tested position on the Y-direction with respect to the mass spectrometry inlet produced the best signal reproducibility and the greatest number of metabolite annotations. Moreover, optimal ion source conditions included a positive spray voltage between 2.5 and 3.5 kV, a negative spray voltage between 2.5 and 3.0 kV, vaporization and ion transfer tube (ITT) temperature between 250 and 350 °C, 30 to 50 arbitrary units of sheath gas, and at least 10 auxiliary gas units. Despite the differences in chromatographic characteristics, the different RP columns assessed showed comparable performance in terms of number of metabolites annotated. For HILIC columns, a zwitterionic column demonstrated better performance than an amide column. Finally, as compared with use of a RP column alone, use of both the optimal RP and HILIC approaches expanded metabolome coverage: the number of metabolites annotated increased by 60 %. This study highlights the significance of fine-tuning the MS ion source parameters and optimizing chromatographic conditions on metabolome coverage during untargeted metabolomics of plasma samples.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124564"},"PeriodicalIF":2.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143807424","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":"Rapid screening of cefotaxime resistance in Escherichia coli isolates by liquid chromatography with absorbance detection.","authors":"Yamima Tasnim,&nbsp;Hanin Diab,&nbsp;Sumon Sarkar,&nbsp;Md. Kaisar Rahman,&nbsp;Mariana Fernandez,&nbsp;Alexandra Calle,&nbsp;Jonathan E. Thompson,&nbsp;Babafela Awosile","doi":"10.1016/j.jchromb.2025.124565","DOIUrl":"10.1016/j.jchromb.2025.124565","url":null,"abstract":"<div><div>The clinical outcome of bacterial infection relies on proper and timely diagnosis of antibiotic susceptibility. We demonstrate that liquid chromatography (LC) with diode-array absorbance detection (DAD) accurately diagnoses antimicrobial resistance by measuring cefotaxime recovery following incubation with bacterial isolates either positive or negative for the cefotaximase (CTX-M) family of serine-β-lactamases. Reversed phase, high-performance liquid chromatography with absorbance detection was employed for cefotaxime analysis. A total of 43 <em>bla</em>_CTX-M<em>-</em>producing and 5 <em>bla</em>_CTX-M-negative <em>Escherichia coli</em> isolates were incubated with 0.5 mg/mL of cefotaxime at 37 °C for 1 and 2 h. Remarkably, after 2 h of incubation, the median ± median absolute deviation percentage of cefotaxime recovery was zero (0 %) for <em>bla</em>_CTX-M producing and cefotaxime-resistant <em>E. coli</em> isolates compared to the cefotaxime recovery in <em>bla</em>_CTX-M negative (61 ± 5.38 %) and cefotaxime-susceptible (70.68 ± 6.25 %) <em>E. coli</em> isolates. This result allows facile sorting of organism resistance status after only 1–2 h with near-perfect performance. The diagnostic sensitivity (Se) and specificity (Sp) of the chromatographic approach were comparable to widely used phenotypic (Epsilometer test, <em>E</em>-test) and genotypic assays (Whole Genome Sequencing, WGS), but chromatography reduces diagnostic time by &gt;10-fold. Additionally, the optical absorption measurement is fully compatible with microfluidic platforms, suggesting the development of low-cost, high-throughput sensors based on this measurement principle is possible. We conclude LC-DAD is suitable and reliable to determine the cefotaxime resistance status of <em>E. coli</em> isolates with a turn-around time of only 1–2 h.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124565"},"PeriodicalIF":2.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687819","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":"Novel GC MS/MS method for bioanalysis of pyrroloquinoline quinone, a potential cognitive enhancer in mice brains","authors":"Mai Mostafa ,&nbsp;Reham Abdel-Kader ,&nbsp;Rasha Hanafi","doi":"10.1016/j.jchromb.2025.124559","DOIUrl":"10.1016/j.jchromb.2025.124559","url":null,"abstract":"<div><div>Phenolic compounds have neuroprotective effect in diseases of cognitive impairment. Pyrroloquinoline quinone (PQQ), an aromatic water-soluble quinone enhances cognitive function <em>in-vivo</em> as previously demonstrated by our research group. In an attempt to comprehend the mechanism of action, development of a bioanalytical method for PQQ in brain matrix was essential to investigate blood-brain barrier (BBB) permeability for the drug and/or its metabolites. This study documents a novel fast GCMS/MS method for bioanalysis of PQQ in mice brains following a novel derivatization reaction of this drug. A simple extraction methodology using a single solvent highlights sustainability and greenness of our sample preparation protocol. Method validation and quantitative analysis of PQQ as an intact molecule in mice brain homogenates was done using novel qualifier and quantifier ions of the silylated drug for the first time. We report BBB permeation to PQQ in an induced neuroinflammation mouse model in addition to its sulfate metabolite following intraperitoneal injection. Interestingly, PQQ was detected in brains of control mice on standard diet containing soybeans. <em>In silico</em> prediction suggests the involvement of P-gp in active transport of PQQ across BBB where the drug appears to be is an excellent substrate and inhibitor. Pharmacokinetic analysis in brain revealed t_max as 2 h. Our optimized extraction method, as well as the GC–MS/MS method can be used to quantify levels of PQQ in various matrices opening the door to many other studies on this polyphenol. Moreover, we recommend the use of PQQ as a co-treatment in cognitive impairment diseases.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124559"},"PeriodicalIF":2.8,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143687874","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":"Leveraging HILIC/ERLIC separations for online nanoscale LC-MS/MS analysis of phosphopeptide isoforms from RNA polymerase II C-terminal domain","authors":"Scott B. Ficarro ,&nbsp;Deepash Kothiwal ,&nbsp;Hyun Jin Bae ,&nbsp;Isidoro Tavares ,&nbsp;Gabriela Giordano ,&nbsp;Stephen Buratowski ,&nbsp;Jarrod A. Marto","doi":"10.1016/j.jchromb.2025.124560","DOIUrl":"10.1016/j.jchromb.2025.124560","url":null,"abstract":"<div><div>The eukaryotic RNA polymerase II (Pol II) multi-protein complex transcribes mRNA and coordinates several steps of co-transcriptional mRNA processing and chromatin modification. The largest Pol II subunit, Rpb1, has a C-terminal domain (CTD) comprising dozens of repeated heptad sequences (Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7), each containing five phospho-accepting amino acids. The CTD heptads are dynamically phosphorylated, creating specific patterns correlated with steps of transcription initiation, elongation, and termination. This CTD phosphorylation ‘code’ choreographs dynamic recruitment of important co-regulatory proteins during gene transcription. Genetic tools were used to engineer protease cleavage sites across the CTD (msCTD), creating tryptic peptides with unique sequences amenable to mass spectrometry analysis. However, phosphorylation isoforms within each msCTD sequence are difficult to resolve by standard reversed phase chromatography typically used for LC-MS/MS applications. Here, we use a panel of synthetic CTD phosphopeptides to explore the potential of hydrophilic interaction and electrostatic repulsion hydrophilic interaction (HILIC and ERLIC) chromatography as alternatives to reversed phase separation for CTD phosphopeptide analysis. Our results demonstrate that ERLIC provides improved performance for separation of singly- and doubly-phosphorylated CTD peptides for sequence analysis by LC-MS/MS. Analysis of native yeast msCTD confirms that phosphorylation on Ser5 and Ser2 represents the major endogenous phosphoisoforms. We expect this methodology will be especially useful in the investigation of pathways where multiple protein phosphorylation events converge in close proximity.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1257 ","pages":"Article 124560"},"PeriodicalIF":2.8,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143725935","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":"LC-HRMS profiling of Dendrobium huoshanense aqueous extract and its therapeutic effects on nonalcoholic fatty liver disease in mice through the TLR2-NF-κB and AMPK-SREBP1-SIRT1 signaling pathways","authors":"Qiyan Lin ,&nbsp;Ke Huang ,&nbsp;Xiyu Ge ,&nbsp;Menghua Ma ,&nbsp;Wei Wang ,&nbsp;Li Yang ,&nbsp;Cunwu Chen ,&nbsp;Bangxing Han ,&nbsp;Dong Liu","doi":"10.1016/j.jchromb.2025.124563","DOIUrl":"10.1016/j.jchromb.2025.124563","url":null,"abstract":"<div><div><em>Dendrobium huoshanense</em> (DH) belongs to the <em>Dendrobium</em> genus of the Orchidaceae family and is a herbaceous plant that protects the liver and nourishes the Yin according to traditional Chinese Medicine (TCM) theory. This research aimed to determine the therapeutic effect and mechanisms of DH on a nonalcoholic fatty liver disease (NAFLD) mouse model and its chemical composition. For pharmacological research, the pathological damage and lipid accumulation in liver tissues were evaluated using HE and oil red staining, respectively. The differential proteins between the model and DHH groups were screened using 4D label-free quantitative proteomics, and the proteomic results were verified using Western blot. The potential mechanism was validated by metabolomic analysis. The main active ingredients in a DH aqueous extract were identified using UHPLC-Q Exactive HF HRMS. Pathological staining results showed that DH can reverse liver pathological damage and lipid accumulation in the NAFLD model. Quantitative proteomics revealed that the differential proteins were mainly associated with liver lipid deposition (LAL, AMPK, TM7SF2, SBCAD, and SIRT1), insulin resistance (GYS1, GYS2, PYGL, FoxO1, and PPAR-γ), and inflammation (TLR2 and MAPKAPK). Western blot verified the above-mentioned results. Metabolomic analysis also indicated that the DH aqueous extract ameliorated NAFLD in mice by affecting cholesterol metabolism and AMPK signaling pathway, proving its significant therapeutic effects on the NAFLD model. Sixty-five compounds were identified from DH aqueous extract by analyzing the precise molecular weight and MS/MS fragmentation pathway. The pharmacological mechanism of DH in treating NAFLD mainly involved the TLR2-NF-κB and AMPK-SREBP1-SIRT1 signaling pathways.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1256 ","pages":"Article 124563"},"PeriodicalIF":2.8,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143643178","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}