Journal of Chromatography B最新文献

{"title":"Intestinal microbiota-mediated serum pharmacochemistry reveals hepatoprotective metabolites of Platycodonis Radix against APAP-induced liver injury.","authors":"Yuan-Han Zhong, Xi-Wa Wu, Xin-Yu Zhang, Shou-Wen Zhang, Yan Feng, Xue-Mei Zhang, Bing-Bing Xu, Guo-Yue Zhong, Hui-Liang Huang, Jun-Wei He, Jin-Xiang Zeng, Jian Liang","doi":"10.1016/j.jchromb.2024.124395","DOIUrl":"https://doi.org/10.1016/j.jchromb.2024.124395","url":null,"abstract":"<p><p>The urgent need for new medications that regulate CYP2E1, CASP3, Nrf2, HO-1, TLR2, TLR4, STAT3, and NF-κB activities is paramount for the treatment of drug-induced liver injury (DILI), particularly from acetaminophen (APAP). Previous studies have suggested that platycosides of Platycodonis Radix exhibits hepatoprotective properties against APAP-induced liver injury (AILI), and their serum metabolites may be the effective agents. As the identify the serum metabolites of platycosides is a huge challenge, the mechanism whether platycosides exert effects through the serum metabolites regulating those targets still remain unclear. In this study, we propose a novel method termed intestinal microbiota-mediated serum pharmacochemistry (IMSP) to identify the serum metabolite profile of platycosides, using deglycosylated platycosides as template molecules. Our results identified a total of 44 prototype platycosides in the total platycosides fraction of Platycodonis Radix (PF). In rat serum, we identified 12 prototype platycosides and 45 metabolites derived from the 44 platycosides. Furthermore, our findings indicate that all 44 platycosides can enter the serum in the form of metabolites. The presence of these metabolites in serum is closely related to their oral bioavailability and the content of the prototypes. The in vivo animal experiments showed that the PF possessed significant anti-AILI effects and CYP2E1, CASP3, Nrf2, HO-1, TLR2, TLR4, STAT3, and NF-κB p65 regulation activities. And the in vitro cell experiments and molecular docking analyses further demonstrated that the hepatoprotective effects were mainly ascribed to the serum metabolites, which regulating targets of CYP2E1, CASP3, Nrf2, HO-1, TLR2, TLR4, STAT3, and NF-κB p65. Additionally, the activities of these metabolites are closely associated with their structures. In summary, the IMSP method significantly enhances the ability to identify platycoside metabolites in serum, reveals that all platycosides may contribute to anti-AILI activity through their metabolites, PF and some of these metabolites are promising candidate compounds for developing new medications with anti-AILI effects for the first time.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124395"},"PeriodicalIF":2.8,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142790811","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 crisugabalin (HSK16149) in biological matrix by LC-MS/MS method: An application to rat pharmacokinetic and tissue distribution studies.","authors":"Zeyu Wang, Pingming Tang, Caixia Dou, Jiale Shen, Ni Peng, Yao Li, Ju Wang, Xiaoyan Chen","doi":"10.1016/j.jchromb.2024.124396","DOIUrl":"https://doi.org/10.1016/j.jchromb.2024.124396","url":null,"abstract":"<p><p>Crisugabalin (HSK16149), a novel VGCC α2δ ligand, has been approved for the treatment of adult diabetic peripheral neuropathic pain (DPNP) and postherpetic neuralgia (PHN). In this study, an LC-MS/MS method was developed for the determination of crisugabalin in rat plasma and tissues homogenate. Samples were extracted by protein precipitation and separated on a Hypersil GOLD aQ column with methanol and 2 mM ammonium acetate in water containing 0.1 % formic acid as mobile phase. Crisugabalin and its internal standard HSK7891 were ionized by electrospray ionization source and detected by multiple reaction monitoring with transitions of m/z 210.9 → 134.4 and m/z 246.0 → 129.3. Over the range of 0.0100-10.0 μg/mL, the selectivity, linearity, precision and accuracy, matrix effect, stability, recovery and dilution integrity of crisugabalin were validated in rat plasma. Validation was also performed in rat liver homogenate at concentrations ranging from 0.0200-20.0 μg/g. The method was then successfully applied to determine the pharmacokinetics and tissue distribution of crisugabalin. In rats, orally administered crisugabalin was completely and rapidly absorbed with a peak time of about 0.57 h, and was mainly distributed to kidney, bladder and liver tissues. Crisugabalin exhibited linear pharmacokinetics over the oral dose range of 3-30 mg/kg.</p>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1251 ","pages":"124396"},"PeriodicalIF":2.8,"publicationDate":"2024-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142790812","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":"Identification and characterization of etomidate and metomidate metabolites in zebrafish, HLMs and S9 fraction by quadrupole-orbitrap LC-MS/MS for drug control","authors":"Shihao Zhong ,&nbsp;Shiyang Qin ,&nbsp;Yuanfeng Wang ,&nbsp;Huan Li ,&nbsp;Xiaoyi Wang ,&nbsp;Tianruomei Chai ,&nbsp;Jianghai Lu","doi":"10.1016/j.jchromb.2024.124374","DOIUrl":"10.1016/j.jchromb.2024.124374","url":null,"abstract":"<div><div>Etomidate is a common non-barbiturate anesthetic with psychoactivity, and metomidate, a structural modifier of etomidate, also has the same psychoactive effect, and the abuse of both has gradually intensified. In this study, etomidate, metomidate and their metabolic profiles with 4 days postfertilization (4dpf) zebrafish, 4 months postfertilization (4mpf) zebrafish, human liver microsomes (HLMs) and human liver S9 fraction were investigated using Liquid chromatography with high resolution mass spectrometry (LC-HRMS) for the first time. 14 etomidate metabolites and 11 metomidate metabolites were found, and the related metabolic pathways included monohydroxylation, dihydroxylation, dehydrogenation, N-dealkylation, O-dealkylation, oxidation, N-glucuronidation, O-glucuronidation and combination. Etomidate acid (E6 and M6) was considered a common biomarker for monitoring the abuse of etomidate and its analogues. Two characteristic metabolites (E4 and M4) could be used as biomarkers to monitor etomidate or metomidate abuse, respectively. Dealkylation, hydroxylation and glucuronidation were the main metabolic pathways of etomidate and metomidate. The differences in metabolic profiles of the three metabolic models were also compared for the first time. The results of this study can provide important reference for the detection of target compounds against the abuse of etomidate and metomidate, and the metabolic analysis of similar substances.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124374"},"PeriodicalIF":2.8,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142715218","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":"Spatial-resolved metabolome imaging of petals for Forsythia viridissima and Jasminum nudiflorum using online extraction (OLE) coupled to LC–Qtof-MS","authors":"Wenzheng Li ,&nbsp;Wei Li ,&nbsp;Hangyun He ,&nbsp;Maodong Wang ,&nbsp;Lijuan Wu ,&nbsp;Yang Yang ,&nbsp;Pengfei Tu ,&nbsp;Wenjing Liu ,&nbsp;Yuelin Song","doi":"10.1016/j.jchromb.2024.124385","DOIUrl":"10.1016/j.jchromb.2024.124385","url":null,"abstract":"<div><div>MS imaging (MSI) is a powerful technique for investigating the spatial distribution of metabolites in complex biological samples. However, due to the absence of liquid chromatography (LC) separation in routine MSI analysis, matrix effect is obvious and isomers identification remains challenging. To overcome these shortcomings of classical MSI tools (<em>e.g.</em>, DESI-MSI and MALDI-MSI) for isomer differentiation and insufficient datapoints for quantification, online extraction-liquid chromatogram–hybrid triple quadrupole-time-of-flight mass spectrometry (OLE-LC–Qtof-MS) platform has been developed for spatial metabolome. As a proof-of-concept, two species flowers namely <em>Forsythia viridissima</em> (FV) and <em>Jasminum nudiflorum</em> (JN) that bloom in early spring were collected, dried, and cut into small pieces (1.0 mm × 1.0 mm). All pieces successively underwent OLE-LC–Qtof-MS measurements. As a result, 46 and 41 metabolites were observed and identified from FV and JN petals, respectively. Particularly, each compound corresponded to a chromatographic peak and isomeric differentiation was achieved amongst a set of chlorogenic acid derivatives. The peak areas of high intensity metabolites were aligned and combined within either species. The datasets were individually converted into heatmaps for all compounds, 87 ones in total, and each grid of any heatmap was assigned to the original location in the petal. Then, the spatial-resolved distribution style of each compound crossing the petal was reflected by the re-organized heatmap bearing the petal shape. As expected, regio-specific occurrence and accumulation were observed for several compounds, particularly among the chlorogenic acid isomers. Above all, OLE-LC–Qtof-MS is an alternative tool for spatial-resolved metabolome attributing to the advantages of isomeric separation and reliable quantification.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124385"},"PeriodicalIF":2.8,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142737989","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 plasmid DNA using a novel two stage chromatography process","authors":"Minglei Yu ,&nbsp;Mengran Yu ,&nbsp;Feng Qian","doi":"10.1016/j.jchromb.2024.124381","DOIUrl":"10.1016/j.jchromb.2024.124381","url":null,"abstract":"<div><div>The chromatography process of large-scale plasmid purification with high efficiency and low cost has always been a major challenge. We established a two-step plasmid chromatography purification process combining multimodal and thiophilic chromatography with an overall chromatography yield of nearly 70%. Capto Core 700, a multimodal core–shell particle, was firstly used to remove the impurities from the crude lysate. The effects of different experimental conditions on chromatography recovery and impurity removal were screened. Compared to conventional size exclusion chromatography, the sample load and flow rate of this step were enhanced by 40-fold and 5-fold, respectively, while maintaining a 90% yield. For the thiophilic chromatography (Capto PlasmidSelect), the method of Design of Experiments (DoEs) was used to study the influence of parameters on the results. The effects of ammonium sulfate concentration, sodium chloride concentration and flowrate in the elution phase were studied and optimized with a central composite design model consisting of 17 experiments. The versatility of this process was demonstrated by successfully purifying three different lentiviral packaging plasmids (pLP1, pLP2 and pLP/VSVG) and the target plasmid containing green fluorescent protein (GFP). Purified plasmids consistently achieved a supercoiled purity of at least 90% with endotoxin levels below 5 EU/mg. Lentiviral vectors packaged using these plasmids exhibited high infectious titers of 1 × 10⁷ TU/mL, thereby verifying the process applicability for diverse plasmid purification requirements.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124381"},"PeriodicalIF":2.8,"publicationDate":"2024-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142745064","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":"Integrating lipidomics, 16S rRNA sequencing, and network pharmacology to explore the mechanism of Qikui granule in treating diabetic kidney disease mice","authors":"Qing You ,&nbsp;Yang Lin ,&nbsp;Jia-Hui Gong ,&nbsp;Wan-Yu Gui ,&nbsp;Qian-Hua Yan ,&nbsp;Jian-Dong Zou ,&nbsp;E-Hu Liu ,&nbsp;Chang-Yin Li","doi":"10.1016/j.jchromb.2024.124378","DOIUrl":"10.1016/j.jchromb.2024.124378","url":null,"abstract":"<div><div>Qikui granule (QKG), a hospital preparation of traditional Chinese medicine, has been widely used for diabetic kidney disease (DKD) in clinical practice. However, its holistic therapeutic effects and the underlying therapeutic mechanisms remain unclear. In the present study, the integrated analysis of network pharmacology, 16S rRNA sequencing, and non-targeted lipidomics was performed to explore the anti-DKD effects of QKG and the underlying mechanisms in <em>db/db</em> mouse DKD model. The results of the network pharmacology analysis identified the PI3K-AKT, EGFR, MAPK, JAK-STAT, FoxO, and AGE-RAGE signaling pathways as the potential molecular mechanisms responsible for the efficacy of QKG. Importantly, these signaling pathways were found to be closely related to lipid metabolism and gut microbiota. The therapeutic effectiveness of QKG against DKD was manifested by reducing body weight, alleviating oxidative stress, improving kidney function indicators, promoting the recovery of renal histopathological damage, and regulating the lipid metabolic profile of serum and kidney in <em>db/db</em> mice. A total of 26 lipid metabolites were identified as potential pharmacological biomarkers (PPBs) of QKG for the treatment of DKD, which were mainly involved in glycerophospholipid metabolism. Meanwhile, QKG could alleviate DKD-induced gut microbiota dysbiosis primarily by enriching <em>Candidatus_Arthromitus,</em> which showed a negative correlation with all 26 lipid PPBs as well as 5 biochemical parameters, including 2 oxidative stress factors and 3 kidney function indices. In conclusion, our findings suggest that QKG may upregulate the gut level of <em>Candidatus_Arthromitus</em> to suppress the abnormal activation of PI3K-AKT related signaling pathway, thereby reducing the levels of PC and LPC in the glycerophospholipid metabolism, to finally ameliorate the progression of DKD in <em>db/db</em> mice.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124378"},"PeriodicalIF":2.8,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694895","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":"Screening, fingerprinting, and identification of phenolic antioxidants in Persicaria chinensis (L.) H. Gross by liquid chromatography – electrochemical detection and liquid chromatography – tandem mass spectrometry","authors":"Xijin Yang ,&nbsp;Ying Li ,&nbsp;Qiju Shao ,&nbsp;Zhirong Li ,&nbsp;Zeli Chun ,&nbsp;Yan Wang ,&nbsp;Yaping Zhou ,&nbsp;Rongxiang Chen","doi":"10.1016/j.jchromb.2024.124387","DOIUrl":"10.1016/j.jchromb.2024.124387","url":null,"abstract":"<div><div><em>Persicaria chinensis</em> (L.) H. Gross (<em>P. chinensis</em>), also known as “Huotanmu” in Chinese, is a traditional ethnic herbal medicine and a functional food ingredient. In this study, liquid chromatography − electrochemical detection (LC-ECD) screening, liquid chromatography − tandem mass spectrometry (LC-MS/MS) identification, and multivariate statistical analysis were combined to clarify the antioxidant components in <em>P. chinensis</em>. The samples were extracted using 80 % aqueous methanol under ultrasound-assisted extraction and their in vitro antioxidant activities were evaluated. The fingerprint of <em>P. chinensis</em> was established using LC-ECD, while the identification of major chemical components was performed using LC-MS/MS through comparison of ion fragments. Subsequently, grey correlation analysis and partial least squares regression analysis were used to investigate the association of different compounds with antioxidant activity. <em>P. chinensis</em> from different sources showed excellent DPPH radical scavenging capacity, ABTS radical scavenging capacity, and ferric reducing antioxidant power. A total of 25 common peaks were obtained and identified in the LC-ECD fingerprint of <em>P. chinensis</em>, which were primarily comprised of tannins and flavonoids. The similarity between samples of different batches was ≥ 0.767, indicating relatively consistent quality. The comprehensive analysis results of grey relational analysis and partial least squares regression analysis indicated that tannins such as di-galloyl-HHDP-hexoside and davidiin were the major contributors to the antioxidant activity of <em>P. chinensis</em>. Therefore, LC-ECD can rapidly screen the potential phenolic antioxidants of <em>P. chinensis</em>, providing a basis for its product development, resource utilization, and quality control.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124387"},"PeriodicalIF":2.8,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142694914","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":"Application of 1-octanol in the extraction and GC-FID analysis of volatile organic compounds produced in biogas and biohydrogen processes","authors":"Weslei D. Pavini ,&nbsp;João V.S. Ferreira ,&nbsp;Danilo L. Flumignan ,&nbsp;Sandra I. Maintinguer ,&nbsp;José E. Oliveira ,&nbsp;Rodrigo Sequinel","doi":"10.1016/j.jchromb.2024.124376","DOIUrl":"10.1016/j.jchromb.2024.124376","url":null,"abstract":"<div><div>Information about the volatile organic compounds generated in biogas and hydrogen production bioreactors is essential to elucidate the metabolic routes and varying yields of CH₄ and H₂ processes. In this work, the determination of 12 compounds (acetone, methanol, ethanol, 1-propanol, 1-butanol, and acetic, propanoic, butyric, isovaleric, valeric, caproic, and lactic acids) was performed by gas chromatography, after a vortex-assisted liquid–liquid microextraction (VALLME) procedure using 1.2 mL of crude sample and 400 µL of 1-octanol. Optimization of the separation process was performed, considering the solvent viscosity. The analytical curves were validated using ANOVA, demonstrating satisfactory precision and accuracy. Selectivity was confirmed by GC–MS analysis, which allowed the detection of glycerol, 1,3-propanediol, and 1,3-butanediol in some samples. Methanol levels exceeded the upper limit of quantification, with acetic acid and ethanol being the predominant compounds in the analyzed reactors. An additional investigation was conducted to assess potential interferences for lactic acid. The developed method employs a biodegradable extraction solvent, without any need for a dispersing solvent, and involves a single chromatographic run, without any derivatization steps.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124376"},"PeriodicalIF":2.8,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142680347","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 application of a quantitative LC-MS/MS assay for the analysis of four dental local anesthetics in human plasma and breast milk","authors":"Shupeng Liu ,&nbsp;Xiaofei Wu ,&nbsp;Dan Liu ,&nbsp;Xiuqi Li ,&nbsp;Mengyang Yu ,&nbsp;Ming Lu ,&nbsp;Hongyun Wang","doi":"10.1016/j.jchromb.2024.124384","DOIUrl":"10.1016/j.jchromb.2024.124384","url":null,"abstract":"<div><div>Local anesthetics (LAs: articaine, lidocaine, bupivacaine, and mepivacaine) are essential for dental pain management. However, there are concerns that the lipophilic LAs could cross into breast milk causing toxicity to the infant. Our objective was to establish a multi-analyte LC-MS/MS method for the concurrent quantification of local anesthetics (LAs) in human plasma and breast milk, clarifying the transfer of LAs from plasma to breast milk, thereby offering crucial data for the safe assessment of LAs during the nursing period. Sample preparation for plasma and breast milk involved protein precipitation and a single dilution step. The chromatographic analysis was conducted using an ACQUITY UPLC BEH C18 column. Detection was facilitated by a triple quadrupole mass spectrometer employing positive electrospray ionization. The method demonstrated excellent linearity for articaine, mepivacaine, and bupivacaine across the range of 0.5 to 500 ng/mL, and for lidocaine from 1.0 to 1000 ng/mL in both plasma and breast milk, with correlation coefficients (R2) of 0.99 or higher. The method was effectively utilized in the analysis of samples from three nursing mothers who received dental treatment involving LAs. Articaine and lidocaine excretion in milk was confirmed, but it presented no significant safety risks to the breastfed infants.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124384"},"PeriodicalIF":2.8,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692292","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 PGC and Biphenyl stationary phases for the high throughput analysis of DNA epigenetic modifications by UHPLC-MS/MS","authors":"Quentin Vandoolaeghe ,&nbsp;Valérie Bouchart ,&nbsp;Yolaine Guérin ,&nbsp;Stéphanie Lagadu ,&nbsp;Claire Lopez-Piffet ,&nbsp;Raphaël Delépée","doi":"10.1016/j.jchromb.2024.124382","DOIUrl":"10.1016/j.jchromb.2024.124382","url":null,"abstract":"<div><div>Epigenetic alterations such as cytosine methylation, hydroxymethylation, formylation and carboxylation are well known modifications that are frequently associated with various disease such as cancer. These modifications are usually studied at the gene level to evaluate their impact on the expression of genes but there is a need for a whole genome quantification that can be more easily used as effect biomarkers. Here, we compare two high throughput methods for the UHPLC-MS/MS analysis of these four epigenetic markers in large cohort studies. The first method uses a porous graphitic carbon stationary phase modified by surface adsorption of triethylamine while the second method uses a biphenyl reversed phase. The two developed methods are then applied to 40 blood neutrophils DNA samples of former smokers and never-smokers from an agricultural occupational biobank. The results obtained shows no differences for the evaluation of 5-methylcytosine and 5-hydroxymethylcytosine with the two methods but highlight a diminution of both DNA methylation and hydroxymethylation in former smokers when compared to a never smoking population.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1250 ","pages":"Article 124382"},"PeriodicalIF":2.8,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142715216","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}